CONTROL OF GENE EXPRESSION IN

BACTERIA

Dr.Minal Trivedi
Principal,
HVHP Institute of Post Graduate Studies and
Research,
Kadi Sarva Vishwavidyalaya
Kadi – Gujarat
 CONTENT
 Control of Gene expression
 Operon model

2
 Types of Operon
 Inducible Operon
 Repressible Operon
 Lac- operon
 Negative Control
 Catabolic repression
 Trp-Operon
LEARNING OUTCOMES

 How Gene expression is controlled in Bacteria

 Understanding of Operon model

 Understanding of Types of Operon

 Mechanism of Lactose Utilization in Bacterial –

Example of Inducible Operon
 Mechanism of tryptophan Biosynthesis in Bacterial –
Example of repressible Operon
REGULATION OF GENE EXPRESSION

 Genomic DNA contains both structural genes, which encode

products that serve as cellular structures or enzymes,
and regulatory genes, which encode products that regulate
gene expression. The expression of a gene is a highly
regulated process.
 OPERON
 Operon- is a co-ordinated group of genes which are all transcribed
together & regulate a metabolic pathway as a unit.
 In 1961, two French scientists ,Francis Jacob and Jacques Monad
at Pasteur Institute of Paris, proposed a mechanism called opeon
model for regulation of gene expression
 Operon is a part of Genetic material or DNA.

 Operon are of two types.

 1. Inducible Operon
 2. Repressible Operon.
OPERON STRUCTURE
 INDUCIBLE OPERON

 Inducible operons are usually "off," but can be turned

"on" by a small molecule.
 The molecule is called an inducer, and the operon is said
to be inducible.
 For example, the lac operon is an inducible operon that
encodes enzymes for metabolism of the sugar lactose.

Active Repressor+ Inducer = Inactive Repressor

INDUCTION

 In induction, synthesis of enzyme takes place in response

to appearance of specific substrate in the medium.

 The phenomenon of induction can be explained by taking

the example of working of Lactose operon.
 REPRESSIBLE OPERON
 An operon that is regulated by a co-repressor (chemical
substance) is called as the repressible operon.
 These operons are usually "on," but can be turned "off"
by a small molecule. The molecule is called a corepressor,
and the operon is said to be repressible. For example,
the trp operon is a repressible operon that encodes
enzymes for synthesis of the amino acid tryptophan.

Inactive Repressor+ Co-repressor = Active Repressor

OPERONS
 An operon is a cluster of coordinately regulated genes. It
contains:
 Structural genes: encode enzymes

 Regulatory genes: encode repressors or activators of expression

 Regulatory sites: e.g. promoters, operators

THE LAC OPERON OF E.COLI
In 1961,Francois Jacob and Jacques Monod
described the “operon model” for the genetic
control of lactose metabolism in E.coli.
This work and subsequent research on the
genetics of lactose metabolism established the
operon as the operon as the basic unit of
transcriptional control in bacteria.
THE LAC OPERON
 The lactose operon designated as lac
operon.
 The lac operon codes for enzymes
involved in the catabolism (degradation)
of lactose.
 lactose is the disaccharide which is
made up of glucose & galactose.
 It is the inducible operon since the
presence of lactose induce the operon to
switched on.
THE LAC OPERON OF E. COLI

A negative inducible operon

 Inducer: allolactose

 lacI: repressor encoding gene

 lacP: operon promoter

 lacO: operon operator

THE LAC OPERON OF E. COLI

• Structural genes

• lacZ: encoding β-galactosidases

• lacY: encoding permease
• lacA: encoding transacetylase

• The repression of the lac operon never

completely shuts down transcription.
DESIGNATION OF GENE CODES FOR ENZYME
FUNCTION OF THE ENZYME
 lac Z :β-galactosidase -Breaks down
lactose into glucose & galactose.
 lac y: galactose permease -This protein,
found in the E.coli cytoplasmic
membrane, actively transports lactose
into the cells
 lac a :Thio-galactoside trans acetylase
-The function of this enzyme is not
known. It is coded for by the gene lacA.
THE LAC OPERON HAS A SECOND LAYER OF
CONTROL: CATABOLITE REPRESSION

 catabolite repression – The ability of glucose to

prevent the expression of a number of genes.
 In bacteria this is a positive control system; in
eukaryotes, it is completely different.

 Catabolite repressor protein (CRP) is an

activator protein that binds to a target sequence
at a promoter.
POSITIVE CONTROL AND CATABOLITE REPRESSION
 This is a positive control mechanism:
 The positive effect is activated by catabolite
activator protein (CAP) also known as cAMP
receptor Protein ( CRP).
 CRP increase the affinity of RNA polymerase
to bind promoter.
 CRP is inactive protein- its activation require
cAMP
 bind with cAMP
Inactive CRP active cAMP-CAP
complex
( Also known as active CRP
act as activator )
CAMP
 Cyclic AMP
 Generated from AMP by activity of enzyme
Adenylate Cyclase
 cAMP – adenosine-3′,5′-cyclic monophosphate

 The concentration of cAMP is inversely proportional

to the level of available glucose.
 cAMP is binded to CAP, together CAP–cAMP
complex binds to a site slightly upstream from the
lac gene promoter.

