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Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
PowerPoint Lectures for
Biology, Seventh Edition
Neil Campbell and Jane Reece
Lectures by Chris Romero
Chapter 17
From Gene to Protein
Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
• Overview: The Flow of Genetic Information
• The information content of DNA
– Is in the form of specific sequences of
nucleotides along the DNA strands
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• The DNA inherited by an organism
– Leads to specific traits by dictating the
synthesis of proteins
• The process by which DNA directs protein
synthesis, gene expression
– Includes two stages, called transcription and
translation
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• The ribosome
– Is part of the cellular machinery for translation,
polypeptide synthesis
Figure 17.1
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• Concept 17.1: Genes specify proteins via
transcription and translation
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Evidence from the Study of Metabolic Defects
• In 1909, British physician Archibald Garrod
– Was the first to suggest that genes dictate
phenotypes through enzymes that catalyze
specific chemical reactions in the cell
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Nutritional Mutants in Neurospora: Scientific Inquiry
• Beadle and Tatum causes bread mold to
mutate with X-rays
– Creating mutants that could not survive on
minimal medium
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• Using genetic crosses
– They determined that their mutants fell into three
classes, each mutated in a different gene
Figure 17.2
Working with the mold Neurospora crassa, George Beadle and Edward Tatum had isolated mutants requiring
arginine in their growth medium and had shown genetically that these mutants fell into three classes, each
defective in a different gene. From other considerations, they suspected that the metabolic pathway of arginine
biosynthesis included the precursors ornithine and citrulline. Their most famous experiment, shown here,
tested both their one gene–one enzyme hypothesis and their postulated arginine pathway. In this experiment,
they grew their three classes of mutants under the four different conditions shown in the Results section below.
The wild-type strain required only the minimal medium for growth. The three classes of mutants had
different growth requirements
EXPERIMENT
RESULTS
Class I
Mutants
Class II
Mutants
Class III
MutantsWild type
Minimal
medium
(MM)
(control)
MM +
Ornithine
MM +
Citrulline
MM +
Arginine
(control)
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CONCLUSION From the growth patterns of the mutants, Beadle and Tatum deduced that each mutant was unable
to carry out one step in the pathway for synthesizing arginine, presumably because it lacked the
necessary enzyme. Because each of their mutants was mutated in a single gene, they concluded that
each mutated gene must normally dictate the production of one enzyme. Their results supported the
one gene–one enzyme hypothesis and also confirmed the arginine pathway.
(Notice that a mutant can grow only if supplied with a compound made after the defective step.)
Class I
Mutants
(mutation
in gene A)
Class II
Mutants
(mutation
in gene B)
Class III
Mutants
(mutation
in gene C)Wild type
Gene A
Gene B
Gene C
Precursor Precursor Precursor Precursor
Ornithine Ornithine Ornithine Ornithine
Citrulline Citrulline Citrulline Citrulline
Arginine Arginine Arginine Arginine
Enzyme
A
Enzyme
B
Enzyme
C
A A A
B B B
C C C
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• Beadle and Tatum developed the “one gene–
one enzyme hypothesis”
– Which states that the function of a gene is to
dictate the production of a specific enzyme
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The Products of Gene Expression: A Developing Story
• As researchers learned more about proteins
– The made minor revision to the one gene–one
enzyme hypothesis
• Genes code for polypeptide chains or for RNA
molecules
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Basic Principles of Transcription and Translation
• Transcription
– Is the synthesis of RNA under the direction of
DNA
– Produces messenger RNA (mRNA)
• Translation
– Is the actual synthesis of a polypeptide, which
occurs under the direction of mRNA
– Occurs on ribosomes
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• In prokaryotes
– Transcription and translation occur together
Figure 17.3a
Prokaryotic cell. In a cell lacking a nucleus, mRNA
produced by transcription is immediately translated
without additional processing.
(a)
TRANSLATION
TRANSCRIPTION
DNA
mRNA
Ribosome
Polypeptide
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• In eukaryotes
– RNA transcripts are modified before becoming
true mRNA
Figure 17.3b
Eukaryotic cell. The nucleus provides a separate
compartment for transcription. The original RNA
transcript, called pre-mRNA, is processed in various
ways before leaving the nucleus as mRNA.
(b)
TRANSCRIPTION
RNA PROCESSING
TRANSLATION
mRNA
DNA
Pre-mRNA
Polypeptide
Ribosome
Nuclear
envelope
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• Cells are governed by a cellular chain of
command
– DNA → RNA → protein
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The Genetic Code
• How many bases correspond to an amino
acid?
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Codons: Triplets of Bases
• Genetic information
– Is encoded as a sequence of nonoverlapping
base triplets, or codons
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• During transcription
– The gene determines the sequence of bases
along the length of an mRNA molecule
Figure 17.4
DNA
molecule
Gene 1
Gene 2
Gene 3
DNA strand
(template)
TRANSCRIPTION
mRNA
Protein
TRANSLATION
Amino acid
A C C A A A C C G A G T
U G G U U U G G C U C A
Trp Phe Gly Ser
Codon
3 5
35
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Cracking the Code
• A codon in messenger RNA
– Is either translated into an amino acid or serves as
a translational stop signal
Figure 17.5
Second mRNA base
U C A G
U
C
A
G
UUU
UUC
UUA
UUG
CUU
CUC
CUA
CUG
AUU
AUC
AUA
AUG
GUU
GUC
GUA
GUG
Met or
start
Phe
Leu
Leu
lle
Val
UCU
UCC
UCA
UCG
CCU
CCC
CCA
CCG
ACU
ACC
ACA
ACG
GCU
GCC
GCA
GCG
Ser
Pro
Thr
Ala
UAU
UAC
UGU
UGC
Tyr Cys
CAU
CAC
CAA
CAG
CGU
CGC
CGA
CGG
AAU
AAC
AAA
AAG
AGU
AGC
AGA
AGG
GAU
GAC
GAA
GAG
GGU
GGC
GGA
GGG
UGG
UAA
UAG Stop
Stop UGA Stop
Trp
His
Gln
Asn
Lys
Asp
Arg
Ser
Arg
Gly
U
C
A
G
U
C
A
G
U
C
A
G
U
C
A
G
FirstmRNAbase(5end)
ThirdmRNAbase(3end)
Glu
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• Codons must be read in the correct reading
frame
– For the specified polypeptide to be produced
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Evolution of the Genetic Code
• The genetic code is nearly universal
– Shared by organisms from the simplest
bacteria to the most complex animals
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• In laboratory experiments
– Genes can be transcribed and translated after
being transplanted from one species to
another
Figure 17.6
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• Concept 17.2: Transcription is the DNA-
directed synthesis of RNA: a closer look
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Molecular Components of Transcription
• RNA synthesis
– Is catalyzed by RNA polymerase, which pries
the DNA strands apart and hooks together the
RNA nucleotides
– Follows the same base-pairing rules as DNA,
except that in RNA, uracil substitutes for
thymine
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Synthesis of an RNA Transcript
• The stages of transcription are
– Initiation
– Elongation
– Termination
Figure 17.7
Promoter
Transcription unit
RNA polymerase
Start point
5
3
3
5
3
5
5
3
5
3
3
5
5
3
3
5
5
5
Rewound
RNA
RNA
transcript
3
3
Completed RNA
transcript
Unwound
DNA
RNA
transcript
Template strand of
DNA
DNA
1 Initiation. After RNA polymerase binds to
the promoter, the DNA strands unwind, and
the polymerase initiates RNA synthesis at the
start point on the template strand.
