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    Lac Operon Key Points

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    amirtha03akilan06

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    Lac Operon Key Points

    Uploaded by

    amirtha03akilan06
    14 views12 pages

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    lac operon Key points

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    14 views12 pages

    Lac Operon Key Points

    Uploaded by

    amirtha03akilan06

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    of 12

    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
    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.

    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?]

    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 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 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
    CAP Repressor Level of
    Glucose Lactose binds binds transcription
    + - - + No transcription

    Low-level
    + + - - transcription

    - - + + No transcription

    - + + - Strong transcription

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