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Chapter 17
From Gene to Protein
Transcription and Translation
Overview: The Flow of Genetic
Information
• The information content of DNA is in the form
of specific sequences of nucleotides
• The DNA inherited by an organism leads to
specific traits by dictating the synthesis of
proteins
• Gene expression, the process by which DNA
directs protein synthesis, includes two stages:
transcription and translation
• The ribosome is part of the cellular machinery
for translation, polypeptide synthesis
Evidence from the Study of Metabolic
Defects
• In 1902, British physician Archibald Garrod first
suggested that genes dictate phenotypes through
enzymes that catalyze specific chemical reactions
• He thought symptoms of an inherited disease
reflect an inability to synthesize a certain enzyme
• Linking genes to enzymes required understanding
that cells synthesize and degrade molecules in a
series of steps, a metabolic pathway
Nutritional Mutants in Neurospora:
Scientific Inquiry
• Beadle and Tatum exposed bread mold to X-rays,
creating mutants that were unable to survive on
minimal medium as a result of inability to
synthesize certain molecules
• Using crosses, they identified three classes of
arginine-deficient mutants, each lacking a different
enzyme necessary for synthesizing arginine
• They developed a “one gene–one enzyme”
hypothesis, which states that each gene dictates
production of a specific enzyme
Figure 17.2
Precursor
Enzyme A
Enzyme B
Enzyme C
Ornithine
Citrulline
Arginine
No growth:
Mutant cells
cannot grow
and divide
Growth:
Wild-type
cells growing
and dividing
Control: Minimal medium
Results Table
Wild type
Minimal
medium
(MM)
(control)
MM +
ornithine
MM +
citrulline
MM +
arginine
(control)
Summary
of results
Can grow with
or without any
supplements
Gene
(codes for
enzyme) Wild type
Precursor
Ornithine
Gene A
Gene B
Gene C
Enzyme A
Enzyme B
Enzyme C
Enzyme A
Enzyme B
Enzyme C
Citrulline
Arginine
Precursor
Ornithine
Citrulline
Arginine
Precursor
Ornithine
Citrulline
Arginine
Precursor
Ornithine
Citrulline
Arginine
Enzyme A
Enzyme B
Enzyme C
Enzyme A
Enzyme B
Enzyme C
Class I mutants
(mutation in
gene A)
Class II mutants
(mutation in
gene B)
Class III mutants
(mutation in
gene C)
Can grow on
ornithine,
citrulline,
or arginine
Can grow only
on citrulline or
arginine
Require arginine
to grow
Class I mutants Class II mutants Class III mutants
Classes of Neurospora crassa
Condition
The Products of Gene Expression: A
Developing Story
• Some proteins aren’t enzymes, so researchers later
revised the hypothesis: one gene–one protein
• Many proteins are composed of several
polypeptides, each of which has its own gene
• Therefore, Beadle and Tatum’s hypothesis is now
restated as one gene–one polypeptide
Basic Principles of Transcription and
Translation
• Transcription is the synthesis of RNA under the
direction of DNA
• Transcription produces messenger RNA (mRNA)
• Translation is the synthesis of a polypeptide,
which occurs under the direction of mRNA
• Ribosomes are the sites of translation
• In prokaryotes, mRNA produced by transcription
is immediately translated without more
processing--coupled
• In a eukaryotic cell, the nuclear envelope
separates transcription from translation
• Eukaryotic RNA transcripts are modified through
RNA processing to yield finished mRNA
• Cells are governed by a cellular chain of
command: DNA → RNA → protein
Figure 17.3
Nuclear
envelope
CYTOPLASM
DNA
Pre-mRNA
mRNA
RibosomeTRANSLATION
(b) Eukaryotic cell
NUCLEUS
RNA PROCESSING
TRANSCRIPTION
(a) Bacterial cell
Polypeptide
DNA
mRNA
Ribosome
CYTOPLASM
TRANSCRIPTION
TRANSLATION
Polypeptide
The Genetic Code
• How are the instructions for assembling amino
acids into proteins encoded into DNA?
• There are 20 amino acids, but there are only four
nucleotide bases in DNA
• So how many bases correspond to an amino acid?
Codons: Triplets of Bases
• The flow of information from gene to protein is
based on a triplet code: a series of
nonoverlapping, three-nucleotide words
• These triplets are the smallest units of uniform
length that can code for all the amino acids
• Example: AGT at a particular position on a DNA
strand results in the placement of the amino
acid serine at the corresponding position of the
polypeptide to be produced
• During transcription, a DNA strand called the
template strand provides a template for
ordering the sequence of nucleotides in an RNA
transcript
• During translation, the mRNA base triplets,
called codons, are read in the 5′ to 3′ direction
• Each codon specifies the amino acid to be
placed at the corresponding position along a
polypeptide
Figure 17.4
A C C A A A C C G A G T
ACTTTT CGGGGT
U G G U U U G G C CU A
SerGlyPheTrp
Codon
TRANSLATION
TRANSCRIPTION
Protein
mRNA 5′
5′
3′
Amino acid
DNA
template
strand
5′
3′
3′
Cracking the Code
• All 64 codons were deciphered by the mid-1960s
• The genetic code is redundant but not ambiguous;
no codon specifies more than one amino acid
• Codons must be read in the correct reading frame
(correct groupings) in order for the specified
polypeptide to be produced
Figure 17.5
Second mRNA base
ThirdmRNAbase(3′endofcodon)
FirstmRNAbase(5′endofcodon)
UUU
UUC
UUA
UUG
Phe
Leu
Leu
Ile
Val
CUU
CUC
CUA
CUG
AUU
AUC
AUA
AUG
GUU
GUC
GUA
GUG
UCU
UCC
UCA
UCG
CCU
CCC
CCA
CCG
ACU
ACC
ACA
ACG
GCU
GCC
GCA
GCG GAG
GAA
GAC
GAU
AAG
AAA
AAC
AAU
CAG
CAA
CAC
CAU
Ser
Pro
Thr
Ala
Glu
Asp
Lys
Asn
Gln
His
Tyr Cys
Trp
Arg
Ser
Arg
Gly
GGG
GGA
GGC
GGU
AGG
AGA
AGC
AGU
CGG
CGA
CGC
CGU
UGG
UGA
UGC
UGUUAU
UAC
UAA
UAG Stop
Stop Stop
Met or
start
U
C
A
G
U
C
A
G
U
C
A
G
U
C
A
G
U C A G
G
A
C
U
Evolution of the Genetic Code
• The genetic code is nearly universal, shared by
the simplest bacteria to the most complex
animals
• Genes can be transcribed and translated after
being transplanted from one species to another
Pig expressing a
jellyfish
gene
(b)Tobacco plant
expressing
a firefly gene
(a)
Molecular Components of
Transcription
• RNA synthesis is catalyzed by RNA polymerase,
which pries the DNA strands apart and hooks
together the RNA nucleotides
• RNA synthesis follows the same base-pairing rules
as DNA, except uracil substitutes for thymine
• The DNA sequence where RNA polymerase
attaches is called the promoter; in prokaryotes,
the sequence signaling the end of transcription
is called the terminator
• The stretch of DNA that is transcribed is called a
transcription unit
Synthesis of an RNA Transcript
• The three stages of transcription:
– Initiation
– Elongation
– Termination
Figure 17.7-3 Promoter Transcription unit
RNA polymerase
Start point
1
Template strand of DNARNA
transcript
Unwound
DNA
Rewound
DNA
RNA
transcript
Direction of
transcription
(“downstream”)
Completed RNA transcript
Initiation
Elongation
Termination
2
3
5′
3′
5′
5′
3′
5′
3′
5′
3′
5′
5′
3′
3′
5′
3′
3′
5′
3′
5′
3′
Figure 17.9 Nontemplate
strand of DNA
5′
3′
3′ end
5′
3′
5′
Direction of transcription
RNA
polymerase
Template
strand of DNA
Newly made
RNA
RNA nucleotides
A
A
A
A
A AA
A
A
C
C
G G T T
T
C
C CU
U
C CT
T
C
A
T
T
G
G
U
RNA Polymerase Binding and Initiation
of Transcription
• Promoters signal the initiation of RNA synthesis
• Transcription factors mediate the binding of RNA
polymerase and initiation of transcription in
eukaryotes
• The completed assembly of transcription factors
and RNA polymerase II bound to a promoter is
called a transcription initiation complex
• A promoter called a TATA box is crucial in forming
the initiation complex in eukaryotes
Promoter
Nontemplate strand
15′
3′
5′
3′
Start point
RNA polymerase II
Template
strand
TATA box
Transcription
factors
DNA
3′
5′
3′
5′
3′
5′
2
3
Transcription factors
RNA transcript
Transcription initiation complex
3′5′
5′
3′
A eukaryotic
promoter
Several
transcription
factors bind
to DNA.