2 Elongation. The polymerase moves downstream, unwinding the
DNA and elongating the RNA transcript 5  3 . In the wake of
transcription, the DNA strands re-form a double helix.
3 Termination. Eventually, the RNA
transcript is released, and the
polymerase detaches from the DNA.
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Elongation
RNA
polymerase
Non-template
strand of DNA
RNA nucleotides
3 end
C A E G C A
A
U
T A G G T T
A
A
C
G
U
A
T
C
A
T C C A A
T
T
G
G
3
5
5
Newly made
RNA
Direction of transcription
(“downstream”) Template
strand of DNA
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RNA Polymerase Binding and Initiation of Transcription
• Promoters signal the initiation of RNA synthesis
• Transcription factors
– Help eukaryotic RNA polymerase recognize
promoter sequences
Figure 17.8Figure 17.8
TRANSCRIPTION
RNA PROCESSING
TRANSLATION
DNA
Pre-mRNA
mRNA
Ribosome
Polypeptide
T A T AAA A
ATAT T T T
TATA box Start point Template
DNA strand
5
3
3
5
Transcription
factors
5
3
3
5
Promoter
5
3
3
55
RNA polymerase II
Transcription factors
RNA transcript
Transcription initiation complex
Eukaryotic promoters1
Several transcription
factors
2
Additional transcription
factors
3
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Elongation of the RNA Strand
• As RNA polymerase moves along the DNA
– It continues to untwist the double helix,
exposing about 10 to 20 DNA bases at a time
for pairing with RNA nucleotides
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Termination of Transcription
• The mechanisms of termination
– Are different in prokaryotes and eukaryotes
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• Concept 17.3: Eukaryotic cells modify RNA
after transcription
• Enzymes in the eukaryotic nucleus
– Modify pre-mRNA in specific ways before the
genetic messages are dispatched to the
cytoplasm
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Alteration of mRNA Ends
• Each end of a pre-mRNA molecule is modified
in a particular way
– The 5′ end receives a modified nucleotide cap
– The 3′ end gets a poly-A tail
Figure 17.9
A modified guanine nucleotide
added to the 5 end
50 to 250 adenine nucleotides
added to the 3 end
Protein-coding segment Polyadenylation signal
Poly-A tail3 UTR
Stop codonStart codon
5 Cap 5 UTR
AAUAAA AAA…AAA
TRANSCRIPTION
RNA PROCESSING
DNA
Pre-mRNA
mRNA
TRANSLATION
Ribosome
Polypeptide
G P P P
5 3
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Split Genes and RNA Splicing
• RNA splicing
– Removes introns and joins exons
Figure 17.10
TRANSCRIPTION
RNA PROCESSING
DNA
Pre-mRNA
mRNA
TRANSLATION
Ribosome
Polypeptide
5 Cap
Exon Intron
1
5
30 31
Exon Intron
104 105 146
Exon 3
Poly-A tail
Poly-A tail
Introns cut out and
exons spliced together
Coding
segment
5 Cap
1 146
3 UTR3 UTR
Pre-mRNA
mRNA
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• Is carried out by spliceosomes in some cases
Figure 17.11
RNA transcript (pre-mRNA)
Exon 1 Intron Exon 2
Other proteins
Protein
snRNA
snRNPs
Spliceosome
Spliceosome
components
Cut-out
intron
mRNA
Exon 1 Exon 2
5
5
5
1
2
3
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Ribozymes
• Ribozymes
– Are catalytic RNA molecules that function as
enzymes and can splice RNA
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The Functional and Evolutionary Importance of Introns
• The presence of introns
– Allows for alternative RNA splicing
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• Proteins often have a modular architecture
– Consisting of discrete structural and functional
regions called domains
• In many cases
– Different exons code for the different domains in a
protein
Figure 17.12
Gene
DNA
Exon 1 Intron Exon 2 Intron Exon 3
Transcription
RNA processing
Translation
Domain 3
Domain 1
Domain 2
Polypeptide
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• Concept 17.4: Translation is the RNA-directed
synthesis of a polypeptide: a closer look
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Molecular Components of Translation
• A cell translates an mRNA message into
protein
– With the help of transfer RNA (tRNA)
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• Translation: the basic concept
Figure 17.13
TRANSCRIPTION
TRANSLATION
DNA
mRNA
Ribosome
Polypeptide
Polypeptide
Amino
acids
tRNA with
amino acid
attachedRibosome
tRNA
Anticodon
mRNA
Trp
Phe Gly
A
G C
A A A
C
C
G
U G G U U U G G C
Codons5 3
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• Molecules of tRNA are not all identical
– Each carries a specific amino acid on one end
– Each has an anticodon on the other end
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The Structure and Function of Transfer RNA
A
C
C
• A tRNA molecule
– Consists of a single RNA strand that is only
about 80 nucleotides long
– Is roughly L-shaped
Figure 17.14a
Two-dimensional structure. The four base-paired regions and
three loops are characteristic of all tRNAs, as is the base sequence
of the amino acid attachment site at the 3 end. The anticodon triplet
is unique to each tRNA type. (The asterisks mark bases that have
been chemically modified, a characteristic of tRNA.)
