Prokaryotic Gene

Regulation

Enzymes are coded for

by genes
DNA is the code to make proteins
Enzymes are made of protein
In order for a cell to make an
enzyme, it must access the DNA for
that enzyme
Enzymes are very specific to their

DNA RNA Protein Trait

LE 17-3-2

TRANSCRIPTION

DNA

mRNA
Ribosome
TRANSLATION
Polypeptid
e

Prokaryotic
cell

Transcripti
on of DNA
to mRNA
starts at the
promoter,
ends at the
terminator

Some important
prerequisite facts
1. DNA is the code to make a protein
2. Some proteins are attracted to
specific sequences of DNA
3. Affinity for DNA sequences can
change with changes in protein
conformation
4. A special protein (RNA polymerase)
transcribes DNA RNA
5. Regulatory sequences of DNA
dont code for any specific protein,

Operons: The Basic

Concept

An operon is a collection of
prokaryotic genes transcribed together
on a single mRNA transcript 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

Basic Operon Regulation

Repressor Protein

RNA Polymerase

Repressor mRNA

Regulator
Gene

Promoter Operato
r

NO TRANSCRIPTION
Structural
Genes

Operon Regulations: Negative and positive

regulation

Repressible and Inducible Operons: Two

Types of Gene Regulation

An inducible operon is one that is

usually off; a molecule called an
inducer inactivates the repressor and
turns on transcription
The classic example of an inducible
operon is the lac operon
A repressible operon is one that is
usually on; binding of a repressor
shuts off transcription

 Inducible enzymes usually function in

catabolic pathways
Repressible enzymes usually function
in anabolic pathways
Regulation of the trp and lac operons
involves negative control of genes
because operons are switched off by
the active form of the repressor

 -galactosidase

10

 -galactosidase

H2O

galactose

lactose
g
- alactosidase

(aka lactase in humans)

glucose
11

-galactosidase Regulation
?
Why Regulategalactosidase
can
Levels at whichgalactosidase
regulated:
be
Genetic
Biochemical

12

Regulatory
gene

Lactose
Operon

Promoter
Operator

lacl

DNA

lacZ
No
RNA
made

3
mRNA
5

Protein

RNA
polymerase

Active
repressor

Lactose absent, repressor active, operon of

Lactose
Operon

LE 18-22b

lac operon
DNA

lacl

mRNA
5

lacZ

lacA

RNA
3 polymerase
mRNA 5

-Galactosidase Permease

Protein
Allolactose
(inducer)

lacY

Inactive
repressor

Lactose present, repressor inactive, operon on

Transacetylase

Positive Gene Regulation

Some operons are also subject to positive control
through a stimulatory activator protein, such as
catabolite activator protein (CAP)
When glucose (a preferred food source of E. coli )
is insufficient, the lac operon is activated by the
binding of CAP
When glucose levels increase, CAP detaches from
the lac operon, turning it off

Lactose
Operon
Promoter
DNA

lacl
CAP-binding site

Inactive
CAP

lacZ
Operator

RNA
polymerase
cant bind

Inactive lac
repressor

Lactose present, glucose present (cAMP level low): little lac

mRNA synthesized

Lactose
Operon

LE 18-23a

Promoter

DNA

lacZ

lacl
CAP-binding site

cAMP

Inactive
CAP

RNA
Operator
polymerase
Activecan bind
CAP and transcribe
Inactive lac
repressor

Lactose present, glucose absent (cAMP level high): abundant la

mRNA synthesized

LE 18-20

The
manufacture
of enzymes
responsible
for the
biosynthesis
of the amino
acid
tryptophan
is also
closely
regulated

Regulation of enzyme
activity
Precursor

Regulation of enzyme
production

Feedback
inhibition
Enzyme 1

Gene 1

Enzyme 2

Gene 2
Regulation
of gene
expression

Enzyme 3

Gene 3

Enzyme 4

Gene 4

Enzyme 5

Tryptophan

Gene 5

Prokaryotic Operon structure

ensures efficient regulation of
transcription
trp operon

Promoter

Promoter

DNA

mRNA

trpE

trpR

Regulatory
gene

trpD

trpC

trpB

trpA

Operator
Stop codon
RNA
Start codon
3 polymerase
mRNA 5

Protein

Genes of operon

E
Inactive
repressor

Polypeptides that make up

enzymes for tryptophan synthesis

Tryptophan absent, repressor inactive, operon on

The tryptophan biosynthesis operon is repressible by the

presence of its product, tryptophan

LE 18-21b_1

DNA

mRNA

Active
repressor

Protein

Tryptophan
(corepressor)
Tryptophan present, repressor active, operon of

LE 18-21b_2

DNA
No RNA made

mRNA

Active
repressor

Protein

Tryptophan
(corepressor)
Tryptophan present, repressor active, operon of

Tryptophan Operon
Tryptophan Present
Regulator
Gene

Promote
r

Operator Attenuator

RNA
Polymerase

trpR
mRNA
+
tryptophan
(corepress
TrpR protein or)
TrpR
(homodimer)
aporepressor
+ corepressor
(can bind to
operator)

Structural Genes

NO
TRANSCRIPTIO
N
Q: Why might the cell
want to produce an
aporepressor that is
only activated by the
operons end
product?

Tryptophan Operon
Tryptophan Absent
Regulator
Gene

Promote
r

Operator Attenuator

RNA
Polymerase

TRANSCRIPTIO
N

trpR
mRNA

TrpR protein
(homodimer)

Structural Genes

TrpR
aporepressor
(cannot bind
to operator)

Tryptophan Repressor
Protein
TrpR protein
subunits

Tryptophan
(corepressor)

DNA

Operons can be cut and pasted

together to make operon fusions
Tryptophan Operon
Repressor

Pro. Oper. Att.

Lactose Operon
lacI

TrpE, D, C, B, A

Promote Operator
r

Z
gene

T Pro. Oper. Z gene Y gene

Y gene

A gene

mRNA
mutant trpRcontaining
plasmid

-gal

A gene

If the repressor is knocked out,

what will happen in the presence
of Tryptophan?
Tryptophan Operon
Repressor

Pro. Oper. Att.

Lactose Operon
lacI

TrpE, D, C, B, A

Promote Operator
r

Z
gene

T Pro. Oper. Z gene Y gene

Y gene

A gene

mRNA
-gal

A gene

What if we add a plasmid which

contains the TrpR gene? With
tryptophan?
Without? Lactose Operon
Tryptophan Operon
Repressor

Pro. Oper. Att.

lacI

TrpE, D, C, B, A

Promote Operator
r

Z
gene

T Pro. Oper. Z gene Y gene

Y gene

A gene

mRNA
mutant trpRcontaining
plasmid

-gal

A gene