Lac Operon

By Azrin Nahian
The Lac Operon

The lactose operon (lac operon) is an operon required for the transport and
metabolism of lactose in E.coli and many other enteric bacteria. Although glucose is
the preferred carbon source for most bacteria, the lac operon allows for the effective
digestion of lactose when glucose is not available through the activity of beta-
galactosidase.

The lac operon is a negatively controlled inducible operon, where the inducer
molecule is allolactose. In negative repressible operons, transcription of the operon
normally takes place.

Repressor proteins are produced by a regulator gene, but they are unable to bind to
the operator in their normal conformation.
Structure of Lac Operon
Structure of Lac 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. 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 promoter is the binding site for RNA polymerase, the enzyme that performs
transcription.
• The operator is a negative regulatory site bound by the Iac repressor protein. The
operator overlaps with the promoter,
and when the Iac repressor is bound, RNA polymerase cannot bind to the promoter and
start transcription.
• The CAP binding site is a positive regulatory site that is bound by catabolite activator
protein (CAP). When CAP is bound to this site, it promotes transcription by helping RNA
polymerase bind to the promoter. Let's take a closer look at the lac repressor and CAP and
their roles in regulation of the lac operon.
So, when does the lac operon
really turn on?

The lac operon will be expressed at high levels if two conditions are met:
• Glucose must be unavailable: When glucose is unavailable, cAMP binds to CAP,
making CAP able to bind DNA. Bound CAP helps RNA polymerase attach to the lac
operon promoter.
• Lactose must be available: If lactose is available, the lac repressor will be released
from the operator (by binding of allolactose. This allows RNA polymerase to move
forward on the DNA and transcribe the operon.

These two events in combination - the binding of the activator and the release of the
repressor - allow RNA polymerase to bind strongly to the promoter and give it a clear
path for transcription. They lead to strong transcription of the lac operon and production
of enzymes needed for lactose utilization.
 Putting it all together
Now that we've seen all the moving parts of the lac operon, let's put what we've learned together to see how the operon reacts to a
variety of different conditions (presence or absence of glucose and lactose).

• Glucose present, lactose absent: No transcription Of the lac
operon occurs. That's because the lac repressor remains bound
to the operator and prevents transcription by RNA polymerase.
Also, cAMP levels are low because glucose levels are high. so
CAP is inactive and cannot bind DNA.
•Glucose absent, lactose absent: Glucose absent, lactose absent:
No transcription of the lac operon occurs. cAMP levels are high
because glucose levels are low, so CAP is active and will be
bound to the DNA. However, the lac repressor will also be bound
to the operator (due to the absence of allolactose), acting as a
roadblock to RNA polymerase and preventing transcription.
 Putting it all together
• Glucose present, lactose present: Low- level transcription of
the Iac operon occurs. The Iac repressor is released from the
operator because the inducer (allolactose) is present. cAMP
levels, however, are low because glucose is present. Thus,
CAP remains inactive and cannot bind to DNA, so
transcription only occurs at a low, leaky level.
• Glucose absent, lactose present: Strong transcription of the
lac operon occurs. The lac repressor is released from the
operator because the inducer (allolactose) is present. cAMP
levels are high because glucose is absent, so CAP is active and
bound to the DNA. CAP helps RNA polymerase bind to the
promoter, permitting high levels of transcription.
Summary of lac operon Responses

Repressor Level of
Glucose Lactose CAP Binds
Binds Transcription

_ _ No
+ + Transcription

+ + _ _ Low-level
Transcription

_ _ No
+ +
Transcription

_ _ Strong
+ +
Transcription