(a)
3
C
C
A
C
G
C
U
U
A
A
GACAC
CU
*
G
C
* *
G U G U
*CU
* G AG
G
U
*
*A
*
A
A G
U
C
A
G
A
C
C
*
C G A G
A G G
G
*
*
GA
CUC*A
U
U
U
A
G
G
C
G
5
Amino acid
attachment site
Hydrogen
bonds
Anticodon
A
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Figure 17.14b
(b) Three-dimensional structure
Symbol used
in this book
Amino acid
attachment site
Hydrogen
bonds
Anticodon
Anticodon
A AG
5
3
3 5
(c)
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• A specific enzyme called an aminoacyl-tRNA
synthetase
– Joins each amino acid to the correct tRNA
Figure 17.15
Amino acid
ATP
Adenosine
Pyrophosphate
Adenosine
Adenosine
Phosphates
tRNA
P P P
P
P Pi
Pi
Pi
P
AMP
Aminoacyl tRNA
(an “activated
amino acid”)
Aminoacyl-tRNA
synthetase (enzyme)
Active site binds the
amino acid and ATP.
1
ATP loses two P groups
and joins amino acid as AMP.
2
3 Appropriate
tRNA covalently
Bonds to amino
Acid, displacing
AMP.
Activated amino acid
is released by the enzyme.
4
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Ribosomes
• Ribosomes
– Facilitate the specific coupling of tRNA
anticodons with mRNA codons during protein
synthesis
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• The ribosomal subunits
– Are constructed of proteins and RNA
molecules named ribosomal RNA or rRNA
Figure 17.16a
TRANSCRIPTION
TRANSLATION
DNA
mRNA
Ribosome
Polypeptide
Exit tunnel
Growing
polypeptide
tRNA
molecules
E
P A
Large
subunit
Small
subunit
mRNA
Computer model of functioning ribosome. This is a model of a bacterial
ribosome, showing its overall shape. The eukaryotic ribosome is roughly
similar. A ribosomal subunit is an aggregate of ribosomal RNA molecules
and proteins.
(a)
5
3
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• The ribosome has three binding sites for tRNA
– The P site
– The A site
– The E site
Figure 17.16b
E P A
P site (Peptidyl-tRNA
binding site)
E site
(Exit site)
mRNA
binding site
A site (Aminoacyl-
tRNA binding site)
Large
subunit
Small
subunit
Schematic model showing binding sites. A ribosome has an mRNA
binding site and three tRNA binding sites, known as the A, P, and E
sites. This schematic ribosome will appear in later diagrams.
(b)
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Figure 17.16c
Amino end Growing polypeptide
Next amino acid
to be added to
polypeptide chain
tRNA
mRNA
Codons
3
5
Schematic model with mRNA and tRNA. A tRNA fits into a binding site when its anticodon
base-pairs with an mRNA codon. The P site holds the tRNA attached to the growing
polypeptide. The A site holds the tRNA carrying the next amino acid to be added to the
polypeptide chain. Discharged tRNA leaves via the E site.
(c)
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Building a Polypeptide
• We can divide translation into three stages
– Initiation
– Elongation
– Termination
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Ribosome Association and Initiation of Translation
• The initiation stage of translation
– Brings together mRNA, tRNA bearing the first
amino acid of the polypeptide, and two
subunits of a ribosome
Large
ribosomal
subunit
The arrival of a large ribosomal subunit completes
the initiation complex. Proteins called initiation
factors (not shown) are required to bring all the
translation components together. GTP provides
the energy for the assembly. The initiator tRNA is
in the P site; the A site is available to the tRNA
bearing the next amino acid.
2
Initiator tRNA
mRNA
mRNA binding site Small
ribosomal
subunit
Translation initiation complex
P site
GDPGTP
Start codon
A small ribosomal subunit binds to a molecule of
mRNA. In a prokaryotic cell, the mRNA binding site
on this subunit recognizes a specific nucleotide
sequence on the mRNA just upstream of the start
codon. An initiator tRNA, with the anticodon UAC,
base-pairs with the start codon, AUG. This tRNA
carries the amino acid methionine (Met).
1
Met
Met
U A C
A U G
E A
3
5
5
3
35 35
Figure 17.17
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Elongation of the Polypeptide Chain
• In the elongation stage of translation
– Amino acids are added one by one to the
preceding amino acid
Figure 17.18
Amino end
of polypeptide
mRNA
Ribosome ready for
next aminoacyl tRNA
E
P A
E
P A
E
P A
E
P A
GDP
GTP
GTP
GDP
2
2
site site5
3
TRANSCRIPTION
TRANSLATION
DNA
mRNA
Ribosome
Polypeptide
Codon recognition. The anticodon
of an incoming aminoacyl tRNA
base-pairs with the complementary
mRNA codon in the A site. Hydrolysis
of GTP increases the accuracy and
efficiency of this step.
1
Peptide bond formation. An
rRNA molecule of the large
subunit catalyzes the formation
of a peptide bond between the
new amino acid in the A site and
the carboxyl end of the growing
polypeptide in the P site. This step
attaches the polypeptide to the
tRNA in the A site.
2
Translocation. The ribosome
translocates the tRNA in the A
site to the P site. The empty tRNA
in the P site is moved to the E site,
where it is released. The mRNA
moves along with its bound tRNAs,
bringing the next codon to be
translated into the A site.
3
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Termination of Translation
• The final stage of translation is termination
– When the ribosome reaches a stop codon in
the mRNA
Figure 17.19
Release
factor
Free
polypeptide
Stop codon
(UAG, UAA, or UGA)
5
3 3
5
3
5
When a ribosome reaches a stop
codon on mRNA, the A site of the
ribosome accepts a protein called
a release factor instead of tRNA.
1 The release factor hydrolyzes
the bond between the tRNA in
the P site and the last amino
acid of the polypeptide chain.
The polypeptide is thus freed
from the ribosome.
