Introduction to Control of Gene Expression

• The controls that act on gene are much more complex in eukaryotes
than in prokaryotes.
• The presence of nuclear membrane in eukaryotes prevents the
simultaneous transcription and translation that occurs in prokaryotes.
• In prokaryotes, control of transcriptional initiation is the major point
of regulation.
• In eukaryotes the regulation of gene expression is controlled nearly
equivalently from many different points.

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 Control of gene expression in Prokaryotes

• In bacteria, genes are clustered into operons: gene clusters that encode the
proteins necessary to perform coordinated function, such as biosynthesis of a
given amino acid.
• RNA that is transcribed from prokaryotic operons is polycistronic a term
implying that multiple proteins are encoded in a single RNA molecule.
• In bacteria initiation is controlled by two DNA sequence elements –35 and –10
positions. These 2 sequence elements are termed promoter sequences, because
they promote recognition of transcriptional start sites by RNA polymerase.
• The consensus sequence for the -35 position is TTGACA, and for the –10
position, TATAAT. (The –10 position is also known as the Pribnow-box.) These
promoter sequences are recognized and contacted by RNA polymerase.
•
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Control of gene expression (continued)

• The activity of RNA polymerase at a given promoter is in turn

regulated by interaction with accessory proteins, which affect its
ability to recognize start sites. These regulatory proteins can act
both positively (activators) and negatively (repressors).

• The accessibility of promoter regions of prokaryotic DNA is in many

cases regulated by the interaction of proteins with sequences termed
operators. The operator region is adjacent to the promoter elements in
most operons and in most cases the sequences of the operator bind a
repressor protein.

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 Two major modes of transcriptional regulation in bacteria (E. coli)

• Both mechanisms involve repressor proteins. One mode of regulation is exerted

upon operons that produce gene products necessary for the utilization of energy;
these are catabolite-regulated operons. The other mode regulates operons that
produce gene products necessary for the synthesis of small biomolecules such as
amino acids. Expression from the latter class of operons is attenuated by
sequences within the transcribed RNA.
• A classic example of a catabolite-regulated operon is the lac operon,
responsible for obtaining energy from β-galactosides such as lactose.
• A classic example of an attenuated operon is the trp operon, responsible for the
biosynthesis of tryptophan.
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 LAC OPERON
• The lac operon (see diagram below) consists of one regulatory gene (the i gene) and three structural genes
(z, y, and a). The i gene codes for the repressor of the lac operon. The z gene codes for β-galactosidase
(β-gal), which is primarily responsible for the hydrolysis of the disaccharide, lactose into its monomeric
units, galactose and glucose. The y gene codes for permease, which increases permeability of the cell to B
Galatoside such as lactose . The a gene encodes a transacetylase, it function is detoxification.
• During normal growth on a glucose-based medium, the lac repressor is bound to the operator region of
the lac operon, preventing transcription. However, in the presence of an inducer of the lac operon, the
repressor protein binds the inducer and is rendered incapable of interacting with the operator region of the
operon. RNA polymerase is thus, able to bind at the promoter region, and transcription of the operon
ensues.
• The lac operon is repressed, even in the presence of lactose, if glucose is also present. This repression
is maintained until the glucose supply is exhausted. The repression of the lac operon under these
conditions is termed catabolite repression and is a result of the low levels of cAMP that result from an
adequate glucose supply.
•

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Lac operon (continued)
• The repression of the lac operon is relieved in the presence of glucose if excess cAMP
is added. As the level of glucose in the medium falls, the level of cAMP increases.
Simultaneously there is an increase in inducer binding to the lac repressor. The net
result is an increase in transcription from the operon.

• The ability of cAMP to activate expression from the lac operon results from an
interaction of cAMP with a protein termed CRP (for cAMP receptor protein). The
protein is also called CAP (for catabolite activator protein). The cAMP-CRP
complex binds to a region of the lac operon just upstream of the region bound by RNA
polymerase and that somewhat overlaps that of the repressor binding site of the
operator region.
• The binding of the cAMP-CRP complex to the lac operon stimulates RNA polymerase
activity 20–50 fold.
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Regulation of the lac operon in E. coli.

• The repressor of the operon is synthesized from the i gene.

• The repressor protein binds to the operator region of the operon and prevents RNA
polymerase from transcribing the operon.
• In the presence of an inducer (such as the natural inducer, allolactose) the
repressor is inactivated by interaction with the inducer. This allows RNA
polymerase access to the operon and transcription proceeds. The resultant mRNA
encodes the β-galactosidase, permease and transacetylase activities necessary for
utilization of β-galactosides (such as lactose) as an energy source. The lac operon
is additionally regulated through binding of the cAMP receptor protein, CRP (also
termed the catabolite activator protein, CAP) to sequences near the promoter
domain of the operon. The result is a 50 fold enhancement of polymerase activity
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