Control of Mechanism

of
Gene Expression

Dr. Tapan Kr. Dutta

Panskura Banamali College
 All Genes Cant be Expressed
At The Same Time
Some genes are needed for the function of all
cells all the time. These genes are called
constitutive genes and are expressed by all cells.
Other genes are only needed by certain cells or at
specific times. The expression of these inducible
genes is tightly controlled in most cells.
For example, beta cells in the pancreas make the
protein insulin by expressing the insulin gene. If
neurons expressed insulin, problems would result.
Operons Are Groups Of Genes
Expressed By Prokaryotes

The genes grouped in an operon are all

needed to complete a given task
Each operon is controlled by a single
control sequence in the DNA
Because the genes are grouped together,
they can be transcribed together then
translated together
 The Lac Operon
Genes in the lac operon allow E. coli bacteria to
metabolize lactose
Lactose is a sugar that E. coli is unlikely to
encounter, so it would be wasteful to produce the
proteins needed to metabolize it unless necessary
Metabolizing lactose for energy only makes sense
when two criteria are met:
Other more readily metabolized sugar (glucose) is
unavailable
Lactose is available
 The Lac Operon - Parts
The lac operon is made up of a control region and
four genes
The four genes are:
LacZ - b-galactosidase - An enzyme that hydrolizes
the bond between galactose and glucose
LacY - Codes for a permease that lets lactose across
the cell membrane
LacA - Transacetylase - An enzyme whose function in
lactose metabolism is uncertain
Repressor - A protien that works with the control
region to control expression of the operon
 The Lac Operon - Control
The control region is made up of two parts:
Promoter
These are specific DNA sequences to which RNA
Polymerase binds so that transcription can occur
The lac operon promoter also has a binding site for
another protein called CAP
Operator
The binding site of the repressor protein
The operator is located down stream (in the 3
direction) from the promoter so that if repressor is
bound RNA Polymerase cant transcribe
 The Lac Operon:
When Glucose Is Present But Not Lactose
Hey man, Im Come on,
constitutive let me through

CAP RNA
Repressor Binding Promoter Operator LacZ LacY LacA
Pol. Repressor

Repressor No way
mRNA
Jose!

Repressor
CAP
 The Lac Operon:
When Glucose And Lactose Are Present
Hey man, Im Great, I can
constitutive transcribe!

CAP RNA RNA

Repressor Binding Promoter Operator LacZ LacY LacA
Pol. Pol.

Repressor X Repressor
mRNA
This lactose has
Repressor CAP
bent me
out of shape

Repressor

Some transcription
occurs, but at a slow rate
 The Lac Operon:
When Lactose Is Present But Not Glucose
Bind to me
Hey man, Im Yipee!
Polymerase
constitutive

CAP RNA RNA

Repressor Binding Promoter Operator LacZ LacY LacA
CAP Pol. Pol.
cAMP

Repressor X Repressor
mRNA CAP
cAMP

Repressor

This lactose has

bent me
Repressor cAMP CAP
out of shape
 The Lac Operon:
When Neither Lactose Nor Glucose Is Present
Alright, Im off to
Bind to me the races . . .
Hey man, Im Come on, let
Polymerase
constitutive me through!

CAP RNA
Repressor Binding Promoter Operator LacZ LacY LacA
CAP Pol. Repressor
cAMP

Repressor STOP
mRNA Right there CAP
Polymerase cAMP

Repressor cAMP CAP

 The Trp Operon
Genes in the trp operon allow E. coli bacteria
to make the amino acid tryptophan
Enzymes encoded by genes in the trp operon
are all involved in the biochemical pathway that
converts the precursor chorismate to
tryptophan.
The trp operon is controlled in two ways:
Using a repressor that works in exactly the opposite
way from the lac operon repressor
Using a special attenuator sequence
 The Tryptophan
COO-
Biochemical Pathway 5-Phosphoribosyl-
Glutamine Glutamate + PPi -OOC
Pyruvate COO- a-Pyrophosphate
CH2 NH2
Anthranilate synthetase HN
O C COO- -2O P O CH2
3 O
H Anthranilate synthetase
H H N-(5-
Chorismate (trpE and D) Antrhanilate
HO H H H Phosphoribosyl)
N-(5-Phosphoribosyl)-anthranilate OH
-anthranilate
OH
isomerase Indole-3-glycerol
OH OH phosphate synthetase (trpC) OH OH
-OOC CO2+H2O
-2O PO CH2 C C C OH
-2O PO
3 CH2 C C C
3 N-(5-Phosphoribosyl)-
Enol-1-o- H H C Anthranilate isomerase Indole- H H C
Carboxyphenylamino H H
N 3-glycerol phosphate synthetase H N
H
-1-deoxyribulose phosphate Glyceraldehyde- Tryptophan synthetase Indole-3-glycerol phosphate
(trpB and A) H
3-phosphate
-OOC C CH2
Serine H2O
NH3+
Tryptophan synthetase
N
H N
Indole H Tryptophan
 The Trp Operon:
When Tryptophan Is Present
Hey man, Im Foiled
constitutive
Again!