2 3 The two ribosomal subunits
and the other components of
the assembly dissociate.
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Polyribosomes
• A number of ribosomes can translate a single
mRNA molecule simultaneously
– Forming a polyribosome
Figure 17.20a, b
Growing
polypeptides
Completed
polypeptide
Incoming
ribosomal
subunits
Start of
mRNA
(5 end)
End of
mRNA
(3 end)
Polyribosome
An mRNA molecule is generally translated simultaneously
by several ribosomes in clusters called polyribosomes.
(a)
Ribosomes
mRNA
This micrograph shows a large polyribosome in a prokaryotic
cell (TEM).
0.1 µm
(b)
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Completing and Targeting the Functional Protein
• Polypeptide chains
– Undergo modifications after the translation
process
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Protein Folding and Post-Translational Modifications
• After translation
– Proteins may be modified in ways that affect
their three-dimensional shape
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Targeting Polypeptides to Specific Locations
• Two populations of ribosomes are evident in
cells
– Free and bound
• Free ribosomes in the cytosol
– Initiate the synthesis of all proteins
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• Proteins destined for the endomembrane
system or for secretion
– Must be transported into the ER
– Have signal peptides to which a signal-
recognition particle (SRP) binds, enabling the
translation ribosome to bind to the ER
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Figure 17.21
Ribosome
mRNA
Signal
peptide
Signal-
recognition
particle
(SRP) SRP
receptor
protein
Translocation
complex
CYTOSOL
Signal
peptide
removed
ER
membrane
Protein
ERLUMEN
• The signal mechanism for targeting proteins to
the ER
Polypeptide
synthesis begins
on a free
ribosome in
the cytosol.
1 An SRP binds
to the signal
peptide, halting
synthesis
momentarily.
2 The SRP binds to a
receptor protein in the ER
membrane. This receptor
is part of a protein complex
(a translocation complex)
that has a membrane pore
and a signal-cleaving enzyme.
3 The SRP leaves, and
the polypeptide resumes
growing, meanwhile
translocating across the
membrane. (The signal
peptide stays attached
to the membrane.)
4 The signal-
cleaving
enzyme
cuts off the
signal peptide.
5 The rest of
the completed
polypeptide leaves
the ribosome and
folds into its final
conformation.
6
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• Concept 17.5: RNA plays multiple roles in the
cell: a review
• RNA
– Can hydrogen-bond to other nucleic acid
molecules
– Can assume a specific three-dimensional
shape
– Has functional groups that allow it to act as a
catalyst
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• Types of RNA in a Eukaryotic Cell
Table 17.1
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• Concept 17.6: Comparing gene expression in
prokaryotes and eukaryotes reveals key differences
• Prokaryotic cells lack a nuclear envelope
– Allowing translation to begin while transcription is
still in progress
Figure 17.22
DNA
Polyribosome
mRNA
Direction of
transcription
0.25 mRNA
polymerase
Polyribosome
Ribosome
DNA
mRNA (5 end)
RNA polymerase
Polypeptide
(amino end)
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• In a eukaryotic cell
– The nuclear envelope separates transcription
from translation
– Extensive RNA processing occurs in the
nucleus
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• Concept 17.7: Point mutations can affect
protein structure and function
• Mutations
– Are changes in the genetic material of a cell
• Point mutations
– Are changes in just one base pair of a gene
Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
• The change of a single nucleotide in the DNA’s
template strand
– Leads to the production of an abnormal protein
Figure 17.23
In the DNA, the
mutant template
strand has an A where
the wild-type template
has a T.
The mutant mRNA has
a U instead of an A in
one codon.
The mutant (sickle-cell)
hemoglobin has a valine
(Val) instead of a glutamic
acid (Glu).
Mutant hemoglobin DNAWild-type hemoglobin DNA
mRNA mRNA
Normal hemoglobin Sickle-cell hemoglobin
Glu Val
C T T C A T
G A A G U A
3 5 3 5
5 35 3
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Types of Point Mutations
• Point mutations within a gene can be divided
into two general categories
– Base-pair substitutions
– Base-pair insertions or deletions
Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
Substitutions
• A base-pair substitution
– Is the replacement of one nucleotide and its
partner with another pair of nucleotides
– Can cause missense or nonsense
Figure 17.24
Wild type
A U G A A G U U U G G C U A A
mRNA
5
Protein Met Lys Phe Gly Stop
Carboxyl end
Amino end
3
A U G A A G U U U G G U U A A
Met Lys Phe Gly
Base-pair substitution
No effect on amino acid sequence
U instead of C
Stop
A U G A A G U U U A G U U A A
Met Lys Phe Ser Stop
A U G U A G U U U G G C U A A
Met Stop
Missense A instead of G
Nonsense
U instead of A
Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
Insertions and Deletions
• Insertions and deletions
– Are additions or losses of nucleotide pairs in a
gene
– May produce frameshift mutations
Figure 17.25
mRNA
Protein
Wild type
A U G A A G U U U G G C U A A
5
Met Lys Phe Gly
Amino end Carboxyl end
Stop
Base-pair insertion or deletion
Frameshift causing immediate nonsense
A U G U A A G U U U G G C U A
A U G A A G U U G G C U A A
A U G U U U G G C U A A
Met Stop
U
Met Lys Leu Ala
Met Phe Gly
Stop
MissingA A G
Missing
Extra U
Frameshift causing
extensive missense
Insertion or deletion of 3 nucleotides:
no frameshift but extra or missing amino acid
3
Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
Mutagens
• Spontaneous mutations
– Can occur during DNA replication,
recombination, or repair
Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
• Mutagens
– Are physical or chemical agents that can
cause mutations
Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
What is a gene? revisiting the question
• A gene
– Is a region of DNA whose final product is either
a polypeptide or an RNA molecule
Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
• A summary of transcription and translation in a
eukaryotic cell
Figure 17.26
TRANSCRIPTION
RNA is transcribed
from a DNA template.
DNA
RNA
polymerase
RNA
transcript
RNA PROCESSING
In eukaryotes, the
RNA transcript (pre-
mRNA) is spliced and
modified to produce
mRNA, which moves
from the nucleus to the
cytoplasm.