High Glucose High ATP Less cAMP

Less Glucose Less ATP High cAMP

cAMP converts an activator protein CRP to a form that
binds the promoter and assists RNA polymerase in
initiating transcription.
  A dimer of CRP is activated by
a single molecule of cyclic
AMP (cAMP).
 cAMP is controlled by the level
of glucose in the cell; a low
glucose level allows cAMP to
be made.
 CRP interacts with the C-
terminal domain of the α
subunit of RNA polymerase to
activate it.

By reducing the level of cyclic AMP, glucose

inhibits the transcription of operons that
require CRP activity.
TRYPTOPHAN OPERON
OPERON FOR BIOSYNTHESIS OF AMINO ACIDS
 The 20 common amino acids are required in large
amount for protein synthesis, and E.coli can
synthesize all of them.

 The genes for the enzymes needed to synthesize a

given amino acid are generally clustered in an operon
and are expressed whenever existing supplies of that
amino acid are inadequate for cellular requirements.

 When the amino acid is abundant the biosynthetic

enzymes are not needed and the operon is repressed.
 TRP OPERON IS A REPRESSIBLE OPERON

 trp operon responds to the cellular level of amino acid,

—
here Tryptophan.

 The operon is switched off whenever cellular level of

—
tryptophan is high (repressible).

 The E.coli trp operon is regulated at two levels Blocking

the transcription initiation via repressor-operator
interaction and another is attenuation control.

 The tryptophan operon is the regulation of transcription

of the gene responsible for biosynthesis of tryptophan
from chorismate.
 The tryptophan (trp) operon contains five structural
genes encoding enzymes for tryptophan biosynthesis
with an upstream trp promoter (Ptrp) and trp operator
sequence (Otrp).
 Structural genes are TrpE, TrpD, TrpC, TrpB and
TrpA
 trpE: It enodes the enzyme Anthranilate synthase I

 trpD: It encodes the enzyme Anthranilate synthase II

 trpC: It encodes the enzyme N-5’-Phosphoribosyl

anthranilate isomerase and Indole-3-glycerolphosphate
synthase
 trpB: It encodes the enzyme tryptophan synthase-B sub
unit
 trpA: It encode the enzyme tryptophan synthase-A sub
unit
PRESENCE OF HIGH LEVEL OF TRYPTOPHAN
ABSENCE OF TRYPTOPHAN
 SOME OPERONS REGULATE TRANSCRIPTION THROUGH
ATTENUATION, THE PREMATURE TERMINATION OF
TRANSCRIPTION
• Attenuation is a proposed mechanism of control in
some bacterial operons which results in premature
termination of transcription and is based on the fact that,
in bacteria, transcription and translation proceed
simultaneously.
• Attenuation is a type of gene regulation in to some
bacterial operons in which transcription begins at the
transcription start site but terminate prematurely, before
RNA Polymerase even reaches the structural gene

• Affects the continuation of transcription, not its

initiation.
• This action terminates the transcription before it reaches
the structural genes.
ATTENUATION
 Attenuation is the regulation of bacterial operons by
—
controlling termination of transcription at a site
located before the first structural genes.

 Most of the amino acid bio synthetic pathways are

—
regulated by attenuation.

 The attenuation is based on the ability of RNA to

—
form alternative structures through intra-molecular
base pairing.

 In E.coli, the attenuation requires close link between

—
transcription and translation.
 In bacteria, transcription and translation
occurs simultaneously.

 The translation starts before transcription

completes.

 In this attenuation mechanism, rate of

translation determines whether transcription
continues or terminates. Therefore the
attenuation mechanism is only found in
bacteria but not in eukaryotic cell
ATTENUATION CONTROL OF TRYPTOPHAN
OPERON

 Leader sequence (trpL) play important role in

attenuation.

 Leader sequence contains such a nucleotide sequence

that mRNA transcribed from it contains four specific
region.

 Region 1, region 2, region 3 and Region 4. Region 3 is

complementary to both region 2 and region 4.
 Region 1 is the most important region that
determines whether to form loop between
region 2-region 3 or region 3-region 4.

 The region 1 consists of sequence of 14

codons, out of which two codons are
tryptophan codon (codon 10 and 11).
 If region 3 and region 4 base pair with
each other, they form a loop like
structure called attenuator and it function
as transcriptional termination.

 If pairing occur between region 2 and

region 3, then no such attenuator form so
that transcription continues.
HIGH LEVEL OF TRYPTOPHAN
LOW LEVEL OF TRYPTOPHAN
 REFERENCE :

 Genetics – A Conceptual Approach by Benjamin A Pierce ( 7th

Edition)
 Microbiology by Pelczar MJ, Chan ECS and Krieg NR(1993)