Gene Regulation
Gene Expression Is Controlled by Regulatory Proteins
Genes are very often controlled by extracellular signals; in the case of bacteria, this typically
means molecules present in the growth medium. These signals are communicated to genes
by regulatory proteins, which come in two types:
positive regulators, or activators, and negative regulators, or repressors. Typically, these
regulators are DNA-binding proteins that recognize specific sites at or near the genes they
control. An activator increases transcription of the regulated gene, and repressors decrease
or eliminate that transcription.
Many Promoters Are Regulated by Activators That Help RNA Polymerase Bind DNA and by
Repressors That Block That Binding At many promoters, in the absence of regulatory
proteins, RNA polymerase binds only weakly. This is because one or more of the promoter
elements
Gene Expression Is Controlled by Regulatory Proteins
An Activator and a Repressor Together Control the lac Genes. The three lac genes—lacZ, lacY,
and lacA—are arranged adjacently on the E. coli genome and are together called the lac
operon. The lac promoter, located at the 5’ end of lacZ, directs transcription of all three
genes as a single mRNA (called a polycistronic message because it includes more than one
gene); this mRNA is translated to give the three protein products.
The lacZ gene encodes the enzyme b-galactosidase, which cleaves the sugar lactose into
galactose and glucose, both of which are used by the cell as energy sources.
The lacY gene encodes the lactose permease, a protein that inserts into the cell membrane
and transports lactose into the cell.
The lacA gene encodes thiogalactoside transacetylase, which rids the cell of toxic
thiogalactosides that also get transported in by lacY.
These genes are expressed at high levels only when lactose is available, and glucose—the
preferred energy source—is not.
Two regulatory proteins are involved: one is an activator called CAP, and the other is a
repressor called the Lac repressor. The Lac repressor is encoded by the lacI gene, which is
located near the other lac genes, but transcribed from its own (constitutively expressed)
promoter.
The name CAP stands for catabolite activator protein, but this activator is also known as
CRP (for cAMP receptor protein) The gene encoding CAP is located elsewhere on the
bacterial chromosome, not linked to the lac genes. Both CAP and the Lac repressor are
DNA-binding proteins and each binds to a specific site on DNA at or near the lac
promoter.
Positive regulators (activators): Activator increases transcription of the regulated genes. In
lac operon, activator ( called CAP) acts as a glucose sensor. It activates the transcription of the
operon when glucose is absent/low.
Negative regulators (repressor): Repressor decreases or eliminates transcription of genes. In
lac operon, repressor acts as a lactose sensor. Lac repressor is encoded by the lacI gene. Lac
repressor prevents transcription of structural genes for lactose metabolism when lactose is
not available by tightly binding to the operator region.
In the absence of allolactose (A) the repressor protein (R) binds to the operator region (O)
and blocks the RNA polymerase from transcribing the structural genes.
Regulator Gene
PI Lac I Promoter Operator Lac Z Lac Y Lac A
CAP-binding site
Regulatory Structural gene
Regulator Gene
PI Lac I Promoter Operator Lac Z Lac Y Lac A
CAP-binding site
Lactose Transcription
Absent and Translation
Active Regulator
Repressor
Protein
Regulatory Structural gene
Regulator Gene
PI Lac I Promoter Operator Lac Z Lac Y Lac A
CAP-binding site
Lactose Transcription
Absent and Translation
Active Regulator
Repressor
Protein
RNA Polymerase
Regulatory Structural gene
Regulator Gene
PI Lac I Promoter Operator Lac Z Lac Y Lac A
CAP-binding site
Movement No
Lactose Transcription Blocked transcription
Absent and Translation
Active Regulator
Repressor
Protein
RNA Polymerase
Regulatory Structural gene
Regulator Gene
PI Lac I Promoter Operator Lac Z Lac Y Lac A
CAP-binding site
Lactose Transcription
Present and Translation
Glu Gal
Active Regulator
β-1,4
Repressor
Protein
Glu
Gal
β-1,6
Allolactose
When lactose is present
When lactose is present in a medium, some of it will be converted to allolactose. This
allolactose binds to the repressor protein. Binding of allolactose with repressor protein
changes the shape of repressor protein so it can no longer binds to the operator region. This
allows the RNA polymerase to bind to the promoter site, starting the initiation of
transcription of the structural genes lacZ, lacY, and lacA to produce mRNA.
Once the mRNA is produced, it is translated to
produce enzymes. The lacZ gene encodes for
the production of the enzyme β-galactosidase
(B), lacY gene for the production of the enzyme
permease (P) and the lacA gene is responsible
for the production of the enzyme
thiogalactoside transacetylase.
When lactose is present
RNA Polymerase
Regulatory Structural gene
Regulator Gene
PI Lac I Promoter Operator Lac Z Lac Y Lac A
CAP-binding site
Lactose Transcription Cant Bind to Operator
Transcription and Translation
Present and Translation
Glu Gal
β-1,4
Inactive β-galactosidase lactose permease thiogalactoside
Repressor transacetylase.
Glu
Glu Gal
Glu
Gal β-1,4
β-1,6 Gal
Allolactose
Catabolite repression (glucose effect)
Enzyme for lactose utilization can be induced by adding lactose in the growth medium but, it
would be wasteful to induce these enzymes if the cells are already growing on a carbon
source that they could use more efficiently e.g. glucose. This regulation is governed by a
phenomenon called catabolite repression, also known as glucose effect. Catabolite
repression ensures that the cells use the BEST carbon source first.
In the case of catabolite repressible enzymes, binding of RNA polymerase in the promoter
region of DNA occurs only if catabolite activator protein (CAP) also known as CRP (cAMP
Receptor Protein) has bound first. The gene encoding CAP is located elsewhere on the
bacterial chromosome, not linked to the lac genes. CAP is an allosteric protein which binds to
DNA only if it has first bound with cyclic AMP.
Cyclic AMP is synthesized from ATP by an enzyme called adenylate cyclase, and glucose
inhibits the synthesis of cyclic AMP and stimulates its transport out of the cell.
When glucose is transported into the cell, the cyclic AMP level in the cell is lowered. This
means only few CAP (which were bound with cyclic AMP) will be able to bind to DNA. This
will cause loose binding of RNA polymerase to the promoter region resulting low level of
transcription. When there is no any cyclic AMP left in the cell, the transcription will be
ceased.
Catabolite repression (glucose effect)
High glucose in the medium
When glucose levels are high, no cAMP is made, the CAP–cAMP complex does not form. CAP
cannot bind DNA without cAMP and RNA polymerase cannot bind to the promoter
efficiently. In this case, transcription occurs only at a low level.
adenylate cyclase
Inactive CAP
Glu cAMP
Low glucose in the medium
When the level of glucose in the environment is low or nil, abundant cAMP binds CAP to
form the CAP-cAMP complex, which binds DNA. When CAP–cAMP binds DNA, the efficiency
of RNA polymerase binding is increased at the lac operon promoter resulting in a higher level
of transcription of the structural genes.
adenylate cyclase
Active CAP
Glu cAMP
Regulation of Lac Operon
For the transcription of structural genes by the lac operon, two requirements must be met:
Glucose must be absent: The level of cyclic AMP must be high enough so that the CAP
protein binds to the CAP binding site. Bound CAP helps to attach RNA polymerase
efficiently to the lac operon promoter.
Lactose must be present: There must be an inducer (such as lactose) so that lactose
repressor does not block transcription by binding to the operator.
Each of the regulatory proteins (CAP and lac repressor) responds to one environmental
signal and communicates it to the lac genes. The combined effect of these two regulators
ensures that the genes are expressed at significant levels only when lactose is present and
glucose is absent.
Lactose Present
Glucose Absent
Active CAP
RNA Polymerase
PI Lac I Promoter Operator Lac Z Lac Y Lac A
CAP-binding site
Lactose Present
Glucose Absent
Active CAP
RNA Polymerase
PI Lac I Promoter Operator Lac Z Lac Y Lac A
CAP-binding site
Transcription
and Translation
Inactive Cant Bind to Operator
Repressor
Lactose Present
Glucose Absent
Active CAP
RNA Polymerase
PI Lac I Promoter Operator Lac Z Lac Y Lac A
CAP-binding site
Transcription and Translation
Transcription
and Translation
β-galactosidase lactose permease thiogalactoside
transacetylase.
Inactive Glu
Glu Gal
Repressor
β-1,4
Gal
Lactose Present
Glucose High
Inactive CAP
Cant Bind to CAP site
PI Lac I Promoter Operator Lac Z Lac Y Lac A
CAP-binding site
Low levels of Transcription and Translation
Transcription
and Translation
β-galactosidase lactose permease thiogalactoside
transacetylase.
Inactive Glu
Glu Gal
Repressor
β-1,4
Gal
Lactose Absent
Glucose High
Inactive CAP
Cant Bind to CAP site
PI Lac I Promoter Operator Lac Z Lac Y Lac A
CAP-binding site
Movement No Transcription
Transcription Blocked
and Translation
Active Regulator
Repressor
Protein
Lactose Absent
Glucose Absent
Active CAP
PI Lac I Promoter Operator Lac Z Lac Y Lac A
CAP-binding site
Movement No Transcription
Transcription Blocked
and Translation
Active Regulator
Repressor
Protein
References
• lehninger principles of biochemistry, Nelson David L., Albert L. Lehninger, David L. Nelson - 2005
• Molecular Biology of the Gene James D. Watson - 2004
• https://microbenotes.com/lac-operon/
• https://microbeonline.com/lac-operon-mechanism/
• https://microbeonline.com/lac-operon-mechanism/