Exon
Poly-A
RNA transcript
(pre-mRNA)
Intron
NUCLEUS
Cap
FORMATION OF
INITIATION COMPLEX
After leaving the
nucleus, mRNA attaches
to the ribosome.
CYTOPLASM
mRNA
Poly-A
Growing
polypeptide
Ribosomal
subunits
Cap
Aminoacyl-tRNA
synthetase
Amino
acid
tRNA
AMINO ACID ACTIVATION
Each amino acid
attaches to its proper tRNA
with the help of a specific
enzyme and ATP.
Activated
amino acid
TRANSLATION
A succession of tRNAs
add their amino acids to
the polypeptide chain
as the mRNA is moved
through the ribosome
one codon at a time.
(When completed, the
polypeptide is released
from the ribosome.)
Anticodon
A CC
A A A
UG GUU UA U G
UACE A
Ribosome
1
Poly-A
5
5
3
Codon
2
3 4
5

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Gene to Protein Flow in Cells

  • 1. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings PowerPoint Lectures for Biology, Seventh Edition Neil Campbell and Jane Reece Lectures by Chris Romero Chapter 17 From Gene to Protein
  • 2. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Overview: The Flow of Genetic Information • The information content of DNA – Is in the form of specific sequences of nucleotides along the DNA strands
  • 3. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • The DNA inherited by an organism – Leads to specific traits by dictating the synthesis of proteins • The process by which DNA directs protein synthesis, gene expression – Includes two stages, called transcription and translation
  • 4. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • The ribosome – Is part of the cellular machinery for translation, polypeptide synthesis Figure 17.1
  • 5. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Concept 17.1: Genes specify proteins via transcription and translation
  • 6. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Evidence from the Study of Metabolic Defects • In 1909, British physician Archibald Garrod – Was the first to suggest that genes dictate phenotypes through enzymes that catalyze specific chemical reactions in the cell
  • 7. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Nutritional Mutants in Neurospora: Scientific Inquiry • Beadle and Tatum causes bread mold to mutate with X-rays – Creating mutants that could not survive on minimal medium
  • 8. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Using genetic crosses – They determined that their mutants fell into three classes, each mutated in a different gene Figure 17.2 Working with the mold Neurospora crassa, George Beadle and Edward Tatum had isolated mutants requiring arginine in their growth medium and had shown genetically that these mutants fell into three classes, each defective in a different gene. From other considerations, they suspected that the metabolic pathway of arginine biosynthesis included the precursors ornithine and citrulline. Their most famous experiment, shown here, tested both their one gene–one enzyme hypothesis and their postulated arginine pathway. In this experiment, they grew their three classes of mutants under the four different conditions shown in the Results section below. The wild-type strain required only the minimal medium for growth. The three classes of mutants had different growth requirements EXPERIMENT RESULTS Class I Mutants Class II Mutants Class III MutantsWild type Minimal medium (MM) (control) MM + Ornithine MM + Citrulline MM + Arginine (control)
  • 9. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings CONCLUSION From the growth patterns of the mutants, Beadle and Tatum deduced that each mutant was unable to carry out one step in the pathway for synthesizing arginine, presumably because it lacked the necessary enzyme. Because each of their mutants was mutated in a single gene, they concluded that each mutated gene must normally dictate the production of one enzyme. Their results supported the one gene–one enzyme hypothesis and also confirmed the arginine pathway. (Notice that a mutant can grow only if supplied with a compound made after the defective step.) Class I Mutants (mutation in gene A) Class II Mutants (mutation in gene B) Class III Mutants (mutation in gene C)Wild type Gene A Gene B Gene C Precursor Precursor Precursor Precursor Ornithine Ornithine Ornithine Ornithine Citrulline Citrulline Citrulline Citrulline Arginine Arginine Arginine Arginine Enzyme A Enzyme B Enzyme C A A A B B B C C C
  • 10. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Beadle and Tatum developed the “one gene– one enzyme hypothesis” – Which states that the function of a gene is to dictate the production of a specific enzyme
  • 11. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The Products of Gene Expression: A Developing Story • As researchers learned more about proteins – The made minor revision to the one gene–one enzyme hypothesis • Genes code for polypeptide chains or for RNA molecules
  • 12. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Basic Principles of Transcription and Translation • Transcription – Is the synthesis of RNA under the direction of DNA – Produces messenger RNA (mRNA) • Translation – Is the actual synthesis of a polypeptide, which occurs under the direction of mRNA – Occurs on ribosomes
  • 13. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • In prokaryotes – Transcription and translation occur together Figure 17.3a Prokaryotic cell. In a cell lacking a nucleus, mRNA produced by transcription is immediately translated without additional processing. (a) TRANSLATION TRANSCRIPTION DNA mRNA Ribosome Polypeptide
  • 14. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • In eukaryotes – RNA transcripts are modified before becoming true mRNA Figure 17.3b Eukaryotic cell. The nucleus provides a separate compartment for transcription. The original RNA transcript, called pre-mRNA, is processed in various ways before leaving the nucleus as mRNA. (b) TRANSCRIPTION RNA PROCESSING TRANSLATION mRNA DNA Pre-mRNA Polypeptide Ribosome Nuclear envelope
  • 15. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Cells are governed by a cellular chain of command – DNA → RNA → protein
  • 16. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The Genetic Code • How many bases correspond to an amino acid?
