LECTURE 7

REGULATION OF GENE EXPRESSION

(PROTEIN SYNTHESIS)
 Prokaryotic Regulation
• Enzymes and other proteins are required by
cells for particular metabolic reactions and in
specific concentrations.
• Bacterial cells need to control protein
production so that there is no wastage of
energy, protein or enzymes that are not
required by the cell.
• Bacterial cells achieve this in three different
ways;
(i) Rapid degradation of mRNA soon after
translation.
(ii) By allosteric interaction especially with
enzymes.
(iii) By transcribing all enzymes involved in
biosynthesis on a single molecule of mRNA.
Such a group of functionally related genes
occuring on a single molecule of mRNA is
called an operon.
• For example, the biosynthesis of an amino
acid histidine requires enzymes that work in
sequence for a specific reaction pathway.
• The genes for such enzymes are all transcribed
on a single mRNA molecule.
• The expression of such an operon is tightly
controlled in that the synthesis of histidine
enzymes are only synthesized when there is
an insufficient level of histidine available in
the cell.
• An operon model explains gene regulation in
prokaryotes.
• An operon mechanism typically includes the
following elements;
1. Promoter: A short sequence of DNA where RNA
polymerase first attaches to begin transcription
of the grouped genes.
• A promoter signals where transcription is to
begin.
2. Operator: A short portion of DNA sequence
where an active repressor binds.
• When an active repressor binds to the
operator site, RNA polymerase can not attach
to the promotor site and transcription can not
occur.
3. Structural genes: One to several genes coding
for the primary structure of enzymes in a
metabolic pathway transcribed as a unit.
• Regulator gene: Normally located outside the
operon and controlled by its own promoter,
codes for a repressor protein that controls
whether the operon is active or not.
 Types of Regulations
• Up-regulation is the process in which an
internal or external signal results in increased
expression of one or more genes resulting in
increased protein synthesis.
• Down-regulation is the process resulting in
decreased gene expression and a
corresponding decrease in protein synthesis.
Inducible and Repressible systems of
gene expression
• An inducible system is normally switched off
and is only switched on in the presence of an
inducer molecule.
• The induce is said to induce expression.
• A repressible system is usually switched on
except in the presence of a co-repressor
molecule that suppresses gene expression.
• The co-repressor is said to repress expression.
 The Lac Operon
• The lac operon is a bio-synthesis system that
is required for the efficient transport and
metabolism of lactose (milk sugar) in
Escherichia Coli and other related bacteria.
• In the absence of glucose, the E. coli bacteria
cells can utilise lactose as a source of energy
however it has to be processed via the lac
operon.
• The lac operon is a set of three structural genes
that are linked in sequence on one strand of
DNA.
• The three genes are called Lac Z, Lac Y and Lac
A.
• They code for enzyme proteins called β-
galactosidase, β-galactoside permease and β-
galactoside transacetylase respectively.
• These are the structural genes of the operon
because they code for specific proteins.
• They are activated by a single promoter and
produce a single mRNA molecule.
• The same sequence of nucleotide bases which
constitute an operon also carry non structural
genes that do not code for protein but aid the
operon in regulating the metabolism of lactose.
• These extra genes on the operon are collectively
called regulator sites.
• Transcription of the regulatory genes produce
an mRNA which is used to synthesis a repressor
protein.
• Therefore, a lac operon is a sequence of
nucleotide bases divided into three structural
genes, the operator and promotor site for
metabolism of lactose.
• Specific control of the lac genes depends on
the presence or absence of lactose in a cell.
• When lactose is absent in the in the bacteria
growth, the lac gene remain dormant because
the need to produce enzymes required to
process the lactose does not arise.
• When lactose is present enzyme β-galactoside
permease insert in the membrane and allows
lactose to flood into the cell at a faster rate.
• Once the lactose is inside the cell enzyme β-
galactosidase breaks down lactose into simpler
sugars glucose and galactose.
• These simple sugars are the used as source of
energy by the cells.
• Enzyme β-galactoside transacetylase facilitates
the transfer of acetyl group from acetyl-CoA to
β-galactoside .
• If lactose is missing from the growth medium of
a bacterium, DNA outside the lac operon
transcribes mRNA to synthesise a protein called
Lac repressor protein.
• This repressor protein travels to the lac operon
on the DNA and binds to a lac operator site.
• This action by the repressor protein interferes
with binding of RNA polymerase to the
promoter site.
• The repressor blocks the locomotive progression
of RNA polymerase along the lac operon on the
DNA molecule.
• Therefore, RNA polymerase is unable to
transcribe the lac structural genes and the
corresponding protein of the operon are not
synthesised.
• When the lactose is added to the bacteria cell
medium, the substrate (lactose) enters the cell
passively and some of it (allolactose) attaches
to the allosteric site of the lac repressor
protein.
• The lac repressor protein loses its shape and
can no longer fit into the operator site on the
DNA.
• The lac repressor protein finally rolls out
• Since there is no more interference along the
lac operon, RNA polymerase can attaches to
the promoter site and transcribe the lac
operon genes on to the mRNA as it moves
along the DNA from 3’ to 5’.
• As a result, a polycistronic mRNA is
manufactured which encodes for the three
three lac structural genes.
• As the lactose substrate runs out in the
medium, its absence induces a repressor
protein to go back to the original shape and
bind to the DNA operator site once again.
END OF LECTURE!
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