RNA
Repressor Promo. Operator Lead. Aten. trpE trpD trpC trpB trpA
Pol. Repressor
Trp

Repressor
mRNA STOP
Right there
Polymerase

Trp
Repressor
 Attenuation
The trp operon is controlled both by a
repressor and attenuation
Attenuation is a mechanism that works only
because of the way transcription and
translation are coupled in prokaryotes
Therefore, to understand attenuation, it is
first necessary to understand transcription
and translation in prokaryotes
 Transcription And Translation
In Prokaryotes
5 3

3 5
RNA
Pol.

Ribosome

mRNA
Ribosome
5
 The Trp Leader and
Attenuator
Met-Lys-Ala-Ile-Phe-Val-
AAGUUCACGUAAAAAGGGUAUCGACA-AUG-AAA-GCA-AUU-UUC-GUA-

Leu-Lys-Gly-Trp-Trp-Arg-Thr-Ser-STOP
CUG-AAA-GGU-UGG-UGG-CGC-ACU-UCC-UGA-AACGGGCAGUGUAUU

1 2
CACCAUGCGUAAAGCAAUCAGAUACCCAGCCCGCCUAAUGAGCGGGCUUUU

3 4
Met-Gln-Thr-Gln-Lys-Pro
UUUU-GAACAAAAUUAGAGAAUAACA-AUG-CAA-ACA-CAA-AAA-CCG
trpE . . .
Terminator
The mRNA Sequence Can
Fold In Two Ways

1 2
1 2
3 3
4
4
Terminator
haripin
 The Attenuator
When Starved For Tryptophan
5 3

3
5
Help, RNA
I need Pol.
Tryptophan

2
Ribosome 3
4
1
 The Attenuator
When Tryptophan Is Present

5 3

3 Ribosome 5

RNA
2 Pol.
3
4
1
 Control Of Expression In
Eukaryotes
Some of the general methods used to control
expression in prokaryotes are used in eukaryotes,
but nothing resembling operons is known
Eukaryotic genes are controlled individually and
each gene has specific control sequences
preceding the transcription start site
In addition to controling transcription, there are
additional ways in which expression can be
controlled in eukaryotes
 Eukaryotes Have Large
Complex Geneomes
The human genome is about 3 x 109 base
pairs or 1 m of DNA
Because humans are diploid, each nucleus
contains 6 3 x 109 base pairs or 2 m of
DNA
That is a lot to pack into a little nucleus!
 Eukaryotic DNA Must be
Packaged
Eukaryotic DNA exhibits many levels of
packaging
The fundamental unit is the nucleosome,
DNA wound around histone proteins
Nucleosomes arrange themselves together
to form higher and higher levels of
packaging.
Highly Packaged DNA Cannot
be Expressed
The most highly packaged form of DNA is
heterochromatin
Heterochromatin cannot be transcribed,
therefore expression of genes is prevented
Chromosome puffs on some insect
chomosomes illustrate where active gene
expression is going on
 Only a Subset of Genes is
Expressed at any Given Time
Ittakes lots of energy to express genes
Thus it would be wasteful to express all
genes all the time
By differential expression of genes, cells
can respond to changes in the environment
Differential expression, allows cells to
specialize in multicelled organisms.
Differential expression also allows
organisms to develop over time.
Control of Gene Expression

Cytoplasm
Packaging
Degradation
DNA

Transcription
RNA
Transportation
RNA Modification
Processing
Degradation etc.
mRNA G AAAAAA G AAAAAA

Export Translation
Nucleus
Logical Expression Control Points
DNA packaging

Increasing cost
Transcription The logical
RNA processing place to
mRNA Export control
mRNA masking/unmasking expression is
and/or modification before the
mRNA degradation gene is
Translation transcribed
Protein modification
Protein transport
Protein degradation
 A Simple Eukaryotic Gene
Transcription
Start Site 3 Untranslated Region
5 Untranslated Region
Introns

5 Exon 1 Int. 1 Exon 2 Int. 2 Exon 3 3

Promoter/ Terminator
Control Region
Exons Sequence
RNA Transcript
 Enhancers
DNA Many bases
5 3
Enhancer Promoter Transcribed Region

5 TF TF 3

5 TF TF RNA RNA 3
Pol. Pol.
5
RNA
 Eukaryotic mRNA
5 Untranslated Region 3 Untranslated Region

5 G Exon 1 Exon 2 Exon 3 AAAAA 3

Protein Coding Region
5 Cap 3 Poly A Tail

RNA processing achieves three things:

Removal of introns
Addition of a 5 cap
Addition of a 3 tail
This signals the mRNA is ready to move out
of the nucleus and may control its life span
in the cytoplasm