Transcription
initiation
complex
forms.
T A T AAAA
TA AT T T T
Elongation of the RNA Strand
• As RNA polymerase moves along the DNA, it
untwists the double helix, 10 to 20 bases at a time
• Transcription progresses at a rate of 40 nucleotides
per second in eukaryotes
• A gene can be transcribed simultaneously by
several RNA polymerases
Termination of Transcription
• The mechanisms of termination are different in
prokaryotes and eukaryotes
• In prokaryotes, the polymerase stops
transcription at the end of the terminator
• In eukaryotes, RNA polymerase II transcribes
the polyadenylation signal sequence; the RNA
transcript is released 10–35 nucleotides past
this polyadenylation sequence
Concept 17.3: Eukaryotic cells modify
RNA after transcription
• Enzymes in the eukaryotic nucleus modify pre-
mRNA (RNA processing) before the messages
are dispatched to the cytoplasm
• During RNA processing, both ends of the
primary transcript are usually altered (5’ cap
and 3’ polyA tail addition)
• Also, usually some interior parts of the molecule
are cut out, and the other parts spliced together
(RNA splicing)
Figure 17.10
A modified guanine
nucleotide added to
the 5′ end
Region that includes
protein-coding segments
5′
5′
Cap
5′
UTR
Start
codon
Stop
codon
G P P P
3′
UTR
3′
AAUAAA AAA AAA
Poly-A tail
Polyadenylation
signal
50–250 adenine
nucleotides added
to the 3′ end
…
These modifications share several functions:
• They seem to facilitate the export of mRNA
• They protect mRNA from hydrolytic
enzymes
Split Genes and RNA Splicing
• Most eukaryotic genes and their RNA transcripts
have long noncoding stretches of nucleotides
that lie between coding regions
• These noncoding regions are called intervening
sequences, or introns
• The other regions are called exons because they
are eventually expressed, usually translated into
amino acid sequences
• RNA splicing removes introns and joins exons,
creating an mRNA molecule with a continuous
coding sequence
Figure 17.11
Pre-mRNA Intron Intron
Introns cut out
and exons
spliced together
Poly-A tail5′ Cap
5′ Cap Poly-A tail
1–30 31–104 105–146
1–146
3′ UTR5′ UTR
Coding
segment
mRNA
• In some cases, RNA splicing is carried out by spliceosomes
• Spliceosomes consist of a variety of proteins and several
small nuclear ribonucleoproteins (snRNPs) that recognize
the splice sites
Figure 17.12
Spliceosome Small RNAs
Exon 2
Cut-out
intron
Spliceosome
components
mRNA
Exon 1 Exon 2
Pre-mRNA
Exon 1
Intron
5′
5′
Ribozymes
• Ribozymes are catalytic RNA molecules
• Here they function as enzymes that can splice
RNA
• The discovery of ribozymes rendered obsolete
the belief that all biological catalysts were
proteins
• Three properties of RNA enable it to function
as an enzyme
– It can form a three-dimensional structure because
of its ability to base-pair with itself
– Some bases in RNA contain functional groups
that may participate in catalysis
– RNA may hydrogen-bond with other nucleic
acid molecules
The Functional and Evolutionary
Importance of Introns
• Some introns contain sequences that may
regulate gene expression
• Some genes can encode more than one kind
of polypeptide, depending on which segments
are treated as exons during splicing
• This is called alternative RNA splicing
• Consequently, the number of different
proteins
an organism can produce is much greater than
its number of genes
• Proteins often have a modular architecture
consisting of discrete regions called domains
• In many cases, different exons code for the
different domains in a protein
Figure 17.13
Gene
DNA
Transcription
RNA processing
Translation
Domain 3
Exon 1 Exon 3Exon 2 IntronIntron
Domain 2
Domain 1
Polypeptide
Molecular Components of Translation
• A cell translates an mRNA message into protein
with the help of transfer RNA (tRNA) &
ribosomes
• tRNAs transfer amino acids to the growing
polypeptide in a ribosome
• Molecules of tRNA are not identical:
– Each carries a specific amino acid on one end
– Each has an anticodon on the other end; the
anticodon base-pairs with a complementary codon
on mRNA
Figure 17.14
Polypeptide
Amino
acids
tRNA with
amino acid
attached
Ribosome
tRNA
Anticodon
Codons
mRNA
5′
U U U UG G G G C
A
C C
A A A
C
C
G
Phe
Trp
3′
Gly
The Structure and Function of Transfer
RNA
• A tRNA molecule consists of a single RNA
strand that is only about 80 nucleotides long
• Flattened into one plane to reveal its base
pairing, a tRNA molecule looks like a cloverleaf
• Amino acid at one end of sock and anticodon at
the other
• Because of hydrogen bonds, tRNA actually
twists and folds into a three-dimensional
molecule
• tRNA is roughly L-shaped
A
C
C
Figure 17.15
Amino acid
attachment
site
Amino acid
attachment site
5′
3′
A
C
C
A
C
G
C
U
U
A
G
G
C
G
A
U
U
U
A
A
GAA CC
CU
*
**
C
G
G U UG
C
*
*
*
*C CU
A G
G
G
G
A
GAGC
C
C
*U
* G A
G
G
U
*
*
*
A
A
A
G
C
U
G
A
A
Hydrogen
bonds
Hydrogen
bonds
Anticodon Anticodon
Symbol used
in this book
Three-dimensional
structure
(a) Two-dimensional structure (b) (c)
Anticodon
5′3′
A A G
3′
5′
• Accurate translation requires two steps:
– First step: a correct match between a tRNA and an
amino acid, done by the enzyme aminoacyl-tRNA
synthetase
– Second step: a correct match between the tRNA
anticodon and an mRNA codon
Flexible pairing at the third base of a codon is called
wobble and allows some tRNAs to bind to more
than one codon
Figure 17.16-3
Aminoacyl-tRNA
synthetase
1 Amino acid
and tRNA
enter active
site.
