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Lac Operon

1. The lac operon controls genes for lactose digestion in E. coli. 2. When lactose is present, it binds to the repressor protein which changes shape so it cannot bind to the operator region. This allows RNA polymerase to transcribe the structural genes encoding lactose permease and beta-galactosidase. 3. When no lactose is present, the repressor protein binds to the operator region and blocks RNA polymerase from transcribing the structural genes.

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172 views20 pages

Lac Operon

1. The lac operon controls genes for lactose digestion in E. coli. 2. When lactose is present, it binds to the repressor protein which changes shape so it cannot bind to the operator region. This allows RNA polymerase to transcribe the structural genes encoding lactose permease and beta-galactosidase. 3. When no lactose is present, the repressor protein binds to the operator region and blocks RNA polymerase from transcribing the structural genes.

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Write three different sentences using the following words -


1. Mutation
2. Chloride ions
3. Totipotent
Regulation of genes
Transcription factors and lac operon
Switching on genes
• We have learnt that cells become specialised when active genes are
transcribed into mRNA and then translated into proteins.
• These proteins modify the cell and determine the role of the cell.
• So what controls the expression of a gene?
Switching on genes
• Last lesson you learnt about epigenetic changes and how methylation
of DNA and acetylation of histone proteins can switch genes on/off on
eukaryotic DNA.

• What about prokaryotic DNA?


Transcription factors
• In prokaryotes, control of gene expression involves transcription
factors binding to operons.

• An operon is a length of DNA made from structural genes and control


sites.
Lac Operon in E.coli

• http://www.youtube.com/watch?gl=GB&v=oBwtxdI1zvk
Lactose utilisation in E.coli – the lac operon

 Requires a transport protein for the uptake of lactose across


the cell membrane
 Requires an enzyme to break lactose down into glucose and
galactose
 It would be wasteful to produce these in the absence of lactose

The expression of
these genes is
induced by the
presence of lactose
Regulatory gene Structural gene – lac operon

p lacI RNA
lacP
pol

Promoter Promoter
region region

The structural genes encode for useful proteins involved in the


uptake(transport protein) and utilisation of lactose(enzyme)

The regulatory gene encodes a protein which controls when the


structural gene is expressed (an ON/OFF switch) coding for
an activator or repressor
RNA polymerase binds to the promoter region to initiate
transcription of gene
Regulatory gene Structural gene – lac operon

lacI lacO lacZ lacY lacA

lacZ encodes  - galactosidase


- an enzyme which hydrolyses lactose to
produce glucose and galactose

lacY encodes permease - proteins which transport


lactose into the cell

lacA encodes  - galactosidase transacetylase


- adds an acetyl group to  - galactosidase
(Nobody knows why!)
Regulatory gene Structural gene – lac operon

RNA
pol lacI lacO lacZ lacY lacA

transcription
1. The lacI is transcribed by RNA polymerase...

... and translated by ribosomes to produce a


transcription repressor protein
The repressor protein has two binding sites

- One site binds to lactose


- The other site bind to lacO – the operator
Lactose DNA region
binding binding
site site
Switching the Structural Genes OFF
Regulatory gene Structural gene – lac operon

lacI lacO lacZ lacY lacA

2. When lactose is ABSENT


RNA the repressor protein
pol binds to lacO

This blocks lacP so that RNA cannot bind to it


and the structural genes cannot be transcribed
Switching the Structural Genes ON
Regulatory gene Structural gene – lac operon

RNA lacO lacZ lacY lacA


lacI
pol

3. When lactose is PRESENT ... this changes the shape of


it binds to the lactose- the DNA binding site so...
binding site on the
... that it cannot bind to
repressor protein...
lacO

This allows RNA polymerase to bind to lacP so that the structural


genes can be transcribed
The lac operon
•As a result of the 2 enzymes being made, e.coli can now.....

•Take up lactose from its environment because lactose permease


acts as a transport protein.

•Using ß-galactosidase, lactose (disaccharide) can hydrolysed into


glucose & galactose

•E.coli can use these sugars for respiration

•E.coli is gaining energy from the lactose


Task
• Complete the fill in the gap exercise.
• Annotate the diagrams of lac operon that show what happens when
lactose is present and absent.
• Answer the questions
http://www.youtube.com/watch?gl=GB&v=oBwtxdI1zvk

Q.1 match the components with the functions.


Component Function
A. Structural Gene 1. Produces repressor protein
B. Regulatory gene 2. Binds to repressor
C. Promoter 3. Codes for lac enzymes
D. Operator 4. Binds to RNA polymerase

Q.2 What are the functions of:


•Repressor protein
•RNA polymerase
•Regulatory gene
Student task
•Get into groups of 3 or more

•Use the large A2 paper as a story board

•Each person is to cut out the parts of the lac operating system on
the handout provided

•Arrange the cut out parts to show the story of how the lac operon
works in both the absence and presence of lactose
regulatory
idea that makes , repressor protein / transcription factor
idea that product switches (structural / another) gene , on / off
structural -idea that makes , enzyme / polypeptide / protein ;
relationship between the 2
idea that regulatory gene , controls / affects , the expression of structural
gene ;

lactose has been , removed / digested / respired / broken down (by


bacteria) ;
to , lactic acid / lactate / other sugars ;
yogurt still a good source of , calcium / vitamins ;
lactose binds to repressor protein ;
changes , shape / structure (of protein) ;
removes it from / stops it binding to , operator ;
RNA polymerase binds to promoter ;
idea that (so that Z and Y) are , transcribed / mRNA made

Exam Question Jan 2011


The Lac Operon – Summary
Genes responsible for lactose digestion and their regulation.
Physical presence of lactose results in transcription of genes
to produce lactose permease and Beta-galactosidase.
A regulatory gene produces a repressor protein. When no
lactose is present, it joins with the operator region.
RNA polymerase joins to the promoter region. If the repressor
protein is attached to the operator region, the RNA
polymerase cannot move further along and transcript
structural genes to produce enzymes for lactose
digestion.
If lactose is present, it binds with the repressor protein and
causes a conformational change, so the repressor protein
can’t bind with the operator region.
RNA polymerase can travel along and the enzymes for lactose
digestion can be produced.

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