Operon concept

Discovery of the Operon

In 1961, Jacob and Monod studied the metabolism of lactose

by E. coli
Three enzyme activities / three genes were induced together

by galactosides
Based on their observation they propossed operon concept

Nobel prize in the year 1965

Introduction
An operon is a collection of prokaryotic genes

transcribed together on a single mRNA transcript

(polycistronic) to serve a single purpose
Composed of

An operator, an on-off switch

A promoter

Genes for metabolic enzymes

Can be switched off by a repressor protein

A corepressor is a small molecule that binds to

a repressor to switch an operon off

Regulatory proteins control operon
transcription:
Operons are either REPRESSIBLE or
INDUCIBLE

REPRESSIBLE: INDUCIBLE:

Involvesbindinga Involvesbindingan
corepressor inducer
(metabolite/end (metabolite/starting
product)to substrate)to
repress/turnOFF induce/turnON
operontranscription operontranscription
 3 combinations found in nature:

REPRESSIBLE INDUCIBLE

Thisoperonusesa Thisoperonusesa
repressorthatbindsaco repressorthatbindsan
NEGATIVE repressortorepress inducertoinduce
transcription. transcription.

Thisoperonusesan
activatorthatbindsan
POSITIVE Notfound.
inducertoinduce
transcriptionn.
 Lac operon

The lac operon was the first operon discovered

It contains 3 genes coding for E. coli proteins that

permit the bacteria to use the sugar lactose

LacZ encodes Beta-galactosidase
breaks up lactose into glucose and galactose (galactose also
converted to glucose for metabolism)
Isomerizes lactose into allolactose inducer (presence of lactose
means presence of allolactose)
LacY encodes permease
For lactose transport across cell membrane
LacA encodes transacetylase
Poorly understood function
The Lac operon has 2 control
circuits:
NEGATIVE INDUCIBLE
Uses a repressor that binds an inducer
(allolactose) to induce operon
transcription.
Requires presence of lactose

POSITIVE INDUCIBLE
Uses an activator (CAP) that binds an
inducer (cyclic AMP) to induce operon
transcription.
Requires absence of (preferred) glucose
LAC OPERON NEGATIVE INDUCIBLE CONTROL CIRCUIT:
In the absence of lactose, an active repressor
protein binds to the operator and blocks
transcription by RNA Polymerase:
When lactose is present in the cell, allolactose, an
isomer of lactose, binds to the repressor.
This inactivates the repressor, because it can no
longer bind the operator.
Now RNA Polymerase can transcribe the Lac operon:
RNA polymerase have low

affinity towards lac operon

So even the operon is on

transcription will occur poorly

There comes the role of positive

inducible control
LAC OPERON POSITIVE INDUCIBLE
CONTROL CIRCUIT
Attachment of RNA polymerase to the promoter requires

the presence of a Catabolite gene Activatior Protein

(CAP) that is bound to the cAMP
Presence of glucose lowers the cAMP by inactivating

adenylyl cyclase responsible for the synthesis of cAMP

Due to lesser cAMP formation of CAP-cAMP is low

So RNA polymerase attachment and transcription will be

negligible in the presence of glucose

As CAP-cAMP is essential for transcription of lac operon

it is considered as positive regulator.

Structure of IPTG

Figure 10-7
 Trp Operon

TrpE gene product

TrpD gene product

TrpC gene product

TrpB gene product

TrpA gene product

The Trp operon has 2 control
mechanisms
NEGATIVE REPRESSIBLE OPERON
Uses a repressor that binds a co-repressor
(end product Trp) to repress operon
transcription by 70-fold
Requires presence of Trp

ATTENUATION
Involves premature transcription termination
Requires high Trp levels
TRP OPERON NEGATIVE REPRESSIBLE CONTROL CIRCUIT: By itself, the
operon is on. RNA polymerase can bind to the promotor and moves freely
through the operator to transcribe the genes:
When co-repressor (end-product) Trp is present,
it binds to the repressor. This activates the
repressor, causing it to bind the operator to block
Trp operon transcription: