BIO112: MOLECULAR

BIOLOGY AND GENETICS

Mr. Derrick Banda MSc, BSc
 LECTURE 5
REGULATION OF GENE
EXPRESSION
PART 2
 THE OPERON MODEL
• Jacob and Monod proposed the operon model in
1961 for the co-ordinate regulation of transcription
of genes involved in specific metabolic pathways.

• An operon is a group of related and closely linked

structural genes and associated control gene which
regulate the metabolic activity.

• In bacterial DNA, a cluster of genes transcribed

from one promoter that gives rise to a
polycistronic mRNA.
 THE OPERON MODEL
• The operon is a unit of gene expression and regulation which typically
made up of four (4) basic DNA components:
1. The regulator gene codes for a repressor protein that binds to the
operator, obstructing the promoter (thus, transcription) of the
structural genes.

2. Promoter – a sequence that enables a gene to be transcribed. The

promoter is recognized by RNA polymerase, which then initiates
transcription.

3. Operator – a segment of DNA that a repressor binds to. It is

classically defined in the lac operon as a segment between the
promoter and the structural genes of the operon.

4. Structural genes – the genes that are co-regulated by the operon.

 THE OPERON MODEL
• The operon is a unit of gene expression and regulation which
typically made up of four (4) basic DNA components:
1. Regulator gene
2. Promoter
3. Operator
4. Structural genes
 THE OPERON AND GENE REGULATION
• One of the first examples of gene regulation to be
worked out genetically was the lac operon of E. coli
by Francois Jacob and Jacques Monod in 1961.

• Regulation of gene expression can be done by some

operon pathways such as;
1.Lac (Lactose) operon.
2.Tryptophan (trp) operon.
 THE INDUCIBLE AND REPRESSIBLE OPERONS
• Prokaryotic operons are commonly controlled
by the binding of repressors to operator
regions, thereby preventing the transcription
of the structural genes. Such operons are
classified as;

1.Inducible operons (lac operon)

2.Repressible operons (trp operon)

 INDUCIBLE AND REPRESSIBLE OPERON SYSTEMS
1. In inducible systems, the substrate of a metabolic
pathway (the inducer) interacts with a regulatory
protein (the repressor), rendering the repressor
incapable of binding to the operator and thus allowing
transcription. A lac operon is said to be an inducible
operon because the presence of lactose induces the
expression of genes.

2. In repressible systems, the product of a metabolic

pathway (the co-repressor) binds to a regulatory
protein, which is then able to bind to the operator and
block transcription. Repressible operons, like the
tryptophan (trp) operon.
 THE lac OPERON MODEL
• Lactose is a disaccharide found in milk or milk
products that consists of two sugars: glucose and
galactose.

• Escherichia coli can use lactose as a source of

carbon. The enzymes required for the use of lactose
as a carbon source are only synthesized when
lactose is available as the sole carbon source.
 THE lac OPERON MODEL
• The lactose operon (or lac operon) consists of three structural
genes: lacZ, which codes for β-galactosidase, an enzyme
responsible for hydrolysis of lactose to galactose and glucose;
lacY, which encodes a galactoside permease which is
responsible for lactose transport across the bacterial cell wall;
and lacA, which encodes a thiogalactoside transacetylase.
THE STRUCTURE OF lac OPERON
• The lac operon consists of:
1. Regulatory gene – It codes for the repressor protein.
2. Promoter
3. Operator
4. Structural genes (Lac Z, Lac Y and Lac A)that are linked in sequence on
one strand of DNA. The Z gene – It codes for galactosidase which
catalyzes the hydrolysis of lactose into glucose and galactose. The Y gene –
It codes for permease which regulates the lactose permeability in the cell.
The A gene – It codes for transacetylase which assists the enzyme
galactosidase.
 THE STRUCTURE OF lac OPERON
• The three structural genes are encoded in a single transcription
unit, lacZYA, which has a single promoter, Plac. This
organization means that the three lactose operon structural
proteins are expressed together as a polycistronic mRNA
containing more than one coding region under the same
regulatory control.
 The lac operon Regulation
• All the lac operon genes help in the lactose metabolism.

• The lac operon can be regulated in two ways:

1. Negative regulation by repressors

2. Positive regulation by activators

• Specific control of the lac genes depends on the

presence or absence of lactose in a cell.
 POSITIVE AND NEGATIVE REGULATION
• Genes can be regulate at the promoter site in two ways:
1. Negative regulation by repressors: In negative
regulation, a repressor binds the operator sequence to
prevent transcription. Only in the absence of repressor
protein can the gene be transcribed.

2. Positive regulation by activators: In positive

regulation, a transcriptional activator protein binds the
operator, to promote transcription. In the absence of
the activator protein, there is no transcription.
 NEGATIVE REGULATION OF LAC OPERON
• Negative regulation involves the binding of a repressor
to the operator to prevent transcription.