Tyr-tRNA
Tyrosine (Tyr)
(amino acid)
Tyrosyl-tRNA
synthetase
Complementary
tRNA anticodon
A AU
tRNA
Amino
acid
Using ATP,
synthetase
catalyzes
covalent
bonding.
23
ATP
AMP + 2 P i
Aminoacyl
tRNA
released.
Computer model
Ribosomes
• Ribosomes facilitate specific coupling of tRNA
anticodons with mRNA codons in protein synthesis
• The two ribosomal subunits (large and small) are
made of proteins and ribosomal RNA (rRNA)
Figure
17.17
Exit tunnel
Large
subunit
Small
subunit
mRNA 3′
5′
E P A
tRNA
molecules
Growing
polypeptide
(a) Computer model of functioning ribosome
Growing
polypeptide Next amino
acid to be
added to
polypeptide
chain
tRNA
3′
5′
mRNA
Amino end
Codons
E
(c) Schematic model with mRNA and tRNA(b) Schematic model showing binding sites
Small
subunit
Large
subunit
Exit tunnel
A site (Aminoacyl-
tRNA binding site)
P site (Peptidyl-tRNA
binding site)
E site
(Exit site)
mRNA
binding
site
E P A
• A ribosome has three binding sites for tRNA:
– The P site holds the tRNA that carries the growing
polypeptide chain
– The A site holds the tRNA that carries the next
amino acid to be added to the chain
– The E site is the exit site, where uncharged tRNAs
leave the ribosome
Building a Polypeptide
• The three stages of translation:
–Initiation
–Elongation
–Termination
–All 3 stages require protein factors and some
require energy in the form of GTP.
Ribosome Association and
Initiation of Translation
• Initiation brings together mRNA, a tRNA with
the first amino acid, and the two ribosomal
subunits
• First, a small ribosomal subunit binds with
mRNA and a special initiator tRNA
• Then the small subunit moves along the
mRNA until it reaches the start codon (AUG)
• Proteins called initiation factors bring in the
large subunit that completes the translation
initiation complex
Figure 17.18
1 2
P site
3′
Large
ribosomal
subunit
Translation initiation complex
Large ribosomal subunit
completes the initiation
complex.
Small ribosomal subunit binds
to mRNA.
mRNA binding site
Small
ribosomal
subunit
Initiator
tRNA
Start codon
3′5′
E A
3′
5′
GTP GDP
P i
+
3′ 5′
5′
U A C
GA UMet
Met
mRNA
Elongation of the Polypeptide
Chain
• During elongation, amino acids are added one
by one to the C-terminus of the growing chain
• Each addition involves proteins called
elongation factors and occurs in three steps:
codon recognition, peptide bond formation,
and translocation
• Energy expenditure occurs in the first (docking
in A site) and third steps(translocation)
• Translation proceeds along the mRNA in a
5 → 3 direction′ ′
Amino end
of polypeptide
Codon
recognition
1
3′
5′
E
P A
sitesite
E
P A
mRNA
GTP
P iGDP +
Amino end
of polypeptide
Codon
recognition
1
3′
5′
E
P A
sitesite
E
P A
mRNA
GTP
P i
2
GDP +
E
P A
Peptide bond
formation
Amino end
of polypeptide
Codon
recognition
1
3′
5′
E
P A
sitesite
E
P A
mRNA
GTP
P i
23 GTP
P i
GDP +
GDP +
Translocation
E
P A
Peptide bond
formation
E
P A
Ribosome ready for
next aminoacyl tRNA
Termination of Translation
• Termination occurs when a stop codon in the
mRNA reaches the A site of the ribosome
• The A site accepts a protein called a release
factor
• The release factor causes the addition of a
water molecule instead of an amino acid
• This reaction releases the polypeptide, and
the translation assembly comes apart
Figure 17.20-3
31 2
Release
factor
3′
5′5′
3′
Stop codon
(UAG, UAA, or UGA)
Ribosome reaches a
stop codon on mRNA.
Release factor
promotes hydrolysis.
Ribosomal subunits
and other components
dissociate.
Free
polypeptide
3′
5′
2 GTP
2 GDP + 2 P i
Polyribosomes
• A number of ribosomes can translate a single
mRNA simultaneously, forming a polyribosome
• Polyribosomes enable a cell to make many
copies of a polypeptide very quickly
LE 17-20
Ribosomes
mRNA
0.1 µm
This micrograph shows a large polyribosome in a prokaryotic cell (TEM).
An mRNA molecule is generally translated simultaneously
by several ribosomes in clusters called polyribosomes.
Incoming
ribosomal
subunits
Growing
polypeptides
End of
mRNA
(3′ end)
Start of
mRNA
(5′ end)
Polyribosome
Completed
polypeptides
Completing and Targeting the
Functional Protein
• Often translation is not sufficient to make a
functional protein
• Polypeptide chains are modified after
translation
• Completed proteins are targeted to specific
sites in the cell
Targeting Polypeptides to Specific
Locations
• Two populations of ribosomes are evident in
cells: free ribsomes (in the cytosol) and bound
ribosomes (attached to the ER)
• Free ribosomes mostly synthesize proteins that
function in the cytosol
• Bound ribosomes make proteins of the
endomembrane system and proteins that are
secreted from the cell (rough ER)
• Ribosomes are identical and can switch from
free to bound
• Polypeptide synthesis always begins in the
cytosol
• Synthesis finishes in the cytosol unless the
polypeptide signals the ribosome to attach to
the ER
• Polypeptides destined for the ER or for
secretion are marked by a signal peptide
• A signal-recognition particle (SRP) binds to the
signal peptide
• The SRP brings the signal peptide and its
ribosome to the ER
Figure 17.21
Protein
ER
membrane
Signal
peptide
removed
SRP receptor
protein
Translocation complex
ER LUMEN
CYTOSOL
SRP
Signal
peptide
mRNA
Ribosome
Polypeptide
synthesis
begins.
SRP
binds to
signal
peptide.
SRP
binds to
receptor
protein.
SRP
detaches
and
polypeptide
synthesis
resumes.
Signal-
cleaving
enzyme cuts
off signal
peptide.
Completed
polypeptide
folds into
final
conformation.
