Department of Biological Sciences

College of Arts and Sciences

VISAYAS STATE UNIVERSITY

Visca, Baybay City, Leyte, 6521-A, Philippines

WRITTEN REPORT ON THE LACTOSE (lac) OPERON

I. Introduction

 Escherichia coli is able to use other sugars as energy sources when glucose
concentrations are low. One such sugar source is lactose, which is found in
milk.
 The lac operon (lac for lactose) encodes three genes necessary to acquire and
process the lactose from the local environment.
 lacZ (or simply Z gene) – encodes β-galactosidase, which breaks lactose
down to glucose and galactose and can also convert lactose into
allolactose (an isomer of lactose).
 lacY (or Y gene)– encodes for the enzyme permease that is used to
actively transport lactose into the cell.
 lacA (or A gene) – used to produce the third enzyme, thiogalactoside
transacetylase, whose function is not yet known but is supposed to play
a role in the removal of toxic by-products of lactose digestion by the cell.
 The entire transcription unit is under the command of one main operator (lacO)
and promoter (lacP). The regulatory gene, (lacI), located outside the operon
and has its own promoter (PI), codes for an allosteric repressor protein that can
switch off the lac operon by binding to the operator.
 The repressor consists of four identical polypeptides and has two types of
binding sites; one type of site binds to allolactose and the other binds to DNA.
In the absence of lactose (and, therefore, allolactose), the repressor binds to
the lac operator site lacO.

II. Regulation of the lac operon

A. Negative Gene Regulation and Induction
 Unlike the trp repressor, the lac repressor is active by itself, binding to the
operator and switching the lac operon off. In this case, a specific small
molecule, called an inducer, inactivates the repressor. For the lac operon, the
inducer is allolactose, formed in small amounts from lactose that enters the
cell.
In the absence of lactose

 The lac repressor is in its active shape. It binds to the operator and blocks the
RNA polymerase, preventing transcription and the genes of the lac operon are
silenced. Only few molecules of each enzymes are produced.
In the presence of lactose

 Allolactose binds to the lac repressor and alters its shape, nullifying the
repressor’s ability to attach to the operator and the binding of RNA polymerase
is no longer blocked. The transcription of lacZ, lacY, and lacA takes place and
the lac proteins (enzymes) are produced.
 Without the repressor bound, the lac operon is transcribed into mRNA for the
lactose-utilizing enzymes.
 In the context of gene regulation, the enzymes of the lactose pathway are
referred to as inducible enzymes because their synthesis is induced by a
chemical signal (allolactose, in this case). The regulation of lac operon also
involves the negative control of genes because the operon is switched off by
the binding of the active form of the repressor protein. Therefore, the lac
operon is a negative inducible operon.

B. Positive Gene Regulation and Glucose Effect

Glucose repression (glucose effect) – a mechanism for the preferential use of
glucose in the presence of other less efficient energy sources, shutting down genes
that participate in the metabolism of other sugars such as lactose

 When glucose and lactose are both present in its environment, E. coli
preferentially uses glucose because glucose enters glycolysis without further
modification and therefore requires less energy to metabolize than do other
sugars.
 Only when lactose is present and glucose is in short supply does E. coli use
lactose as an energy source, and only then does it synthesize appreciable
quantities of the enzymes for lactose breakdown.
When glucose is scarce

 The mechanism depends on the interaction of an allosteric regulatory protein

with a small organic molecule, the cyclic AMP (cAMP), which accumulates
when glucose is scarce. The regulatory protein, called catabolite activator
protein (CAP), is an activator. (Note: Glucose inhibits the activity of adenyl
cyclase, affecting the level of cAMP in the cell)
 When cAMP binds to this regulatory protein, CAP assumes its active shape and
can attach to a specific site at the upstream end of the lac promoter. This
attachment increases the affinity of RNA polymerase for the promoter, which
is actually rather low even when no repressor is bound to the operator. The
 cAMP-CAP complex must be present at its binding site in the lac promoter in
order for the operon to be induced normally
 By facilitating the binding of RNA polymerase to the promoter and thereby
increasing the rate of transcription, the attachment of CAP to the promoter
directly stimulates gene expression.
When glucose is high

 The cAMP concentration falls, and without cAMP, CAP detaches from the
operon. Only the cAMP-CAP complex binds to the lac promoter; in the absence
of cAMP, CAP does not bind. Thus, cAMP acts as the effector molecule,
determining the effect of CAP on lac operon transcription. It could also be that
only little cAMP-CAP complex is available to bind to the DNA.
 Because CAP is inactive, RNA polymerase binds less efficiently to the promoter,
and transcription of the lac operon proceeds only at a low level, even when
lactose is present.

C. Conclusion
Therefore, the lac operon is under dual control: negative control by the lac
repressor and positive control by CAP. The state of the lac repressor (with or without
bound allolactose) determines whether or not transcription of the lac operon’s genes
occurs at all; the state of CAP (with or without bound cAMP) controls the rate of
transcription if the operon is repressor-free. It is as though the operon has both an
on-off switch and a volume control.

For the lac operon to be activated, two conditions must be met.

1. The level of glucose must be very low or non-existent.

2. Second, lactose must be present.

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