• In negative inducible operons, a regulatory repressor

protein is normally bound to the operator, which
prevents the transcription of the genes on the operon

• For the lac (inducible) operon to be expressed, lactose

must be present. This makes sense for the cell because
it would be energetically wasteful to create the
enzymes to process lactose if lactose was not available.
 The Lac Operon as an inducible operon
• The genes required for lactose metabolism and
utilization in E. coli are clustered into a single region
on the chromosome. It is the inducible operon since
the presence of lactose induce the operon to be
switched on.
 ABSENCE OF LACTOSE
• In the absence of lactose, the regulatory gene
transcribes mRNA to synthesize a protein called
Lac repressor protein.
• The lac repressor binds tightly to the Operator. It
gets in RNA polymerase’s way, preventing
Transcription.
 LACTOSE AS AN INDUCER
• The lac operon is a negatively controlled inducible
operon, where the inducer molecule is allolactose.

• When lactose is present, the lactose inside the cell is

converted to allolactose.

• Allolactose binds to the repressor and changing its

shape so that it is no longer able to bind to the operator
DNA.

• Removal of the repressor in the presence of lactose

allows RNA polymerase to move through the operator
region and begin transcription of the lac structural
genes. Thus, lac operon is negatively regulated in this
case.
 LACTOSE AS AN INDUCER
• If an inducer molecule is present, it binds to the
repressor and changes its shape so that it is unable to
bind to the operator. This allows for expression of
the operon.
 How two regulatory protein (CAP and Repressor
protein) control the lac operon
• The lac operon has two regulatory sites and these are the
CAP (Catabolite activator protein) site (Activator
switch) and the Operator site (Repressor switch).
 Catabolite (Glucose) repression
• When glucose is available, gene that
participate in the metabolism of other sugars
are repressed, in a phenomenon known as
catabolite repression.

• Glucose does not bind to a regulatory protein

to negatively regulate other operons.
 Catabolite (Glucose) repression
• Glucose inhibits adenylate cyclase, which
synthesizes signalling molecule cAMP (cyclic
Adenine Mono-phosphate) from ATP.
 CAP (catabolite activator protein)
• There is a protein called CAP (catabolite activator
protein) in E. coli that binds cAMP.
• When CAP is bound to cAMP, it changes shape and
becomes a positive regulator of the lac operon.
 POSITIVE REGULATION
• Catabolite repression is a type of positive regulation in the
lac operon.
• The CAP protein, is an example of positive regulation that
is activated by the presence of a different small effector
molecule (cAMP).

• The CAP-cAMP complex binds to a CAP Binding site on the

DNA, near the promoter and stimulates the binding of RNA
polymerase, so that the lac genes can be transcribed

• A cellular level of signaling molecule cAMP (cyclic Adenine

Mono-phosphate) are controlled by glucose; allolactose
level increases the abundance of cAMP & enhance the
transcription of the lac structural genes.
 Glucose levels regulate cAMP levels
• The concentration of cAMP is inversely
proportional to the abundance of glucose: when
glucose concentrations are low, an enzyme
called adenylate cyclase is able to produce cAMP
from ATP.
Levels of cAMP and rate of Transcription
• The amount of cAMP and the rate of Transcription of
the lac operon are inversely related to the concentration
of Glucose.
 The state of the lac operon under the
three conditions

1. Conditions with glucose only, no lactose

2. Conditions with glucose + lactose

3. Conditions with lactose, no glucose

1. Conditions with Glucose only, No lactose;
• The level of cAMP is low, so CAP is not activated
(absence of positive regulation). There is no lactose, so
the lac repressor remains bound (negative regulation).
Under these conditions there is no transcription of
the lac operon.
2. Conditions with glucose + lactose;
• The level of cAMP is low, so CAP is not activated
(absence of positive regulation). In the presence of
lactose, the lac repressor is no longer bound (absence
of negative regulation). Under these conditions there is
a low level of transcription of the lac operon.
 Lactose present, Glucose present
• When glucose levels are high, there is catabolite repression of
operons encoding enzymes for the metabolism of alternative
substrates.
• Low cAMP levels under these conditions, reduces the amount of the
CAP-cAMP complex to bind to the CAP (CRP) binding site of the
DNA molecule to activate transcription of these operons.
3. Conditions with lactose, no glucose;
• The level of cAMP is high, so CAP is activated
(positive regulation). In the presence of lactose,
the lac repressor is no longer bound (absence of
negative regulation). Under these conditions there is a
very high level of transcription of the lac operon.
 Lactose present, Glucose absent
• When glucose is scarce, the accumulating cAMP caused by increased
adenylyl cyclase activity binds to catabolite activator protein
(CAP), also known as cAMP receptor protein (CRP).

• Binding of the CAP-cAMP complex to CRP binding site increases

the binding ability of RNA polymerase to the promoter region to
initiate the transcription of the lac structural genes.
 End-product Inhibition of the Lac
Operon Mechanism
• When amount of glucose builds up (i.e from lactose
metabolism). The CAP site causes the lac genes not to be
transcribed properly.

• Increasing levels of glucose in the cell leads to lower levels

of signaling molecule cAMP (cyclic Adenine Mono-
phosphate) which binds to the CAP.

• Hence, glucose as an end product may inhibit further

transcription and translation of lac genes, resulting in
reduced metabolism of Lactose (End-product inhibition).
END OF LECTURE!
THANK YOU!