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OPERON SYSTEM
CONTENT
★Operon system and types
★Lac operon
★Trp operon
★Ara operon
Operon System:
• An operon is a functional unit of DNA in
prokaryotes that contains a group of genes
regulated together.
• These genes are transcribed as a single
mRNA molecule, allowing coordinated
expression of proteins involved in related
functions (e.g., metabolic pathways).
Types of Operons:
★Inducible Operon:
o Normally off but can be activated in the
presence of a specific substrate.
o Example: Lac Operon (activated by lactose).
★Repressible Operon:
o Normally on but can be turned off when the
end product is abundant.
o Example: Trp Operon (repressed by
tryptophan).
Lac Operon:
In 1961, François Jacob and Jacques Monod
at the Pasteur Institute in Paris
discovered the lac operon while studying how
bacteria (particularly Escherichia coli)
adapt to metabolize lactose. They were
awarded the Nobel Prize in Physiology or
Medicine (1965).
Codes for enzymes involved in the catabolism
of lactose (disaccharide made up of
Glucose + Galactose).
• It is the inducible operon since the presence
of lactose induce the operon to switch on .
Structure of Lac Operon:
1.Regulatory gene:
o Regulatory gene is the i gene that code for
the repressor protein.
o This i gene is expressed in all the time
hence it is also known as constitutive gene.
2.Structural gene:
o Three structural gene Lac z, Lac y, Lac a
involved in the synthesis of enzymes for
the lactose catabolism.
o The 3 genes as transcribed in a polycistronic
mRNA :
★ lacZ encode beta-galactosidase enzyme
(tetramer) that hydrolyzes lactose into
glucose and galactose. It also catalyzes
isomerization of lactose to allolactose that
acts as an inducer.
★ lacY encode galactoside
permease/M-protein (monomer) that transport
lactose into the
cell.
★lacA encode thiogalactoside transacetylase
enzyme (dimer) that transfers an acetyl group
from acetyl-coA to beta-galactoside. This
enzyme is not essential for lactose
metabolism. It
appears to play a role in the detoxification of
the galactosides that are also imported by the
permease.
3. Promoter : It is a region of DNA/gene
where DNA polymerase binds and initiate
transcription.
4. Operator : It is a region of DNA/gene where
repressor protein (product of regulatory gene
lacI) binds and regulates transcription of
structural genes of lac operon. Lac operon has
three operator sites: one primary/major
operator O1 and two secondary operators
O2 and O3.
★Negative control of Lac-operon:
»The lac repressor is synthesized through the
activity of Lac i gene called the regulator
gene.
» This repressor is an allosteric protein ( a
protein whose shape is changed when it binds
a
particular molecule)
1.It binds the operator site
2.It binds the lactose sugar (inducer)
. In the absence of the Inducer, the DNA
binding site of the repressor is functional. »The
repressor protein binds to the DNA at the
operator site and blocks the transcription of lac
genes by RNA Polymerase.
»The lac enzyme synthesis is inhibited.
Positive control of lac operon:

»it is an additional regulatory mechanism
which allows the lac operon to sense the
presence
of glucose, an alternative and preferred energy
source to lactose (cells will use up the
glucose first).
»The presence of glucose in the cell switches
off the lac operon by a mechanism called
catabolite repression, which involved a protein
called Catabolite Activator Protein
(CAP).
. CAP only binds in the presence of a
derivative of ATP called cyclic Adenosine
Monophosphate (cAMP).
•CAP binds to a DNA sequence upstream of
the lac promoter and enhances binding of the
RNA Polymerase.
•Transcription of the operon is enhanced.
cAMP levels are influenced by glucose.

»The enzyme Adenylate cyclase catalyzes
the formation of cAMP and is
inhibited by glucose.
»When glucose is available in the cell,
Adenylate cyclase is inhibited and cAMP
levels are low.
» Under this condition CAP does not bind
upstream of the promoter and lac
operon is transcribed at a very low level.
When glucose is low, adenylate cyclase is not
inhibited, cAMP is higher and
CAP binds increasing the level of transcription.
»if lactose and glucose are present together ,
the lac operon will only transcribed
at a low level.
Trp Operon:
»The trp operon was discovered by Francis
Jacob and Jacques Monod in the early
1960s as part of their pioneering work on
generegulation.
» In the 1960s, Charles Yanofsky
demonstrated the existence of a mechanism
called
attenuation in Escherichia coli, where the
operon adjusts transcription in response to
tryptophan levels.
»It is a repressible operon that has two
independent genetic systems: one is
promoter-operator-trpE-trp-D-trp-C-trp-B-trp-A
system and another is the Promoter system.
In the former system, there is a leader region
termed as trp-L in between operator and trpE
gene.
Components of Trp Operon:
Repressor gene : The regulatory gene trp-R
encodes an repressor protein which is a
homodimer of two identical subunits, each with
107 amino acid residues.
•Regulatory region :
--Promoter : Tryptophan operon has two
promoter sites: one primary promoter P1 (40
nt)
located upstream of the trpE gene and another
secondary weak promoter P2 located at the
distal end of the trp-D gene between trp-D and
trpC genes.
--Operator : It is a 21 -bp sequence that is
located within the primary promoter. As the
operator and promoter completely overlap,
binding of repressor to the operator DNA
blocks
the RNA polymerase’s access to the promoter,
thereby inhibiting gene expression.
--Attenuator (leader region) : A second
regulatory sequence termed as trp leader
is
located between the operator and trpE gene.
This trp-L encodes a 162-nt long mRNA called
leader transcript that has four regulatory
regions (1 through 4) that can fold and form
secondary stem-loop or hair-pin structures by
complementary base pairing.
Structural genes :
-- trp E, trp D, trpC, trp-B, trp A
Regulation of Trp operon:

