OPERON

A typical operon

Operon is a functioning unit of genomic DNA containing a cluster of genes under

the control of a single promoter. The genes are transcribed together into an
mRNA strand and either translated together in the cytoplasm, or undergo trans-
splicing to create monocistronic mRNAs that are translated separately, i.e. several
strands of mRNA that each encode a single gene product. The result of this is that
the genes contained in the operon are either expressed together or not at all.
Several genes must be co-transcribed to define an operon.

Types of operon:
There are different types of operon, they are, lactose operon, tryptophan operon
and galactose operon, etc.
The lac operon of the model bacterium Escherichia coli was the first operon to be
discovered and provides a typical example of operon function. It consists of three
adjacent structural genes, a promoter, a terminator, and an operator. The lac
operon is regulated by several factors including the availability of glucose and
lactose . It can be activated by allolactose . Lactose binds to the repressor protein
and prevents it from repressing gene transcription. This is an example of the de-
repressible.

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Lactose is the main sugar in milk, and they can be excellent meal for E.coli
bacteria. however, they will only gobble up lactose when other, better sugars
like glucose are not available.
Therefore, lac operon is an operon, or group of genes with a single promoter
(transcribed as a single mRNA). The genes in the operon encode proteins that
allow the bacteria to use lactose as an energy source.
E. coli bacteria can break down lactose, but it's not their favorite fuel. If glucose is
around, they would much rather use that. Glucose requires fewer steps and less
energy to break down than lactose. However, if lactose is the only sugar available,
the E. coli will go right ahead and use it as an energy source.

To use lactose, the bacteria must express the lac operon genes, which encode key
enzymes for lactose uptake and metabolism. To be as efficient as possible, E. coli
should express the lac operon only when two conditions are met:

Lactose is available, and

Glucose is not available

However this change affect lac operon transcription thereby providing two
regulatory protein, One, the lac repressor, acts as a lactose sensor.
The other, catabolite activator protein (CAP), acts as a glucose sensor.

These proteins bind to the DNA of the lac operon and regulate its transcription
based on lactose and glucose levels.

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STRUCTURE OF LACTOSE OPERON:
The lac operon contains three genes: lacZ, lacY, and lacA. These genes are
transcribed as a single mRNA, under control of one promoter.

Genes in the lac operon specify proteins that help the cell utilize lactose. lacZ
encodes an enzyme that splits lactose into monosaccharides (single-unit sugars)
that can be fed into glycolysis.

Similarly, lacY encodes a membrane-embedded transporter that helps bring

lactose into the cell.

The lacZ gene encodes an enzyme called -galactosidase, which is responsible for
splitting lactose (a disaccharide) into readily usable glucose and galactose
(monosaccharides).

The lacY gene encodes a membrane protein called lactose permease, which is a
trans-membrane "pump" that allows the cell to import lactose.

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The lacA gene encodes an enzyme known as a transacetylase that attaches a
particular chemical group to target molecules. It's not clear if this enzyme actually
plays any role in lactose breakdown. (Weird but true!)

In addition to the three genes, the lac operon also contains a number of
regulatory DNA sequences. These are regions of DNA to which particular
regulatory proteins can bind, controlling transcription of the operon.

The DNA of the lac operon contains CAP binding site, promoter (RNA polymerase
binding site), operator (which overlaps with promoter), lacZ gene, lacY gene, lacA
gene. The activator protein CAP bound to a molecule called cAMP, binds to the
CAP binding site and promotes RNA polymerase binding to the promoter.

TRYTOPHAN OPERON: The trp operon is an operona group of genes that is

used, or transcribed, togetherthat codes for the components for production of
tryptophan. The trp operon is present in many bacteria, but was first
characterized in Escherichia coli. The operon is regulated so that when tryptophan
is present in the environment, the genes for tryptophan synthesis are not
expressed. It was an important experimental system for learning about gene
regulation, and is commonly used to teach gene regulation. The trp operon in E.
coli was the first repressible operon to be discovered. While the lac operon can be
activated by a chemical (allolactose), the tryptophan (Trp) operon is inhibited by a
chemical (tryptophan).

In tryptophan operon, regulation of transcription occur at initiation and

termination. In E. coli the ve contiguous trp genes encode enzymes that

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synthesize the amino acid tryptophan. These genes are expressed efciently only
when tryptophan is limiting.

STRUCTURE OF TRYPTOPHAN OPERON

Trp operon contains five structural genes: trp E, trp D, trp C, trp B, and trp A,
which encode tryptophan synthetase. It also contains a repressive regulator gene
called trp R. trp R has a promoter where RNA polymerase binds and synthesizes
mRNA for a regulatory protein. The protein that is synthesized by trp R then binds
to the operator which then causes the transcription to be blocked. In the lac
operon, allolactose binds to the repressor protein, allowing gene transcription,
while in the trp operon, tryptophan binds to the repressor protein effectively
blocking gene transcription. In both situations, repression is that of RNA
polymerase transcribing the genes in the operon. Also unlike the lac operon, the
trp operon contains a leader peptide and an attenuator sequence which allows
for graded regulation.

