Regulation of Gene Expression

in Prokaryotes
(KCSP 17)

Genetics 244
Introductory Molecular Biology

Departement Genetika

Genetics Department
 Regulation of Gene Expression in Prokaryotes

Efficient expression of genetic information is

dependant on control mechanisms that promote or
Genetics 244 – Gene Regulation in Prokaryotes

suppress gene activities.

• Studies were performed on bacteria and yeast: easy to

culture and mutate, fluctuating protein levels
• Adaptation hypothesis: lactose in yeast growth medium
induces expression of enzymes specific for lactose
metabolism
• Constitutive, inducible, repressible systems,
negative/positive control
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 Genetics 244 – Gene Regulation in Prokaryotes

the big picture…

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 Inducible system:
Lactose metabolism in E. coli

• Lactose = galactose +
Genetics 244 – Gene Regulation in Prokaryotes

glucose

• Enzymes responsible for

lac metabolism:
5-10 copies vs. 1000s in
presence of lactose inducer

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 The lac operon
• Gene complex (operon) consists of 3 structural genes
and 2 regulating “genes” (cis & trans action)
Genetics 244 – Gene Regulation in Prokaryotes

• Structural genes (related functions):

• lac Z – -galactosidase
• lac Y – -galactoside permease
• lac A – transacetylase

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 The lac operon
• Polycistronic mRNA: order Z-Y-A
Genetics 244 – Gene Regulation in Prokaryotes

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 How does lactose regulate
transcription of the 3 structural genes?
• Gene activity repressed in absence of lactose –
lactose induces
Genetics 244 – Gene Regulation in Prokaryotes

• Not through interaction with enzymes – IPTG

(gratuitous inducer)
• Constitutive mutations – lac I - and lac Oc
isopropylthiogalactoside
Operon model (negative control)

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 Genetics 244 – Gene Regulation in Prokaryotes

Functioning of the lac operon

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 Mutations in the lac operon
Genetics 244 – Gene Regulation in Prokaryotes

Example of negative
control
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 Genetics 244 – Gene Regulation in Prokaryotes
s
Lac I mutation

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 Genetic proof for the Operon Model
• Hypothesis:
Genetics 244 – Gene Regulation in Prokaryotes

• lacI produces a diffusible product

• lacO has no gene product
• lacO must be adjacent to the structural genes in order to
regulate them

• Partially diploid bacteria – F’-plasmids (Table 16.1)

• Repressor isolated and proven to be a protein

(4 subunits, 38kDa each)
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 Lac operon gene activity
-Galactosidase activity
Genotype Lactose present Lactose absent
Genetics 244 – Gene Regulation in Prokaryotes

I+O+Z+ + -
I+O+Z- - -
I-O+Z+ + +
I+OCZ+ + +
I-O+Z+ / F’ I+ + -
I+OCZ+ / F’ O+ + +
I+O+Z+ / F’ I- + -
I+O+Z+ / F’ OC + -
ISO+Z+ - -
ISO+Z+ / F’ I+
- - 12
 Positive control:
Catabolite Activating Protein
What happens if glucose is present in abundance in the
environment?
• catabolite-activating protein (CAP) involved in the repression of
Genetics 244 – Gene Regulation in Prokaryotes

expression of the lac operon

• catabolite repression
• CAP binding on 5’ side of the promotor
• effective promotor-binding only in presence of cAMP
adenyl cyclase
ATP cAMP
• Glucose inhibits adenyl cyclase
• Thus: In the presence of glucose – poor CAP binding, poor Rp
binding and resultantly poor transcription/translation
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• Example of positive control – facilitates Rp binding
 Genetics 244 – Gene Regulation in Prokaryotes

CAP

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 Tryptophan metabolism:
A repressible system
• Presence of tryptophan in environment results in repression of
tryptophan synthesis
Genetics 244 – Gene Regulation in Prokaryotes

• 5 enzymes, encoded by 5 consecutive genes (trp E, trp D, trp

C, trp B, trp A), are responsible for tryptophan synthesis

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 Functioning of the trp operon
• Hypothesis – inactive repressor cannot interact with operator
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 Functioning of the trp operon
• In the presence of tryptophan – allosteric transition; now
possible to bind to operator and prevent transcription
• Example of negative control
Genetics 244 – Gene Regulation in Prokaryotes

• tryptophan is a co-repressor
• attenuation

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 Arabinose regulating protein:
Positive and negative control
• Arabinose metabolism – ara B, ara A and ara D structural genes
• ara C gene – regulating protein (dimer)
Genetics 244 – Gene Regulation in Prokaryotes

• Operator region
• Inducer site with CAP binding site

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 Functioning of the ara operon
• Arabinose present
• protein only binds to the I-region – serves as a inducer;
Genetics 244 – Gene Regulation in Prokaryotes

positive control
• cAMP present – CAP binds

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 Functioning of the ara operon
• Arabinose absent
• protein binds to I-region and O2-site – serves as a repressor;
Genetics 244 – Gene Regulation in Prokaryotes

negative control

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