• In the absence of tryptophan.
• An inactive Repressor is made that cannot

bind to the operator (O), thus
allowing transcription to proceed.
• In the presence of tryptophan:
» Tryptophan binds to the repressor, causing
an allosteric transition to
occur. This complex binds to the operator
region, leading to repression
of the Operon.
Attenuation in trp operon:
The leader region of the mRNA contains :
1.Translation starts codon (AUG)
2. Two tryptophan codon UGG UGG
3. String of U residues at the end of the leader.
»The four regions marked 1,2,3 & 4 indicate
the location of four sequences
that have the potential to form stems by base
pairing.
»Region 2 is complementary to region 1 and 3.
» Region 3 is complementary to region 2 and
4.
--therefore several stem loop structures are

possible. Sequence of the trp-L mRNA
produced during attenuation
»The lack of tryptophan causes ribosomes to
stall at these codons, covering the region of
the mRNA required for formation of the
attenuation hairpin.
»This prevents attenuation and thus allows
transcription to continue.
Ara Operon:
»The ara operon is a genetic regulatory
system in Escherichia coli that controls the
metabolism of arabinose, a sugar.
»It allows E. coli to efficiently use arabinose as
a carbon source when it is available in the
environment.
»The three structural genes (araB, araA, araD)
encode for enzymes needed for the
metabolism of the sugar arabinose in bacterial
cells.
»araB, araA and araD encode for enzymes
kinase, isomerase and epimerase.
»Isomerase converts arabinose to ribulose.
»Kinase converts to ribulose 5 phosphate.
»The arabinose operon also contains the
Arabinose C gene which produces araC
protein.
The araC gene regulates the expression of the
structural genes and the araC
protein.Thus, the araC gene is autoregulated.
»The presence of both arabinose and araC
gene products activate the expression
of the BAD genes.
»The araC gene is adjacent to the ara operon.

--It has its own promoter, Pc
--It encodes a regulatory protein, AraC.
--AraC can bind to three different operator
sites,designated as araI, ara O 1 and ara O2
Negative regulation of ara operon:
»When Arabinose is absent, the AraC protein
acts as a negative regulator.
AraC acts as a dimer and causes the DNA
loop.
»Looping brings araI and ara O2 sites in
proximity to one another. »One
araC monomer binds to araI and a second
monomer binds to ara O2. » Binding
of araC prevents RNA Polymerase from
binding to the PBAD promoter.
»Transcription does not occur.
»Operon inhibited in the absence of
Arabinose:
Positive regulation of ara operon:

» When Arabinose is present, it binds to AraC
protein and changes AraC
conformation/ An arabinose-AraC dimer
complex binds preferentially to AraI and not to
the Ara O2 which causes the ‘opening’ of the
loop.
»This allows RNA Polymerase to bind to
PBAD
» If glucose levels are low, the cAMP-CAP
complex binds to the PC and activates the
transcription.
»Operon activated in the presence of
Arabinose
Summary of ara operon regulation:
★Levels of Arabinose is low and level of glucose

is high the operon is off,then th Cprotein bound to
Oand I,inhibiting transcription.
★Levels of Arabinose is low and level of glucose
is low ,the operon is on at figh frequency
Cprotein+; Arabinose bound to I and
cAM+CAP bound to I, enhancing transcription.

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OPERON SYSTEM

Positive control of lac operon:

cAMP levels are influenced by glucose.

Regulation of Trp operon:

• An inactive Repressor is made that cannot

--therefore several stem loop structures are

»The araC gene is adjacent to the ara operon.

Positive regulation of ara operon:

Summary of ara operon regulation:

★Levels of Arabinose is low and level of glucose

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