This operon contains five structural genes Gene function trp R Codes for repressor
P, promoter O, Operator, site of attachment of RNA polymerase TrpE Codes for
Anthranllate TrpD Codes for Phosphoribosyl anthranllate transferase TrpC Codes
for Phosphoribosyl anthranllate isomerase TrpB Codes for Trp synthatase trpA
Codes for Trp synthatase

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PROKARYOTIC GENES REGULATION:

Gene Regulation is a process in which a cell determines which genes it will express
and when. Regulation of gene expression includes a wide range of mechanisms
that are used by the cell to increase or decrease the production of specific gene
products (protein or RNA). Sophisticated programs of gene expression are widely
observed in biology to trigger developmental pathways, respond to
environmental stimuli or adapt to new food sources. Regulation can occur at both
the initiation and termination of mRNA synthesis because bacteria obtain their
food from the medium that immediately surrounds them. Their regulation
mechanisms are designed to adapt quickly to the changing environment. If a gene
is not transcribed then the gene product and ultimately the phenotype will not be
expressed. In contrast, eukaryotes have much complex and larger genome and
cells of higher organisms are surrounded by constant internal milieu. The ability of
such cells to respond to hormones and to impulse in nervous system is thus
comparatively more important that the ability to respond rapidly in the presence
of certain nutrients. Diagram showing at which stages in the DNA-mRNA-protein
pathway expression can be controlled

Regulation of Transcription:

As in prokaryotes gene regulation occur at transcription level, so Transcription of

a gene by RNA polymerase can be regulated by at least five mechanisms;
Specificity factors alter the specificity of RNA polymerase for a given promoter or
set of promoters, making it more or less likely to bind to them (i.e., sigma factors
used in prokaryotic transcription). Repressors bind to the Operator, coding
sequences on the DNA strand that are close to or overlapping the promoter

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region, impeding RNA polymerase's progress along the strand, thus impeding the
expression of the gene. The image to the right demonstrates regulation by a
repressor in the lac operon. General transcription factors position RNA
polymerase at the start of a protein-coding sequence and then release the
polymerase to transcribe the mRNA. Activators enhance the interaction between
RNA polymerase and a particular promoter, encouraging the expression of the
gene. Activators do this by increasing the attraction of RNA polymerase for the
promoter, through interactions with subunits of the RNA polymerase or indirectly
by changing the structure of the DNA. Enhancers are sites on the DNA helix that
are bound by activators in order to loop the DNA bringing a specific promoter to
the initiation complex. Enhancers are much more common in eukaryotes than
prokaryotes, where only a few examples exist. Silencers are regions of DNA
sequences that, when bound by particular transcription factors, can silence
expression of the gene. Here, well explain two systems i.e, repression and
induction by which gene regulation is accomplished in prokaryotes.

PRINCIPLES OF TRANSCRIPTIONAL 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 specic sites at or near the genes they
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control. An activator increases transcription of the regulated gene; repressors
decrease or eliminate that transcription. First, RNA polymerase binds to the
promoter in a closed complex (in which the DNA strands remain together). The
polymerase-promoter complex then undergoes a transition to an open complex in
which the DNA at the start site of transcription is unwound and the polymerase is
positioned to initiate transcription.

Mechanism of Genes Regulation

The genes are controlled by a repressor, just as the lac genes are, but in this case
the ligand that controls the activity of that repressor (tryptophan) acts not as an
inducer but as a co-repressor. That is, when tryptophan is present, it binds the trp
repressor and induces a conformational change in that protein, enabling it to bind
the trp operator and prevent transcription. When the tryptophan concentration is
low, the trp repressor is free of its co-repressor and vacates its operator, allowing
the synthesis of trp mRNA to commence from the adjacent promoter.
Surprisingly, however, once polymerase has initiated a trp mRNA molecule it does
not always complete the full transcript. Indeed, most messages are terminated
prematurely before they include even the rst trp gene (trpE), unless a second
and novel device conforms that little tryptophan is available to the cell. Trp
operon; system turned on when tryptophan is absent, system turned off when
tryptophan is present.

conclusion: Trp operon and lac operon are two important biosynthetic systems in
E.coli that enable them to adapt quickly according to the changing environment.
Any imbalance or mutation at any stage or in any gene effect their activity and
ultimately their metabolic functions will be disturbed and they will not be able to

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cope up with the demands of the environment. Prokaryotes are unicellular or
colonial organisms. A number of genes are present in them but different genes in
the same cell are activated at different times. These alsso help the cell to spend a
lot of energy to activate different genes that are not required by the cell at
particular time. And in prokaryotes regulation is accomplished at transcription
level.

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 REFERENCE

Jacob, F., & Monod, J. On the regulation of gene activity. Cold Spring Harbor
Symposia on Quantitative Biology 26, 193211 (1962)

Lawrence, J. G. Shared strategies in gene organization among prokaryotes and

eukaryotes. Cell 11 William Klug, Cummings, and Spencer. "Concepts of
Genetics." 8th Ed. Pearson Education Inc, New Jersey: 2006. pg. 394-402

Yanofsky, C (1981). "Attenuation in the control of expression of bacterial operons".

Nature. 289: 751758. PMID 7007895. doi:10.1038/289751a0.

Lehninger, Albert L.; Nelson, David L.; Cox, Michael M. (2008). Principles of
Biochemistry (5th ed.). New York, NY: W.H. Freeman and Company. p. 1128.
ISBN 978-0-7167-7108-1.

SBertrand, K; Squires, C; Yanofsky, C (May 15, 1976). "Transcription termination in

vivo in the leader region of the tryptophan operon of Escherichia coli.".
Journal of Molecular Biology. 103 (2): 31937. PMID 781269.
doi:10.1016/0022-2836(76)90315-60, 407413 (2002

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