1 2 3 4 5 6
• A bacterial cell ensures a streamlined process
by coupling transcription and translation
• In this case the newly made protein can
quickly diffuse to its site of function
RNA polymerase
Polyribosome
DNA
mRNA
0.25 µm
Ribosome
DNA
mRNA (5′ end)
Polyribosome
Polypeptide
(amino end)
Direction of
transcriptionRNA
polymerase
• In eukaryotes, the nuclear envelop separates
the processes of transcription and translation
• RNA undergoes processes before leaving
the nucleus
DNA
RNA
polymerase
RNA transcript
(pre-mRNA)
Intron
Exon
Aminoacyl-tRNA
synthetase
Amino
acid
tRNA
Aminoacyl
(charged)
tRNA
mRNA
CYTOPLASM
NUCLEUS
RNA
transcript
3′
5′
Poly-A
5′ Cap
5′ C
ap
TRANSCRIPTION
RNA
PROCESSING
Poly-A
Poly-A
AMINO ACID
ACTIVATION
TRANSLATION
Ribosomal
subunits
E
P
A
E A
A
C C
U U U U U GGG
A A A
Anticodon
Codon
Ribosome
C
A
U
3′
A
Concept 17.5: RNA plays multiple
roles in the cell: a review
Type of RNA Functions
Messenger
RNA (mRNA)
Carries information specifying
amino acid sequences of
proteins from DNA to ribosomes
Transfer RNA
(tRNA)
Serves as adapter molecule in
protein synthesis; translates
mRNA codons into amino acids
Ribosomal
RNA (rRNA)
Plays catalytic (ribozyme) roles
and structural roles in ribosomes
Type of RNA Functions
Primary
transcript
Serves as a precursor to mRNA,
rRNA, or tRNA, before being
processed by splicing or
cleavage
Small nuclear
RNA (snRNA)
Plays structural and catalytic
roles in spliceosomes
SRP RNA Is a component of the the signal-
recognition particle (SRP)
Video: protein synthesis
Concept 17.7: Point mutations can
affect protein structure and function
• Mutations are changes in the genetic material
of a cell or virus
• Point mutations are chemical changes in just
one base pair of a gene
• The change of a single nucleotide in a DNA
template strand leads to production of an
abnormal protein
Figure 17.25
Wild-type β-globin Sickle-cell β-globin
Mutant β-globin DNAWild-type β-globin DNA
mRNA mRNA
Normal hemoglobin Sickle-cell hemoglobin
Val
U GG
A CC
G GT
Glu
G GA
G GA
C CT3′
5′
5′ 5′
5′
5′
3′
3′
3′ 3′
3′
5′
Types of Point Mutations
• Point mutations within a gene can be divided
into two general categories
– Base-pair substitutions
– Base-pair insertions or deletions
Substitutions
• A base-pair substitution replaces one nucleotide
and its partner with another pair of nucleotides
• Base-pair substitution can cause missense or
nonsense mutations
• Missense mutations still code for an amino acid,
but not necessarily the right amino acid
• Nonsense mutations change an amino acid
codon into a stop codon, nearly always leading
to a nonfunctional protein
• Missense mutations are more common
Wild type
Stop
5′
3′
3′
Stop
3′
5′
5′
Met
(b) Nucleotide-pair insertion or deletion
Carboxyl endAmino end
mRNA
Protein
3′
5′
5′
DNA template strand
(a) Nucleotide-pair substitution
A instead of G
U instead of C
5′
3′
3′
Stop
Extra A
Extra U
Frameshift (1 nucleotide-pair insertion)
missing
missing
Frameshift (1 nucleotide-pair deletion)
missing
3′
5′
5′
missing
3′
5′
3′
Met Lys Leu Ala
3′
5′
3′
T A C T T C A A A C C G A T TA
TA G T A A A AC TTTTG G G
G G C U A AAAA U U UUUGG
A U G A A G G G C UUU
U
A A
AATCGGTTT GG A AA
T A C T T C A A C C G A T T
A
Met Lys Phe Gly
T A C C C C GA A A A
A AAAA
A U G
G G G G
T T T T
TTTTT C
G G G C UUUUAA A A
3′
5′
5′
T
A A A A A
T T T TA A A A AACCCC
T T T T TTGGGG
G G G GUUU U A AA A AU
Met Lys Phe Gly Stop
U
Silent
3′
5′
5′
5′
3′
3′
Stop
Stop Stop
3 nucleotide-pair deletionNonsense
A instead of T
U instead of A
5′
5′
5′
3′
3′
3′
Met Lys Phe Ser
Met Met Phe Gly
A
T
A
A
T
A
A
T
A
T instead of C
A instead of G
Missense
U G G G G A AU U U U UU A
A A A
T T TA A A A AGCCCC A
T T T T T TCGGGG
3′
5′
5′
5′
3′
3′
T T
A A
AA
A A G
U U U U UCGGG
T T T T T
AGCC
CGGG
C
CTT
AAAA
T T T TT
A
AA AA C C C GA
A A A ATTTT GGGT C
A A AAUCGGGU U U U A
Mutagens
• Spontaneous mutations can occur during DNA
replication, recombination, or repair
• Mutagens are physical or chemical agents that
can cause mutations
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
LE 17-26
TRANSCRIPTION
RNA PROCESSING
RNA
transcript
5′
Exon
NUCLEUS
FORMATION OF
INITIATION COMPLEX
CYTOPLASM
3′
DNA
RNA
polymerase
RNA transcript
(pre-mRNA)
Intron
Aminoacyl-tRNA
synthetase
Amino
acid
tRNA
AMINO ACID ACTIVATION
3′
mRNA
A
P
E Ribosomal
subunits
5′
Growing
polypeptide
E A
Activated
amino acid
Anticodon
TRANSLATION
Codon
Ribosome
Review compartments for eukaryotes

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Gene to Protein Flow

  • 1. Chapter 17 From Gene to Protein Transcription and Translation
  • 2. Overview: The Flow of Genetic Information • The information content of DNA is in the form of specific sequences of nucleotides • The DNA inherited by an organism leads to specific traits by dictating the synthesis of proteins • Gene expression, the process by which DNA directs protein synthesis, includes two stages: transcription and translation • The ribosome is part of the cellular machinery for translation, polypeptide synthesis
  • 3. Evidence from the Study of Metabolic Defects • In 1902, British physician Archibald Garrod first suggested that genes dictate phenotypes through enzymes that catalyze specific chemical reactions • He thought symptoms of an inherited disease reflect an inability to synthesize a certain enzyme • Linking genes to enzymes required understanding that cells synthesize and degrade molecules in a series of steps, a metabolic pathway
  • 4. Nutritional Mutants in Neurospora: Scientific Inquiry • Beadle and Tatum exposed bread mold to X-rays, creating mutants that were unable to survive on minimal medium as a result of inability to synthesize certain molecules • Using crosses, they identified three classes of arginine-deficient mutants, each lacking a different enzyme necessary for synthesizing arginine • They developed a “one gene–one enzyme” hypothesis, which states that each gene dictates production of a specific enzyme
  • 5. Figure 17.2 Precursor Enzyme A Enzyme B Enzyme C Ornithine Citrulline Arginine No growth: Mutant cells cannot grow and divide Growth: Wild-type cells growing and dividing Control: Minimal medium Results Table Wild type Minimal medium (MM) (control) MM + ornithine MM + citrulline MM + arginine (control) Summary of results Can grow with or without any supplements Gene (codes for enzyme) Wild type Precursor Ornithine Gene A Gene B Gene C Enzyme A Enzyme B Enzyme C Enzyme A Enzyme B Enzyme C Citrulline Arginine Precursor Ornithine Citrulline Arginine Precursor Ornithine Citrulline Arginine Precursor Ornithine Citrulline Arginine Enzyme A Enzyme B Enzyme C Enzyme A Enzyme B Enzyme C Class I mutants (mutation in gene A) Class II mutants (mutation in gene B) Class III mutants (mutation in gene C) Can grow on ornithine, citrulline, or arginine Can grow only on citrulline or arginine Require arginine to grow Class I mutants Class II mutants Class III mutants Classes of Neurospora crassa Condition
