The lac operon:

Regulation of genes for lactose utilization. lac repressor, catabolite activator protein, and
cAMP.

Key points:

The lac operon of E. coli contains genes involved in lactose metabolism. It's expressed
only when lactose is present and glucose is absent.

Two regulators turn the operon "on" and "off" in response to lactose and glucose
levels: the lac repressor and catabolite activator protein (CAP).

The lac repressor acts as a lactose sensor. It normally blocks transcription of the
operon, but stops acting as a repressor when lactose is present. The lac repressor

senses lactose indirectly, through its isomer allolactose.

Catabolite activator protein (CAP) acts as a glucose sensor. It activates transcription
of the operon, but only when glucose levels are low. CAP senses glucose indirectly,
through the "hunger signal" molecule cAMP.

Introduction
Lactose: it's what's for dinner! While that may not sound delicious to us (lactose is the
main sugar in milk, and you probably don't want to eat it plain), lactose can be an
excellent meal for E. coli bacteria. However, they'll only gobble up lactose when other,
better sugars like glucose are unavailable.
With that for context, what exactly is the lac operon? The lac operon is anoperon, 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.

What makes the lac operon turn on?

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

How are levels of lactose and glucose detected, and how how do changes in levels
affect lac operon transcription? Two regulatory proteins are involved:

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. Let's take a look at how this works.

Structure of the 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.
[More details]

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.

Structure of the lac operon. The DNA of the lac operon contains (in order from left to
right): CAP binding site, promoter (RNA polymerase binding site), operator (which

overlaps with promoter), lacZ gene, lacY gene, and lacA gene. The activator protein
CAP, when bound to a molecule called cAMP (discussed later), binds to the CAP
binding site and promotes RNA polymerase binding to the promoter. The lac repressor
protein binds to the operator and blocks RNA polymerase from binding to the
promoter and transcribing the operon.
Image modified from "Prokaryotic gene regulation: Figure 3," by OpenStax College, Biology (CC BY 4.0).

The promoter is the binding site for RNA polymerase, the enzyme that performs
transcription.

The operator is a negative regulatory site bound by the lac repressor protein. The
operator overlaps with the promoter, and when the lac 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.

The lac repressor

The lac repressor is a protein that represses (inhibits) transcription of the lac operon. It
does this by binding to the operator, which partially overlaps with the promoter. When
bound, the lac repressor gets in RNA polymerase's way and keeps it from transcribing
the operon.
[Where does the lac repressor come from?]

When lactose is not available, the lac repressor binds tightly to the operator,
preventing transcription by RNA polymerase. However, when lactose is present,
the lac repressor loses its ability to bind DNA. It floats off the operator, clearing the
way for RNA polymerase to transcribe the operon.

Upper panel: No lactose. When lactose is absent, the lac repressor binds tightly to the
operator. It gets in RNA polymerase's way, preventing transcription.
Lower panel: With lactose. Allolactose (rearranged lactose) binds to the lac repressor
and makes it let go of the operator. RNA polymerase can now transcribe the operon.
Image modified from "Prokaryotic gene regulation: Figure 3," by OpenStax College, Biology (CC BY 4.0).

This change in the lac repressor is caused by the small molecule allolactose, an isomer
(rearranged version) of lactose. When lactose is available, some molecules will be
converted to allolactose inside the cell. Allolactose binds to the lac repressor and
makes it change shape so it can no longer bind DNA.
Allolactose is an example of an inducer, a small molecule that triggers expression of a
gene or operon. The lac operon is considered an inducible operon because it is

usually turned off (repressed), but can be turned on in the presence of the inducer
allolactose.

Catabolite activator protein (CAP)

When lactose is present, the lac repressor loses its DNA-binding ability. This clears
the way for RNA polymerase to bind to the promoter and transcribe the lac operon.
That sounds like the end of the story, right?
Well...not quite. As it turns out, RNA polymerase alone does not bind very well to
the lac operon promoter. It might make a few transcripts, but it won't do much more
unless it gets extra help from catabolite activator protein (CAP). CAP binds to a
region of DNA just before the lac operon promoter and helps RNA polymerase attach
to the promoter, driving high levels of transcription.
[Where does CAP come from?]

Upper panel: Low glucose. When glucose levels are low, cAMP is produced. The
cAMP attaches to CAP, allowing it to bind DNA. CAP helps RNA polymerase bind to
the promoter, resulting in high levels of transcription.
Lower panel: High glucose. When glucose levels are high, no cAMP is made. CAP
cannot bind DNA without cAMP, so transcription occurs only at a low level.
Image modified from "Prokaryotic gene regulation: Figure 3," by OpenStax College, Biology (CC BY 4.0).

CAP isn't always active (able to bind DNA). Instead, it's regulated by a small molecule
called cyclic AMP (cAMP). cAMP is a "hunger signal" made by E. coli when glucose
levels are low. cAMP binds to CAP, changing its shape and making it able to bind
DNA and promote transcription. Without cAMP, CAP cannot bind DNA and is
inactive.
[How is cAMP made, and how does it report glucose levels?]

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CAP is only active when glucose levels are low (cAMP levels are high). Thus,
the lac operon can only be transcribed at high levels when glucose is absent. This
strategy ensures that bacteria only turn on the lac operon and start using lactose after
they have used up all of the preferred energy source (glucose).

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 weve seen all the moving parts of the lac operon, lets put what weve
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 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.
Image modified from "Prokaryotic gene regulation: Figure 3," by OpenStax College, Biology (CC BY 4.0).

Glucose present, lactose present: Low-level transcription of the lac operon occurs.
The lac 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 present, lactose present: Low-level transcription of the lac operon occurs.
The lac 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.
Image modified from "Prokaryotic gene regulation: Figure 3," by OpenStax College, Biology (CC BY 4.0).

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.

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.
Image modified from "Prokaryotic gene regulation: Figure 3," by OpenStax College, Biology (CC BY 4.0).

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.

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.
Image modified from "Prokaryotic gene regulation: Figure 3," by OpenStax College, Biology (CC BY 4.0).

Summary of lac operon responses

Glucose

Lactose CAP binds Repressor binds

Level of transcription

No transcription

Low-level transcription

No transcription

Strong transcription