  • 17. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Codons: Triplets of Bases • Genetic information – Is encoded as a sequence of nonoverlapping base triplets, or codons
  • 18. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • During transcription – The gene determines the sequence of bases along the length of an mRNA molecule Figure 17.4 DNA molecule Gene 1 Gene 2 Gene 3 DNA strand (template) TRANSCRIPTION mRNA Protein TRANSLATION Amino acid A C C A A A C C G A G T U G G U U U G G C U C A Trp Phe Gly Ser Codon 3 5 35
  • 19. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Cracking the Code • A codon in messenger RNA – Is either translated into an amino acid or serves as a translational stop signal Figure 17.5 Second mRNA base U C A G U C A G UUU UUC UUA UUG CUU CUC CUA CUG AUU AUC AUA AUG GUU GUC GUA GUG Met or start Phe Leu Leu lle Val UCU UCC UCA UCG CCU CCC CCA CCG ACU ACC ACA ACG GCU GCC GCA GCG Ser Pro Thr Ala UAU UAC UGU UGC Tyr Cys CAU CAC CAA CAG CGU CGC CGA CGG AAU AAC AAA AAG AGU AGC AGA AGG GAU GAC GAA GAG GGU GGC GGA GGG UGG UAA UAG Stop Stop UGA Stop Trp His Gln Asn Lys Asp Arg Ser Arg Gly U C A G U C A G U C A G U C A G FirstmRNAbase(5end) ThirdmRNAbase(3end) Glu
  • 20. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Codons must be read in the correct reading frame – For the specified polypeptide to be produced
  • 21. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Evolution of the Genetic Code • The genetic code is nearly universal – Shared by organisms from the simplest bacteria to the most complex animals
  • 22. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • In laboratory experiments – Genes can be transcribed and translated after being transplanted from one species to another Figure 17.6
  • 23. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Concept 17.2: Transcription is the DNA- directed synthesis of RNA: a closer look
  • 24. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Molecular Components of Transcription • RNA synthesis – Is catalyzed by RNA polymerase, which pries the DNA strands apart and hooks together the RNA nucleotides – Follows the same base-pairing rules as DNA, except that in RNA, uracil substitutes for thymine
  • 25. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Synthesis of an RNA Transcript • The stages of transcription are – Initiation – Elongation – Termination Figure 17.7 Promoter Transcription unit RNA polymerase Start point 5 3 3 5 3 5 5 3 5 3 3 5 5 3 3 5 5 5 Rewound RNA RNA transcript 3 3 Completed RNA transcript Unwound DNA RNA transcript Template strand of DNA DNA 1 Initiation. After RNA polymerase binds to the promoter, the DNA strands unwind, and the polymerase initiates RNA synthesis at the start point on the template strand. 2 Elongation. The polymerase moves downstream, unwinding the DNA and elongating the RNA transcript 5  3 . In the wake of transcription, the DNA strands re-form a double helix. 3 Termination. Eventually, the RNA transcript is released, and the polymerase detaches from the DNA.
  • 26. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Elongation RNA polymerase Non-template strand of DNA RNA nucleotides 3 end C A E G C A A U T A G G T T A A C G U A T C A T C C A A T T G G 3 5 5 Newly made RNA Direction of transcription (“downstream”) Template strand of DNA
  • 27. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings RNA Polymerase Binding and Initiation of Transcription • Promoters signal the initiation of RNA synthesis • Transcription factors – Help eukaryotic RNA polymerase recognize promoter sequences Figure 17.8Figure 17.8 TRANSCRIPTION RNA PROCESSING TRANSLATION DNA Pre-mRNA mRNA Ribosome Polypeptide T A T AAA A ATAT T T T TATA box Start point Template DNA strand 5 3 3 5 Transcription factors 5 3 3 5 Promoter 5 3 3 55 RNA polymerase II Transcription factors RNA transcript Transcription initiation complex Eukaryotic promoters1 Several transcription factors 2 Additional transcription factors 3
  • 28. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Elongation of the RNA Strand • As RNA polymerase moves along the DNA – It continues to untwist the double helix, exposing about 10 to 20 DNA bases at a time for pairing with RNA nucleotides
  • 29. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Termination of Transcription • The mechanisms of termination – Are different in prokaryotes and eukaryotes
  • 30. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Concept 17.3: Eukaryotic cells modify RNA after transcription • Enzymes in the eukaryotic nucleus – Modify pre-mRNA in specific ways before the genetic messages are dispatched to the cytoplasm
  • 31. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Alteration of mRNA Ends • Each end of a pre-mRNA molecule is modified in a particular way – The 5′ end receives a modified nucleotide cap – The 3′ end gets a poly-A tail Figure 17.9 A modified guanine nucleotide added to the 5 end 50 to 250 adenine nucleotides added to the 3 end Protein-coding segment Polyadenylation signal Poly-A tail3 UTR Stop codonStart codon 5 Cap 5 UTR AAUAAA AAA…AAA TRANSCRIPTION RNA PROCESSING DNA Pre-mRNA mRNA TRANSLATION Ribosome Polypeptide G P P P 5 3
  • 32. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Split Genes and RNA Splicing • RNA splicing – Removes introns and joins exons Figure 17.10 TRANSCRIPTION RNA PROCESSING DNA Pre-mRNA mRNA TRANSLATION Ribosome Polypeptide 5 Cap Exon Intron 1 5 30 31 Exon Intron 104 105 146 Exon 3 Poly-A tail Poly-A tail Introns cut out and exons spliced together Coding segment 5 Cap 1 146 3 UTR3 UTR Pre-mRNA mRNA
  • 33. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Is carried out by spliceosomes in some cases Figure 17.11 RNA transcript (pre-mRNA) Exon 1 Intron Exon 2 Other proteins Protein snRNA snRNPs Spliceosome Spliceosome components Cut-out intron mRNA Exon 1 Exon 2 5 5 5 1 2 3
  • 34. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Ribozymes • Ribozymes – Are catalytic RNA molecules that function as enzymes and can splice RNA
  • 35. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The Functional and Evolutionary Importance of Introns • The presence of introns – Allows for alternative RNA splicing
  • 36. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Proteins often have a modular architecture – Consisting of discrete structural and functional regions called domains • In many cases – Different exons code for the different domains in a protein Figure 17.12 Gene DNA Exon 1 Intron Exon 2 Intron Exon 3 Transcription RNA processing Translation Domain 3 Domain 1 Domain 2 Polypeptide
  • 37. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Concept 17.4: Translation is the RNA-directed synthesis of a polypeptide: a closer look
  • 38. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Molecular Components of Translation • A cell translates an mRNA message into protein – With the help of transfer RNA (tRNA)