  • 6. The Products of Gene Expression: A Developing Story • Some proteins aren’t enzymes, so researchers later revised the hypothesis: one gene–one protein • Many proteins are composed of several polypeptides, each of which has its own gene • Therefore, Beadle and Tatum’s hypothesis is now restated as one gene–one polypeptide
  • 7. Basic Principles of Transcription and Translation • Transcription is the synthesis of RNA under the direction of DNA • Transcription produces messenger RNA (mRNA) • Translation is the synthesis of a polypeptide, which occurs under the direction of mRNA • Ribosomes are the sites of translation
  • 8. • In prokaryotes, mRNA produced by transcription is immediately translated without more processing--coupled • In a eukaryotic cell, the nuclear envelope separates transcription from translation • Eukaryotic RNA transcripts are modified through RNA processing to yield finished mRNA • Cells are governed by a cellular chain of command: DNA → RNA → protein
  • 9. Figure 17.3 Nuclear envelope CYTOPLASM DNA Pre-mRNA mRNA RibosomeTRANSLATION (b) Eukaryotic cell NUCLEUS RNA PROCESSING TRANSCRIPTION (a) Bacterial cell Polypeptide DNA mRNA Ribosome CYTOPLASM TRANSCRIPTION TRANSLATION Polypeptide
  • 10. The Genetic Code • How are the instructions for assembling amino acids into proteins encoded into DNA? • There are 20 amino acids, but there are only four nucleotide bases in DNA • So how many bases correspond to an amino acid?
  • 11. Codons: Triplets of Bases • The flow of information from gene to protein is based on a triplet code: a series of nonoverlapping, three-nucleotide words • These triplets are the smallest units of uniform length that can code for all the amino acids • Example: AGT at a particular position on a DNA strand results in the placement of the amino acid serine at the corresponding position of the polypeptide to be produced
  • 12. • During transcription, a DNA strand called the template strand provides a template for ordering the sequence of nucleotides in an RNA transcript • During translation, the mRNA base triplets, called codons, are read in the 5′ to 3′ direction • Each codon specifies the amino acid to be placed at the corresponding position along a polypeptide
  • 13. Figure 17.4 A C C A A A C C G A G T ACTTTT CGGGGT U G G U U U G G C CU A SerGlyPheTrp Codon TRANSLATION TRANSCRIPTION Protein mRNA 5′ 5′ 3′ Amino acid DNA template strand 5′ 3′ 3′
  • 14. Cracking the Code • All 64 codons were deciphered by the mid-1960s • The genetic code is redundant but not ambiguous; no codon specifies more than one amino acid • Codons must be read in the correct reading frame (correct groupings) in order for the specified polypeptide to be produced
  • 15. Figure 17.5 Second mRNA base ThirdmRNAbase(3′endofcodon) FirstmRNAbase(5′endofcodon) UUU UUC UUA UUG Phe Leu Leu Ile Val CUU CUC CUA CUG AUU AUC AUA AUG GUU GUC GUA GUG UCU UCC UCA UCG CCU CCC CCA CCG ACU ACC ACA ACG GCU GCC GCA GCG GAG GAA GAC GAU AAG AAA AAC AAU CAG CAA CAC CAU Ser Pro Thr Ala Glu Asp Lys Asn Gln His Tyr Cys Trp Arg Ser Arg Gly GGG GGA GGC GGU AGG AGA AGC AGU CGG CGA CGC CGU UGG UGA UGC UGUUAU UAC UAA UAG Stop Stop Stop Met or start U C A G U C A G U C A G U C A G U C A G G A C U
  • 16. Evolution of the Genetic Code • The genetic code is nearly universal, shared by the simplest bacteria to the most complex animals • Genes can be transcribed and translated after being transplanted from one species to another Pig expressing a jellyfish gene (b)Tobacco plant expressing a firefly gene (a)
  • 17. Molecular Components of Transcription • RNA synthesis is catalyzed by RNA polymerase, which pries the DNA strands apart and hooks together the RNA nucleotides • RNA synthesis follows the same base-pairing rules as DNA, except uracil substitutes for thymine • The DNA sequence where RNA polymerase attaches is called the promoter; in prokaryotes, the sequence signaling the end of transcription is called the terminator • The stretch of DNA that is transcribed is called a transcription unit
  • 18. Synthesis of an RNA Transcript • The three stages of transcription: – Initiation – Elongation – Termination
  • 19. Figure 17.7-3 Promoter Transcription unit RNA polymerase Start point 1 Template strand of DNARNA transcript Unwound DNA Rewound DNA RNA transcript Direction of transcription (“downstream”) Completed RNA transcript Initiation Elongation Termination 2 3 5′ 3′ 5′ 5′ 3′ 5′ 3′ 5′ 3′ 5′ 5′ 3′ 3′ 5′ 3′ 3′ 5′ 3′ 5′ 3′
  • 20. Figure 17.9 Nontemplate strand of DNA 5′ 3′ 3′ end 5′ 3′ 5′ Direction of transcription RNA polymerase Template strand of DNA Newly made RNA RNA nucleotides A A A A A AA A A C C G G T T T C C CU U C CT T C A T T G G U
  • 21. RNA Polymerase Binding and Initiation of Transcription • Promoters signal the initiation of RNA synthesis • Transcription factors mediate the binding of RNA polymerase and initiation of transcription in eukaryotes • The completed assembly of transcription factors and RNA polymerase II bound to a promoter is called a transcription initiation complex • A promoter called a TATA box is crucial in forming the initiation complex in eukaryotes
  • 22. Promoter Nontemplate strand 15′ 3′ 5′ 3′ Start point RNA polymerase II Template strand TATA box Transcription factors DNA 3′ 5′ 3′ 5′ 3′ 5′ 2 3 Transcription factors RNA transcript Transcription initiation complex 3′5′ 5′ 3′ A eukaryotic promoter Several transcription factors bind to DNA. Transcription initiation complex forms. T A T AAAA TA AT T T T
  • 23. Elongation of the RNA Strand • As RNA polymerase moves along the DNA, it untwists the double helix, 10 to 20 bases at a time • Transcription progresses at a rate of 40 nucleotides per second in eukaryotes • A gene can be transcribed simultaneously by several RNA polymerases
