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Sample Pathway for Control of
Promotor (RNA Tryptophan (trp) Expression
polymerase binding
site)

Enzyme 1
Gene 1

Enzyme 2
Gene 2

Enzyme 3

DNA

Gene 4

not a s na t
i l
r

not pr cs na t
i i
r

Gene 3

Gene 5

mRNA
(transcription unit)

Enzyme 4

Each enzyme
catalyzes the
next series of
reactions
necessary for
tryptophan
production

Enzyme 5

trp

2 Negative Feedback
shut-off systems
Basic Vocabulary
mRNA Transcription Unit:
- grouping of mRNA genes with related
function
- 2 advantages ~
1. organization
2. easy regulation – one “on/off” switch
can be used to control an entire cluster
of related genes
Basic Vocabulary
Operon:
- an entire gene-regulating system in
PROKARYOTES, which includes the
operator, promotor, and all corresponding
genes
Basic Vocabulary

Let’s take a closer
look at the
tryptophan (trp)
operon…an
example of a
“repressor” system

Operator:
- “on-off” switch to genes
- controls the access of RNA polymerase
to genes
- found within the promotor region or
between the promoter and gene-encoding
regions
Overview controls the Trp Operon
Trp operonof Repressor production
Components
of the amino acid tryptophan
r. Trp
promotor
Repressor
5 Genes needed for
Gene
tryptophan production
rp. RNA Pol. o. Operator
(always
binding site
“on”)
r.
rp.
o.
Gene 1 Gene 1Gene 3 Gene 4 Gene 5
RNA polymerase
mRNA transcription unit
mRNA for trp
repressor
Enz 1
Trp repressor protein
translated in its
“inactive” form

Enz 2

Enz 3

Enz 4

Enz 5

trp
How is the trp operon turned off
once enough trp is made?
Trp repressor
gene

RNA pol
binding site

operator

trp

trp

trp
trp

Inactive trp
repressor
protein

trp
Increased concentration of trp (corepressor) increases chances of trp
binding to allosteric site of inactive trp
repressor
How is the trp operon turned off
once enough trp is made?
RNA polymerse,
therefore is
physically blocked
from transcribing
genes for trp

Active trp repressor
can now bind to
operator
Why is this a repressor system?
The trp operon is a repressor operon, meaning
gene expression of the operon is repressed
by the presence of the co-repressor,
tryptophan.
Watch this animation to fully
understand and review this operon,
then take the on-line quiz together
as a class

http://bcs.whfreeman.com/thelifewire/content/chp
Overview of Inducible Lac Operon
The Lac operon controls the production of
the ß-galactosidase, an enzyme that
catalyzes the hydrolysis (break-down) of
lactose into glucose and galactose.
This is an inducible operon, meaning gene
expression ß-galactosidase is stimulated
by the presence of an co-inducer, lactose.
Inducible Lac operon
promotor

I. Lac
Inducer
I.

mRNA for
inducer
protein
inducer
protein in
“active”
form

crp: helps
RNA pol.
to bind

rp: RNA
pol.binding
site

crp.

rp.

o. operator

Genes that
promote ßgalactosidase
production

o.

RNA pol.
blocked from ßgalactosidase
transcription
without lactose
co-inducer
TWO regulatory mechanisms used
to turn on lac operon
1) Presence of lactose as a co-inducer
** reason?
ß-galactosidase is not needed unless lactose
needs to be broken down
2) Low amounts of glucose
** reason?
Recall that lactose breaks down into glucose and
galactose. Low glucose levels signals the cell
for more lactose to be broken down.
Both conditions must be met for the lac operon to turn on.
TWO regulatory mechanisms used
to turn on lac operon
RNA polymerase can bind
only with the help of CRP
transcription factor

RNA pol.

AlloLactose

cAMP

CRP Transcription
Factor
2. cAMP attaches to CRP
transcription factor when
glucose is scarce

1) Allolactose coinducer attaches to
allosteric site of induce
protein, inactivating it
Lac Operon Animation
Watch this animation to fully understand and
review the lac operon!

http://highered.mcgraw-hill.com/olc/dl/120080/bi
How are genes controlled in
eukaryotes?
1) Regulation of Chromatin Structure
2) Pre and post Transcriptional Regulation
3) Pre and Post Translational Regulation
Regulation of Chromatin Structure

N-terminus (amino group) of histone proteins face outwards
from nucleosome
Tails are thus able to be modified chemically
Regulation of Chromatin Structure

Histone Acetylation – neutralizes (+) charges on
•• Methylation  Promotes condensation
tails, which prevents binding to condensation, if
• Phosphorylation  can prevent adjacent
nucleosome  loose chromatin structure
phosphorylation is adjacent to methyl group
results, allowing for increased transcription
Pre-Transcriptional Regulation
• Similar to methods used in bacterial
operons, using proteins that inhibit or
promote binding of RNA pol.
– Distal and Proximal Control Elements
– Proteins involved include:
• Transcription factors
• Activators
• Mediator Proteins
Post-Transcriptional Regulation
RNA Processing
– differential/alternative splicing can produce
different 20 mRNA transcript
Differential splicing redefines
which RNA segments are
considered introns and which
are exons
Post-Transcriptional Regulation
• Time of mRNA degradation can vary
Pre-Translational Regulation
Initiation of Translation
- can be blocked by regulatory proteins
that prevent ribosome binding
- shortened polyA tails in mRNA prevents
translation (polyA tails can be added
during appropriate time)
- global regulatory control of all mRNAs in
cell
Post-Translational Regulation
• During protein processing, folding
• Timing of protein degredation can vary
• Proteasomes degrade proteins that are
tagged by ubiquitine molecules

