THE lac OPERON

The control of gene expression

Each cell in the human contains all the genetic material for the growth and development of a human Some of these genes will be need to be expressed all the time Why? Which ones? These are the genes that are involved in of vital biochemical processes such as respiration Other genes are not expressed all the time Why? Which ones? They are switched on and off as needed What mechanism(s) could do this?

Prokaryotic Gene Regulation Operons

An operon is a group of genes that have a single regulator so that they are transcribed at the same time. They usually control an important biochemical process. They are only found in prokaryotes.

Jacob, Monod & Lwoff

NobelPrize.org

The lac Operon

The lac operon consists of three genes each involved in processing the sugar lactose One of them is the gene for the enzyme galactosidase This enzyme hydrolyses lactose into glucose and galactose The other genes are for producing the enzymes: lactose permease (transports lactose into cell) and transacetylase (function not known)
http://www.biocourse.com/ui/swf/iLabs/lac_operon.swf

http://www.biocourse.com/ui/swf/iLa bs/lac_operon.swf

Adapting to the environment

E. coli can use either glucose or lactose What is the preferred substrate? Suggest why Glucose is preferred because lactose needs to be hydrolysed (digested) before it can be respired

Four situations are possible

When glucose is present and lactose is absent the E. coli does not produce -galactosidase.
When glucose is present and lactose is present the E. coli does not produce -galactosidase. When glucose is absent and lactose is absent the E. coli does not produce -galactosidase. When glucose is absent and lactose is present the E. coli does produce -galactosidase

The control of the lac operon

The operator site

located between the promoter and the first structural gene this is the on/off switch
where the lac repressor protein binds the physical shape of the lac repressor blocks the path of RNA polymerase

lac repressor gene

Also called regulator gene this gene synthesizes a lac repressor protein the lac repressor protein binds to the operator

promoter site
next to the operator site where RNA polymerase binds

Lac operon animations http://www.youtube.com/watch?v=oBwtxdI1zvk simple introduction http://highered.mcgrawhill.com/olc/dl/120080/bio27.swf combination of switches http://www.sumanasinc.com/webcontent/animati ons/content/lacoperon.html simpler http://highered.mcgrawhill.com/sites/0072556781/student_view0/chapte r12/animation_quiz_4.html most complex shows how genes are expressed when glucose / lactose present / absent best to watch at the end!

1. When lactose is absent

A repressor protein is continuously synthesised. It binds to a sequence of DNA just in front of the structural genes of the lac operon, on the Operator site The repressor protein blocks the Promoter site where the RNA polymerase settles before it starts transcribing So transcription is switched off

2. When lactose is present

A small amount of a sugar allolactose is formed within the bacterial cell. This fits onto the repressor protein at another active site (allosteric site) This causes the repressor protein to change its shape (a conformational change). It can no longer sit on the operator site. RNA polymerase can now reach its promoter site Transcription is switched on

3. When both glucose and lactose are present

This explains how the lac operon is transcribed only when lactose is present. BUT.. this does not explain why the operon is not transcribed when both glucose and lactose are present. Another regulatory site called the CAP binding site is important here

First consider when glucose is absent

Glucose is absent and lactose is present

Glucose is present

Glucose is present
cAMP levels are low when glucose is present So cAMP is not available to bind to the CAP protein which cannot bind to the CAP binding site RNA polymerase cannot bind to the promoter Structural genes to metabolise glucose are not transcribed Glucose is metabolised in preference to lactose

Lac operon showing regulatory sites

Extension: Gene Control in Eukaryotes

1. Chromatin Structure 2. Epigenetic Control 3. Transcriptional Initiation 4. Transcript Processing and Modification: splicing and capping of mRNA 5. RNA Transport 6. Transcript Stability 7. Translational Initiation 8. Small RNAs and Control of Transcript Levels 9. Post-Translational Modification: 10. Protein Transport 11. Control of Protein Stability