  • 24. Termination of Transcription • The mechanisms of termination are different in prokaryotes and eukaryotes • In prokaryotes, the polymerase stops transcription at the end of the terminator • In eukaryotes, RNA polymerase II transcribes the polyadenylation signal sequence; the RNA transcript is released 10–35 nucleotides past this polyadenylation sequence
  • 25. Concept 17.3: Eukaryotic cells modify RNA after transcription • Enzymes in the eukaryotic nucleus modify pre- mRNA (RNA processing) before the messages are dispatched to the cytoplasm • During RNA processing, both ends of the primary transcript are usually altered (5’ cap and 3’ polyA tail addition) • Also, usually some interior parts of the molecule are cut out, and the other parts spliced together (RNA splicing)
  • 26. Figure 17.10 A modified guanine nucleotide added to the 5′ end Region that includes protein-coding segments 5′ 5′ Cap 5′ UTR Start codon Stop codon G P P P 3′ UTR 3′ AAUAAA AAA AAA Poly-A tail Polyadenylation signal 50–250 adenine nucleotides added to the 3′ end … These modifications share several functions: • They seem to facilitate the export of mRNA • They protect mRNA from hydrolytic enzymes
  • 27. Split Genes and RNA Splicing • Most eukaryotic genes and their RNA transcripts have long noncoding stretches of nucleotides that lie between coding regions • These noncoding regions are called intervening sequences, or introns • The other regions are called exons because they are eventually expressed, usually translated into amino acid sequences • RNA splicing removes introns and joins exons, creating an mRNA molecule with a continuous coding sequence
  • 28. Figure 17.11 Pre-mRNA Intron Intron Introns cut out and exons spliced together Poly-A tail5′ Cap 5′ Cap Poly-A tail 1–30 31–104 105–146 1–146 3′ UTR5′ UTR Coding segment mRNA • In some cases, RNA splicing is carried out by spliceosomes • Spliceosomes consist of a variety of proteins and several small nuclear ribonucleoproteins (snRNPs) that recognize the splice sites
  • 29. Figure 17.12 Spliceosome Small RNAs Exon 2 Cut-out intron Spliceosome components mRNA Exon 1 Exon 2 Pre-mRNA Exon 1 Intron 5′ 5′
  • 30. Ribozymes • Ribozymes are catalytic RNA molecules • Here they function as enzymes that can splice RNA • The discovery of ribozymes rendered obsolete the belief that all biological catalysts were proteins
  • 31. • Three properties of RNA enable it to function as an enzyme – It can form a three-dimensional structure because of its ability to base-pair with itself – Some bases in RNA contain functional groups that may participate in catalysis – RNA may hydrogen-bond with other nucleic acid molecules
  • 32. The Functional and Evolutionary Importance of Introns • Some introns contain sequences that may regulate gene expression • Some genes can encode more than one kind of polypeptide, depending on which segments are treated as exons during splicing • This is called alternative RNA splicing • Consequently, the number of different proteins an organism can produce is much greater than its number of genes
  • 33. • Proteins often have a modular architecture consisting of discrete regions called domains • In many cases, different exons code for the different domains in a protein
  • 34. Figure 17.13 Gene DNA Transcription RNA processing Translation Domain 3 Exon 1 Exon 3Exon 2 IntronIntron Domain 2 Domain 1 Polypeptide
  • 35. Molecular Components of Translation • A cell translates an mRNA message into protein with the help of transfer RNA (tRNA) & ribosomes • tRNAs transfer amino acids to the growing polypeptide in a ribosome • Molecules of tRNA are not identical: – Each carries a specific amino acid on one end – Each has an anticodon on the other end; the anticodon base-pairs with a complementary codon on mRNA
  • 36. Figure 17.14 Polypeptide Amino acids tRNA with amino acid attached Ribosome tRNA Anticodon Codons mRNA 5′ U U U UG G G G C A C C A A A C C G Phe Trp 3′ Gly
  • 37. The Structure and Function of Transfer RNA • A tRNA molecule consists of a single RNA strand that is only about 80 nucleotides long • Flattened into one plane to reveal its base pairing, a tRNA molecule looks like a cloverleaf • Amino acid at one end of sock and anticodon at the other • Because of hydrogen bonds, tRNA actually twists and folds into a three-dimensional molecule • tRNA is roughly L-shaped A C C
  • 38. Figure 17.15 Amino acid attachment site Amino acid attachment site 5′ 3′ A C C A C G C U U A G G C G A U U U A A GAA CC CU * ** C G G U UG C * * * *C CU A G G G G A GAGC C C *U * G A G G U * * * A A A G C U G A A Hydrogen bonds Hydrogen bonds Anticodon Anticodon Symbol used in this book Three-dimensional structure (a) Two-dimensional structure (b) (c) Anticodon 5′3′ A A G 3′ 5′
  • 39. • Accurate translation requires two steps: – First step: a correct match between a tRNA and an amino acid, done by the enzyme aminoacyl-tRNA synthetase – Second step: a correct match between the tRNA anticodon and an mRNA codon Flexible pairing at the third base of a codon is called wobble and allows some tRNAs to bind to more than one codon
  • 40. Figure 17.16-3 Aminoacyl-tRNA synthetase 1 Amino acid and tRNA enter active site. Tyr-tRNA Tyrosine (Tyr) (amino acid) Tyrosyl-tRNA synthetase Complementary tRNA anticodon A AU tRNA Amino acid Using ATP, synthetase catalyzes covalent bonding. 23 ATP AMP + 2 P i Aminoacyl tRNA released. Computer model
  • 41. Ribosomes • Ribosomes facilitate specific coupling of tRNA anticodons with mRNA codons in protein synthesis • The two ribosomal subunits (large and small) are made of proteins and ribosomal RNA (rRNA)
  • 42. Figure 17.17 Exit tunnel Large subunit Small subunit mRNA 3′ 5′ E P A tRNA molecules Growing polypeptide (a) Computer model of functioning ribosome Growing polypeptide Next amino acid to be added to polypeptide chain tRNA 3′ 5′ mRNA Amino end Codons E (c) Schematic model with mRNA and tRNA(b) Schematic model showing binding sites Small subunit Large subunit Exit tunnel A site (Aminoacyl- tRNA binding site) P site (Peptidyl-tRNA binding site) E site (Exit site) mRNA binding site E P A
  • 43. • A ribosome has three binding sites for tRNA: – The P site holds the tRNA that carries the growing polypeptide chain – The A site holds the tRNA that carries the next amino acid to be added to the chain – The E site is the exit site, where uncharged tRNAs leave the ribosome
  • 44. Building a Polypeptide • The three stages of translation: –Initiation –Elongation –Termination –All 3 stages require protein factors and some require energy in the form of GTP.