  • 39. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Translation: the basic concept Figure 17.13 TRANSCRIPTION TRANSLATION DNA mRNA Ribosome Polypeptide Polypeptide Amino acids tRNA with amino acid attachedRibosome tRNA Anticodon mRNA Trp Phe Gly A G C A A A C C G U G G U U U G G C Codons5 3
  • 40. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Molecules of tRNA are not all identical – Each carries a specific amino acid on one end – Each has an anticodon on the other end
  • 41. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The Structure and Function of Transfer RNA A C C • A tRNA molecule – Consists of a single RNA strand that is only about 80 nucleotides long – Is roughly L-shaped Figure 17.14a Two-dimensional structure. The four base-paired regions and three loops are characteristic of all tRNAs, as is the base sequence of the amino acid attachment site at the 3 end. The anticodon triplet is unique to each tRNA type. (The asterisks mark bases that have been chemically modified, a characteristic of tRNA.) (a) 3 C C A C G C U U A A GACAC CU * G C * * G U G U *CU * G AG G U * *A * A A G U C A G A C C * C G A G A G G G * * GA CUC*A U U U A G G C G 5 Amino acid attachment site Hydrogen bonds Anticodon A
  • 42. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Figure 17.14b (b) Three-dimensional structure Symbol used in this book Amino acid attachment site Hydrogen bonds Anticodon Anticodon A AG 5 3 3 5 (c)
  • 43. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • A specific enzyme called an aminoacyl-tRNA synthetase – Joins each amino acid to the correct tRNA Figure 17.15 Amino acid ATP Adenosine Pyrophosphate Adenosine Adenosine Phosphates tRNA P P P P P Pi Pi Pi P AMP Aminoacyl tRNA (an “activated amino acid”) Aminoacyl-tRNA synthetase (enzyme) Active site binds the amino acid and ATP. 1 ATP loses two P groups and joins amino acid as AMP. 2 3 Appropriate tRNA covalently Bonds to amino Acid, displacing AMP. Activated amino acid is released by the enzyme. 4
  • 44. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Ribosomes • Ribosomes – Facilitate the specific coupling of tRNA anticodons with mRNA codons during protein synthesis
  • 45. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • The ribosomal subunits – Are constructed of proteins and RNA molecules named ribosomal RNA or rRNA Figure 17.16a TRANSCRIPTION TRANSLATION DNA mRNA Ribosome Polypeptide Exit tunnel Growing polypeptide tRNA molecules E P A Large subunit Small subunit mRNA Computer model of functioning ribosome. This is a model of a bacterial ribosome, showing its overall shape. The eukaryotic ribosome is roughly similar. A ribosomal subunit is an aggregate of ribosomal RNA molecules and proteins. (a) 5 3
  • 46. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • The ribosome has three binding sites for tRNA – The P site – The A site – The E site Figure 17.16b E P A P site (Peptidyl-tRNA binding site) E site (Exit site) mRNA binding site A site (Aminoacyl- tRNA binding site) Large subunit Small subunit Schematic model showing binding sites. A ribosome has an mRNA binding site and three tRNA binding sites, known as the A, P, and E sites. This schematic ribosome will appear in later diagrams. (b)
  • 47. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Figure 17.16c Amino end Growing polypeptide Next amino acid to be added to polypeptide chain tRNA mRNA Codons 3 5 Schematic model with mRNA and tRNA. A tRNA fits into a binding site when its anticodon base-pairs with an mRNA codon. The P site holds the tRNA attached to the growing polypeptide. The A site holds the tRNA carrying the next amino acid to be added to the polypeptide chain. Discharged tRNA leaves via the E site. (c)
  • 48. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Building a Polypeptide • We can divide translation into three stages – Initiation – Elongation – Termination
  • 49. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Ribosome Association and Initiation of Translation • The initiation stage of translation – Brings together mRNA, tRNA bearing the first amino acid of the polypeptide, and two subunits of a ribosome Large ribosomal subunit The arrival of a large ribosomal subunit completes the initiation complex. Proteins called initiation factors (not shown) are required to bring all the translation components together. GTP provides the energy for the assembly. The initiator tRNA is in the P site; the A site is available to the tRNA bearing the next amino acid. 2 Initiator tRNA mRNA mRNA binding site Small ribosomal subunit Translation initiation complex P site GDPGTP Start codon A small ribosomal subunit binds to a molecule of mRNA. In a prokaryotic cell, the mRNA binding site on this subunit recognizes a specific nucleotide sequence on the mRNA just upstream of the start codon. An initiator tRNA, with the anticodon UAC, base-pairs with the start codon, AUG. This tRNA carries the amino acid methionine (Met). 1 Met Met U A C A U G E A 3 5 5 3 35 35 Figure 17.17
  • 50. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Elongation of the Polypeptide Chain • In the elongation stage of translation – Amino acids are added one by one to the preceding amino acid Figure 17.18 Amino end of polypeptide mRNA Ribosome ready for next aminoacyl tRNA E P A E P A E P A E P A GDP GTP GTP GDP 2 2 site site5 3 TRANSCRIPTION TRANSLATION DNA mRNA Ribosome Polypeptide Codon recognition. The anticodon of an incoming aminoacyl tRNA base-pairs with the complementary mRNA codon in the A site. Hydrolysis of GTP increases the accuracy and efficiency of this step. 1 Peptide bond formation. An rRNA molecule of the large subunit catalyzes the formation of a peptide bond between the new amino acid in the A site and the carboxyl end of the growing polypeptide in the P site. This step attaches the polypeptide to the tRNA in the A site. 2 Translocation. The ribosome translocates the tRNA in the A site to the P site. The empty tRNA in the P site is moved to the E site, where it is released. The mRNA moves along with its bound tRNAs, bringing the next codon to be translated into the A site. 3
  • 51. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Termination of Translation • The final stage of translation is termination – When the ribosome reaches a stop codon in the mRNA Figure 17.19 Release factor Free polypeptide Stop codon (UAG, UAA, or UGA) 5 3 3 5 3 5 When a ribosome reaches a stop codon on mRNA, the A site of the ribosome accepts a protein called a release factor instead of tRNA. 1 The release factor hydrolyzes the bond between the tRNA in the P site and the last amino acid of the polypeptide chain. The polypeptide is thus freed from the ribosome. 2 3 The two ribosomal subunits and the other components of the assembly dissociate.