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Lac Operon

  • 1. Sample Pathway for Control of Promotor (RNA Tryptophan (trp) Expression polymerase binding site) Enzyme 1 Gene 1 Enzyme 2 Gene 2 Enzyme 3 DNA Gene 4 not a s na t i l r not pr cs na t i i r Gene 3 Gene 5 mRNA (transcription unit) Enzyme 4 Each enzyme catalyzes the next series of reactions necessary for tryptophan production Enzyme 5 trp 2 Negative Feedback shut-off systems
  • 2. Basic Vocabulary mRNA Transcription Unit: - grouping of mRNA genes with related function - 2 advantages ~ 1. organization 2. easy regulation – one “on/off” switch can be used to control an entire cluster of related genes
  • 3. Basic Vocabulary Operon: - an entire gene-regulating system in PROKARYOTES, which includes the operator, promotor, and all corresponding genes
  • 4. Basic Vocabulary Let’s take a closer look at the tryptophan (trp) operon…an example of a “repressor” system Operator: - “on-off” switch to genes - controls the access of RNA polymerase to genes - found within the promotor region or between the promoter and gene-encoding regions
  • 5. Overview controls the Trp Operon Trp operonof Repressor production Components of the amino acid tryptophan r. Trp promotor Repressor 5 Genes needed for Gene tryptophan production rp. RNA Pol. o. Operator (always binding site “on”) r. rp. o. Gene 1 Gene 1Gene 3 Gene 4 Gene 5 RNA polymerase mRNA transcription unit mRNA for trp repressor Enz 1 Trp repressor protein translated in its “inactive” form Enz 2 Enz 3 Enz 4 Enz 5 trp
  • 6. How is the trp operon turned off once enough trp is made? Trp repressor gene RNA pol binding site operator trp trp trp trp Inactive trp repressor protein trp Increased concentration of trp (corepressor) increases chances of trp binding to allosteric site of inactive trp repressor
  • 7. How is the trp operon turned off once enough trp is made? RNA polymerse, therefore is physically blocked from transcribing genes for trp Active trp repressor can now bind to operator
  • 8. Why is this a repressor system? The trp operon is a repressor operon, meaning gene expression of the operon is repressed by the presence of the co-repressor, tryptophan.
  • 9. Watch this animation to fully understand and review this operon, then take the on-line quiz together as a class http://bcs.whfreeman.com/thelifewire/content/chp
  • 10. Overview of Inducible Lac Operon The Lac operon controls the production of the ß-galactosidase, an enzyme that catalyzes the hydrolysis (break-down) of lactose into glucose and galactose. This is an inducible operon, meaning gene expression ß-galactosidase is stimulated by the presence of an co-inducer, lactose.
  • 11. Inducible Lac operon promotor I. Lac Inducer I. mRNA for inducer protein inducer protein in “active” form crp: helps RNA pol. to bind rp: RNA pol.binding site crp. rp. o. operator Genes that promote ßgalactosidase production o. RNA pol. blocked from ßgalactosidase transcription without lactose co-inducer
  • 12. TWO regulatory mechanisms used to turn on lac operon 1) Presence of lactose as a co-inducer ** reason? ß-galactosidase is not needed unless lactose needs to be broken down 2) Low amounts of glucose ** reason? Recall that lactose breaks down into glucose and galactose. Low glucose levels signals the cell for more lactose to be broken down. Both conditions must be met for the lac operon to turn on.
  • 13. TWO regulatory mechanisms used to turn on lac operon RNA polymerase can bind only with the help of CRP transcription factor RNA pol. AlloLactose cAMP CRP Transcription Factor 2. cAMP attaches to CRP transcription factor when glucose is scarce 1) Allolactose coinducer attaches to allosteric site of induce protein, inactivating it
  • 14. Lac Operon Animation Watch this animation to fully understand and review the lac operon! http://highered.mcgraw-hill.com/olc/dl/120080/bi
  • 15. How are genes controlled in eukaryotes? 1) Regulation of Chromatin Structure 2) Pre and post Transcriptional Regulation 3) Pre and Post Translational Regulation
  • 16. Regulation of Chromatin Structure N-terminus (amino group) of histone proteins face outwards from nucleosome Tails are thus able to be modified chemically
  • 17. Regulation of Chromatin Structure Histone Acetylation – neutralizes (+) charges on •• Methylation  Promotes condensation tails, which prevents binding to condensation, if • Phosphorylation  can prevent adjacent nucleosome  loose chromatin structure phosphorylation is adjacent to methyl group results, allowing for increased transcription
  • 18. Pre-Transcriptional Regulation • Similar to methods used in bacterial operons, using proteins that inhibit or promote binding of RNA pol. – Distal and Proximal Control Elements – Proteins involved include: • Transcription factors • Activators • Mediator Proteins
  • 19.
  • 20. Post-Transcriptional Regulation RNA Processing – differential/alternative splicing can produce different 20 mRNA transcript Differential splicing redefines which RNA segments are considered introns and which are exons
  • 21. Post-Transcriptional Regulation • Time of mRNA degradation can vary
  • 22. Pre-Translational Regulation Initiation of Translation - can be blocked by regulatory proteins that prevent ribosome binding - shortened polyA tails in mRNA prevents translation (polyA tails can be added during appropriate time) - global regulatory control of all mRNAs in cell
  • 23. Post-Translational Regulation • During protein processing, folding • Timing of protein degredation can vary • Proteasomes degrade proteins that are tagged by ubiquitine molecules