  • 45. Ribosome Association and Initiation of Translation • Initiation brings together mRNA, a tRNA with the first amino acid, and the two ribosomal subunits • First, a small ribosomal subunit binds with mRNA and a special initiator tRNA • Then the small subunit moves along the mRNA until it reaches the start codon (AUG) • Proteins called initiation factors bring in the large subunit that completes the translation initiation complex
  • 46. Figure 17.18 1 2 P site 3′ Large ribosomal subunit Translation initiation complex Large ribosomal subunit completes the initiation complex. Small ribosomal subunit binds to mRNA. mRNA binding site Small ribosomal subunit Initiator tRNA Start codon 3′5′ E A 3′ 5′ GTP GDP P i + 3′ 5′ 5′ U A C GA UMet Met mRNA
  • 47. Elongation of the Polypeptide Chain • During elongation, amino acids are added one by one to the C-terminus of the growing chain • Each addition involves proteins called elongation factors and occurs in three steps: codon recognition, peptide bond formation, and translocation • Energy expenditure occurs in the first (docking in A site) and third steps(translocation) • Translation proceeds along the mRNA in a 5 → 3 direction′ ′
  • 49. Amino end of polypeptide Codon recognition 1 3′ 5′ E P A sitesite E P A mRNA GTP P i 2 GDP + E P A Peptide bond formation
  • 50. Amino end of polypeptide Codon recognition 1 3′ 5′ E P A sitesite E P A mRNA GTP P i 23 GTP P i GDP + GDP + Translocation E P A Peptide bond formation E P A Ribosome ready for next aminoacyl tRNA
  • 51. Termination of Translation • Termination occurs when a stop codon in the mRNA reaches the A site of the ribosome • The A site accepts a protein called a release factor • The release factor causes the addition of a water molecule instead of an amino acid • This reaction releases the polypeptide, and the translation assembly comes apart
  • 52. Figure 17.20-3 31 2 Release factor 3′ 5′5′ 3′ Stop codon (UAG, UAA, or UGA) Ribosome reaches a stop codon on mRNA. Release factor promotes hydrolysis. Ribosomal subunits and other components dissociate. Free polypeptide 3′ 5′ 2 GTP 2 GDP + 2 P i
  • 53. Polyribosomes • A number of ribosomes can translate a single mRNA simultaneously, forming a polyribosome • Polyribosomes enable a cell to make many copies of a polypeptide very quickly
  • 54. LE 17-20 Ribosomes mRNA 0.1 µm This micrograph shows a large polyribosome in a prokaryotic cell (TEM). An mRNA molecule is generally translated simultaneously by several ribosomes in clusters called polyribosomes. Incoming ribosomal subunits Growing polypeptides End of mRNA (3′ end) Start of mRNA (5′ end) Polyribosome Completed polypeptides
  • 55. Completing and Targeting the Functional Protein • Often translation is not sufficient to make a functional protein • Polypeptide chains are modified after translation • Completed proteins are targeted to specific sites in the cell
  • 56. Targeting Polypeptides to Specific Locations • Two populations of ribosomes are evident in cells: free ribsomes (in the cytosol) and bound ribosomes (attached to the ER) • Free ribosomes mostly synthesize proteins that function in the cytosol • Bound ribosomes make proteins of the endomembrane system and proteins that are secreted from the cell (rough ER) • Ribosomes are identical and can switch from free to bound
  • 57. • Polypeptide synthesis always begins in the cytosol • Synthesis finishes in the cytosol unless the polypeptide signals the ribosome to attach to the ER • Polypeptides destined for the ER or for secretion are marked by a signal peptide • A signal-recognition particle (SRP) binds to the signal peptide • The SRP brings the signal peptide and its ribosome to the ER
  • 58. Figure 17.21 Protein ER membrane Signal peptide removed SRP receptor protein Translocation complex ER LUMEN CYTOSOL SRP Signal peptide mRNA Ribosome Polypeptide synthesis begins. SRP binds to signal peptide. SRP binds to receptor protein. SRP detaches and polypeptide synthesis resumes. Signal- cleaving enzyme cuts off signal peptide. Completed polypeptide folds into final conformation. 1 2 3 4 5 6
  • 59. • A bacterial cell ensures a streamlined process by coupling transcription and translation • In this case the newly made protein can quickly diffuse to its site of function
  • 60. RNA polymerase Polyribosome DNA mRNA 0.25 µm Ribosome DNA mRNA (5′ end) Polyribosome Polypeptide (amino end) Direction of transcriptionRNA polymerase
  • 61. • In eukaryotes, the nuclear envelop separates the processes of transcription and translation • RNA undergoes processes before leaving the nucleus
  • 62. DNA RNA polymerase RNA transcript (pre-mRNA) Intron Exon Aminoacyl-tRNA synthetase Amino acid tRNA Aminoacyl (charged) tRNA mRNA CYTOPLASM NUCLEUS RNA transcript 3′ 5′ Poly-A 5′ Cap 5′ C ap TRANSCRIPTION RNA PROCESSING Poly-A Poly-A AMINO ACID ACTIVATION TRANSLATION Ribosomal subunits E P A E A A C C U U U U U GGG A A A Anticodon Codon Ribosome C A U 3′ A
  • 63. Concept 17.5: RNA plays multiple roles in the cell: a review Type of RNA Functions Messenger RNA (mRNA) Carries information specifying amino acid sequences of proteins from DNA to ribosomes Transfer RNA (tRNA) Serves as adapter molecule in protein synthesis; translates mRNA codons into amino acids Ribosomal RNA (rRNA) Plays catalytic (ribozyme) roles and structural roles in ribosomes
  • 64. Type of RNA Functions Primary transcript Serves as a precursor to mRNA, rRNA, or tRNA, before being processed by splicing or cleavage Small nuclear RNA (snRNA) Plays structural and catalytic roles in spliceosomes SRP RNA Is a component of the the signal- recognition particle (SRP) Video: protein synthesis
  • 65. Concept 17.7: Point mutations can affect protein structure and function • Mutations are changes in the genetic material of a cell or virus • Point mutations are chemical changes in just one base pair of a gene • The change of a single nucleotide in a DNA template strand leads to production of an abnormal protein
  • 66. Figure 17.25 Wild-type β-globin Sickle-cell β-globin Mutant β-globin DNAWild-type β-globin DNA mRNA mRNA Normal hemoglobin Sickle-cell hemoglobin Val U GG A CC G GT Glu G GA G GA C CT3′ 5′ 5′ 5′ 5′ 5′ 3′ 3′ 3′ 3′ 3′ 5′
  • 67. Types of Point Mutations • Point mutations within a gene can be divided into two general categories – Base-pair substitutions – Base-pair insertions or deletions
  • 68. Substitutions • A base-pair substitution replaces one nucleotide and its partner with another pair of nucleotides • Base-pair substitution can cause missense or nonsense mutations • Missense mutations still code for an amino acid, but not necessarily the right amino acid • Nonsense mutations change an amino acid codon into a stop codon, nearly always leading to a nonfunctional protein • Missense mutations are more common