  • 52. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Polyribosomes • A number of ribosomes can translate a single mRNA molecule simultaneously – Forming a polyribosome Figure 17.20a, b Growing polypeptides Completed polypeptide Incoming ribosomal subunits Start of mRNA (5 end) End of mRNA (3 end) Polyribosome An mRNA molecule is generally translated simultaneously by several ribosomes in clusters called polyribosomes. (a) Ribosomes mRNA This micrograph shows a large polyribosome in a prokaryotic cell (TEM). 0.1 µm (b)
  • 53. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Completing and Targeting the Functional Protein • Polypeptide chains – Undergo modifications after the translation process
  • 54. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Protein Folding and Post-Translational Modifications • After translation – Proteins may be modified in ways that affect their three-dimensional shape
  • 55. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Targeting Polypeptides to Specific Locations • Two populations of ribosomes are evident in cells – Free and bound • Free ribosomes in the cytosol – Initiate the synthesis of all proteins
  • 56. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Proteins destined for the endomembrane system or for secretion – Must be transported into the ER – Have signal peptides to which a signal- recognition particle (SRP) binds, enabling the translation ribosome to bind to the ER
  • 57. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Figure 17.21 Ribosome mRNA Signal peptide Signal- recognition particle (SRP) SRP receptor protein Translocation complex CYTOSOL Signal peptide removed ER membrane Protein ERLUMEN • The signal mechanism for targeting proteins to the ER Polypeptide synthesis begins on a free ribosome in the cytosol. 1 An SRP binds to the signal peptide, halting synthesis momentarily. 2 The SRP binds to a receptor protein in the ER membrane. This receptor is part of a protein complex (a translocation complex) that has a membrane pore and a signal-cleaving enzyme. 3 The SRP leaves, and the polypeptide resumes growing, meanwhile translocating across the membrane. (The signal peptide stays attached to the membrane.) 4 The signal- cleaving enzyme cuts off the signal peptide. 5 The rest of the completed polypeptide leaves the ribosome and folds into its final conformation. 6
  • 58. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Concept 17.5: RNA plays multiple roles in the cell: a review • RNA – Can hydrogen-bond to other nucleic acid molecules – Can assume a specific three-dimensional shape – Has functional groups that allow it to act as a catalyst
  • 59. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Types of RNA in a Eukaryotic Cell Table 17.1
  • 60. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Concept 17.6: Comparing gene expression in prokaryotes and eukaryotes reveals key differences • Prokaryotic cells lack a nuclear envelope – Allowing translation to begin while transcription is still in progress Figure 17.22 DNA Polyribosome mRNA Direction of transcription 0.25 mRNA polymerase Polyribosome Ribosome DNA mRNA (5 end) RNA polymerase Polypeptide (amino end)
  • 61. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • In a eukaryotic cell – The nuclear envelope separates transcription from translation – Extensive RNA processing occurs in the nucleus
  • 62. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Concept 17.7: Point mutations can affect protein structure and function • Mutations – Are changes in the genetic material of a cell • Point mutations – Are changes in just one base pair of a gene
  • 63. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • The change of a single nucleotide in the DNA’s template strand – Leads to the production of an abnormal protein Figure 17.23 In the DNA, the mutant template strand has an A where the wild-type template has a T. The mutant mRNA has a U instead of an A in one codon. The mutant (sickle-cell) hemoglobin has a valine (Val) instead of a glutamic acid (Glu). Mutant hemoglobin DNAWild-type hemoglobin DNA mRNA mRNA Normal hemoglobin Sickle-cell hemoglobin Glu Val C T T C A T G A A G U A 3 5 3 5 5 35 3
  • 64. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Types of Point Mutations • Point mutations within a gene can be divided into two general categories – Base-pair substitutions – Base-pair insertions or deletions
  • 65. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Substitutions • A base-pair substitution – Is the replacement of one nucleotide and its partner with another pair of nucleotides – Can cause missense or nonsense Figure 17.24 Wild type A U G A A G U U U G G C U A A mRNA 5 Protein Met Lys Phe Gly Stop Carboxyl end Amino end 3 A U G A A G U U U G G U U A A Met Lys Phe Gly Base-pair substitution No effect on amino acid sequence U instead of C Stop A U G A A G U U U A G U U A A Met Lys Phe Ser Stop A U G U A G U U U G G C U A A Met Stop Missense A instead of G Nonsense U instead of A
  • 66. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Insertions and Deletions • Insertions and deletions – Are additions or losses of nucleotide pairs in a gene – May produce frameshift mutations Figure 17.25 mRNA Protein Wild type A U G A A G U U U G G C U A A 5 Met Lys Phe Gly Amino end Carboxyl end Stop Base-pair insertion or deletion Frameshift causing immediate nonsense A U G U A A G U U U G G C U A A U G A A G U U G G C U A A A U G U U U G G C U A A Met Stop U Met Lys Leu Ala Met Phe Gly Stop MissingA A G Missing Extra U Frameshift causing extensive missense Insertion or deletion of 3 nucleotides: no frameshift but extra or missing amino acid 3
  • 67. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Mutagens • Spontaneous mutations – Can occur during DNA replication, recombination, or repair
  • 68. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • Mutagens – Are physical or chemical agents that can cause mutations
  • 69. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings What is a gene? revisiting the question • A gene – Is a region of DNA whose final product is either a polypeptide or an RNA molecule
  • 70. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings • A summary of transcription and translation in a eukaryotic cell Figure 17.26 TRANSCRIPTION RNA is transcribed from a DNA template. DNA RNA polymerase RNA transcript RNA PROCESSING In eukaryotes, the RNA transcript (pre- mRNA) is spliced and modified to produce mRNA, which moves from the nucleus to the cytoplasm. Exon Poly-A RNA transcript (pre-mRNA) Intron NUCLEUS Cap FORMATION OF INITIATION COMPLEX After leaving the nucleus, mRNA attaches to the ribosome. CYTOPLASM mRNA Poly-A Growing polypeptide Ribosomal subunits Cap Aminoacyl-tRNA synthetase Amino acid tRNA AMINO ACID ACTIVATION Each amino acid attaches to its proper tRNA with the help of a specific enzyme and ATP. Activated amino acid TRANSLATION A succession of tRNAs add their amino acids to the polypeptide chain as the mRNA is moved through the ribosome one codon at a time. (When completed, the polypeptide is released from the ribosome.) Anticodon A CC A A A UG GUU UA U G UACE A Ribosome 1 Poly-A 5 5 3 Codon 2 3 4 5