  • 69. Wild type Stop 5′ 3′ 3′ Stop 3′ 5′ 5′ Met (b) Nucleotide-pair insertion or deletion Carboxyl endAmino end mRNA Protein 3′ 5′ 5′ DNA template strand (a) Nucleotide-pair substitution A instead of G U instead of C 5′ 3′ 3′ Stop Extra A Extra U Frameshift (1 nucleotide-pair insertion) missing missing Frameshift (1 nucleotide-pair deletion) missing 3′ 5′ 5′ missing 3′ 5′ 3′ Met Lys Leu Ala 3′ 5′ 3′ T A C T T C A A A C C G A T TA TA G T A A A AC TTTTG G G G G C U A AAAA U U UUUGG A U G A A G G G C UUU U A A AATCGGTTT GG A AA T A C T T C A A C C G A T T A Met Lys Phe Gly T A C C C C GA A A A A AAAA A U G G G G G T T T T TTTTT C G G G C UUUUAA A A 3′ 5′ 5′ T A A A A A T T T TA A A A AACCCC T T T T TTGGGG G G G GUUU U A AA A AU Met Lys Phe Gly Stop U Silent 3′ 5′ 5′ 5′ 3′ 3′ Stop Stop Stop 3 nucleotide-pair deletionNonsense A instead of T U instead of A 5′ 5′ 5′ 3′ 3′ 3′ Met Lys Phe Ser Met Met Phe Gly A T A A T A A T A T instead of C A instead of G Missense U G G G G A AU U U U UU A A A A T T TA A A A AGCCCC A T T T T T TCGGGG 3′ 5′ 5′ 5′ 3′ 3′ T T A A AA A A G U U U U UCGGG T T T T T AGCC CGGG C CTT AAAA T T T TT A AA AA C C C GA A A A ATTTT GGGT C A A AAUCGGGU U U U A
  • 70. Mutagens • Spontaneous mutations can occur during DNA replication, recombination, or repair • Mutagens are physical or chemical agents that can cause mutations
  • 71. 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
  • 72. LE 17-26 TRANSCRIPTION RNA PROCESSING RNA transcript 5′ Exon NUCLEUS FORMATION OF INITIATION COMPLEX CYTOPLASM 3′ DNA RNA polymerase RNA transcript (pre-mRNA) Intron Aminoacyl-tRNA synthetase Amino acid tRNA AMINO ACID ACTIVATION 3′ mRNA A P E Ribosomal subunits 5′ Growing polypeptide E A Activated amino acid Anticodon TRANSLATION Codon Ribosome Review compartments for eukaryotes

Editor's Notes

  1. Don’t make enzyme the substrate accumulates
  2. Auxotrophic
  3. Figure 17.2 Inquiry: Do individual genes specify the enzymes that function in a biochemical pathway? -A mutation in a biochemical pathway can be rescued by downward intermediates Supported the one gene one enzyme hypotheses (logic problem)
  4. Prokaryotic Cells Transcription and translation are coupled because there is no nuclear envelope In eukaryotic cells transcription occurs in nucleus Translation occurs in cytoplasm after being processed in three steps Cap Tail Splicing introns
  5. Figure 17.3 Overview: the roles of transcription and translation in the flow of genetic information
  6. 4 nucleotide bases 20 amino acids 3 base pair sequence is called a codon 64 different codons More than one codon for each amino acid, the codons are degenerates
  7. No comas no overlapping and each of those triplets corresponds to an amino acid when we get to translation
  8. Template strand is complimentary to mRNA In the RNA T is replaced with U Replication and translation occur in the 5’ to 3’ direction
  9. Figure 17.4 The triplet code Amino terminal end is lined up with 5’ end Carboxylic terminal end is lined up with the 3’ end Reading frame consists of three codons and is the one we use for decoding
  10. Figure 17.5 The codon table for mRNA Code is degenerate and universal Start reading codons in AUG -AUG methylamine; not in all of them because they get cleaved *Prokaryotic cells Fmet the beginning *eukaryotic cells: met A double stranded molecule, mRNA, and polypeptide DNA, RNA, and polypeptide sequence
  11. Transgenic means a gene from another organisms is placed in another organism during embryology
  12. RNA polymerase works from 5’ to 3’ -initiation step: when it hits the promoter sequence it binds more tightly to induce torsional stress so a little denatures for mRNA -elongation step -termination step At the end of a gene, the terminator is a stem loop structure, and that stem loop causes mRNA to dissociate
  13. Figure 17.7-3 The stages of transcription: initiation, elongation, and termination (step 3) Elongation: 5’ to 3’ direction as it moves down template syntheszing DNA polymerase Downstream: direction of transcription with positive numbers Termination sequence is in the RNA itself
  14. Figure 17.9 Transcription elongation -10; –35 in prokaryotic cells *
  15. Figure 17.8 The initiation of transcription at a eukaryotic promoter
  16. Figure 17.10 RNA processing: Addition of the 5' cap and poly-A tail
  17. Figure 17.11 RNA processing: RNA splicing
  18. Little proteins and RNA Recognize a specific sequence in the intron In eukaryotic cells translation makes a premessenger rna that
  19. Splicosome is called a ribozyme
  20. More than 70% of genes can code for more than one protein -if a gene has 3 exons you can make a protein by exon 1 and 2 or 1 and 3 or 2 and 3 which gives us a level of complexity that makes us
  21. Protein has different domains: different regions with different purposes
  22. Figure 17.13 Correspondence between exons and protein domains Exon Shuffling can get really complex Signal Transduction Receptor—3 exons Ligand binding domain Dimerization Domain Intracellular Domain that activates the next molecule in signal transduction pathway
  23. Decoding messenger rna and making corresponding amino acid and polypeptide sequence Needs Happens on ribosome Ribosome reads or decodes messenge on messege rna Transfer rna transfer the next amino acid and the catalytic ribosome in the ribosome Transfer rna molecules carry a different amino acid -different structure -Charged: if it carries an amino acid even if it has an uncharged amino acid Anticodon has to be in the opposite direction
  24. Figure 17.14 Translation: the basic concept Looks like worksheet 1.
  25. Figure 17.15 The structure of transfer RNA (tRNA)
  26. Figure 17.16-3 Aminoacyl-tRNA synthetases provide specificity in joining amino acids to their tRNAs (step 3) ATP is used to synthesize the charged RNA
  27. Figure 17.17 The anatomy of a functioning ribosome Ribosomes self assemble in a test tube if added in the right order
  28. Figure 17.18 The initiation of translation
  29. Figure 17.19-1 The elongation cycle of translation (step 1)
  30. Figure 17.19-2 The elongation cycle of translation (step 2)
  31. Figure 17.19-3 The elongation cycle of translation (step 3)
  32. Figure 17.20-3 The termination of translation (step 3)
  33. Figure 17.21 The signal mechanism for targeting proteins to the ER
  34. Figure 17.23 Coupled transcription and translation in bacteria
  35. Figure 17.24 A summary of transcription and translation in a eukaryotic cell
  36. Figure 17.25 The molecular basis of sickle-cell disease: a point mutation One little base pair change
  37. Inserted a base into a reading frame you would change the reading frame down the stream
  38. Figure 17.26 Types of small-scale mutations that affect mRNA sequence Amino acid change doesn't’t change the amino acid protein -
  39. Codes for something that has a job