REGULATION OF GENE EXPRESSION Flashcards
What is the transcription of many genes controlled by?
Activators and repressors
What is the activity of activators and repressors controlled by?
Inducer or codepressors
Why are all the genes not expressed at once?
In prokaryotes- different genes are expressed in response to different environments e.g. food present such as lactose
- Also don’t waste energy expressing genes which don’t need to be used
What do eukaryotic genes need to express?
DIFFERENT FACTORS as they need to be able to differentiate into different cell types
What is the ground state for bacterial genes?
They are always ON unless inactivated
What is the ground state for eukaryotic genes?
They are always OFF unless activated
- They need another 10-20 proteins to activate RNA polymerase transcription
Why are eukaryotic genes hard to access?
- Because they are wrapped tight in chromosomes (histone)
What does regulation at the transcription level do?
- Regulate how much mRNA is made from a gene
- Without mRNA final protein can’t be expressed
- Most common level of regulation
What does regulation at the level of translation do?
- Regulates how much active protein is made from mRNA transcript
What determines where RNA polymerase starts transcription?
- Promoter sequences e.g. TATAAT or TTGACA
What is negative regulation?
- When a regulatory protein is used to stop transcription of a gene(turns promoter OFF)
- Repressor
What is the RNA polymerase in bacteria recruited to the promoter by?
- By sigma factor
When does negative regulator need to be inactivated?
- When genes need to be turned on in response to a stimulus
What is positive regulation?
- Activator
- Regulatory protein required before the gene is efficiently transcribed (turn promoter ON)
When does a positive promoter need to be activated?
- When genes need to be turned on in response to a stimulus
What will slightly different promoter sequences be recognised by/how is RNA polynmerase (prok) and thounsands of genes explained?
- Different sigma factors recognising alternative promoter sequences
e. g sigma 32 (genes induced by heat shock) so levels of this then rise when heat shock happens
What is the place where the repressor binds called?
- The Operator site
- (promoter and operator overlap)
How do repressor proteins stop transcription?
- By repressor binding over the promoter sequence so RNA polymerase can’t bind to the DNA
What is sometimes required for the repressor to bind(negative regulation of transcription)?
- A corepressor
Repressor + corepressor=
ACTIVE site for binding
What is the region between -35 and +1?
- The promoter region (overlaps with the operator region)
What section on the DNA is the operator region?
-5 to +21
What happens to the repressor in negative regulation to allow for transcription?
It binds to the inducer (no longer binding to the operator) to allow for transcription to occur
How is repression relieved?
By an inducer
What are the three steps in allowing transcription in negative regulation?
- Repressor binds to the operator site and stops transcription
- When inducer is present it binds to repressor and repressor falls off DNA
- RNA polyemerase can now access promoter and direct transcription
What is the repressor and inducer for lactose respectively?
- LacI (repressor) and allo-lactose (inducer)
What occurs in positive regulation of transcription?
Activator can only bind to DNA in presence of inducer (inducer interacts with the activator allowing it to bind to DNA)
What are the two steps in positive gene expression to allow for activation?
- Activator must bind to promoter to allow transcription to begin
- Activator can only bind to DNA in presence of inducer
Where is the binding site for activator proteins?
- Upstream of the promoter
- needs inducer to bind
e. .g maltose is an inducer (binds to activator protein–> changes conformation–> can then bind to activator site upstream of promoter.)
What are allosteric proteins?
Proteins that exist in two different conformational forms (shapes)
- Change conformation when binding to the inducer
What are examples of allosteric proteins?
- Transcriptional repressors and activators
What does the change in conformation of allosteric proteins alter?
- The ability to bind DNA
What are the two different binding sites that the allosteric proteins have?
- One for binding inducer
- One for binding DNA
What is the negative regulation of the Lac operon controlled by?
- Lactose repressor (product of LacI)
- Inducers are low MW beta-galactosides (allo-lactose)
What is the positive regulation of the lac operon controlled by?
- CAP protein
- Inducer is cAMP ( which responds to glucose)
What does the lac operon sense?
- not the actual glucose concentration BUT the cAMP levels
- Levels of cAMP and positive regulator CAP (catabolite activator protein)- catabolite repression
What do glucose levels regulate?
cAMP levels
Will the lac operon be expressed when glucose is present?
NO!!! It won’t because glucose has first preference
What does catabolite repression allow for?
- The fast adaptation of bacteria to a preffered carbon source
What happens in levels of high glucose?
- No cAMP produced
What happens in levels of low glucose?
- cAMP produced from ATP
How does the cAMP-CAP complex activate transcription in positive regulation?
- Complex binds to promoter and activates transcription
-
What effect does CAP have on DNA?
- It ‘bends’ the DNA (tortional pressure separates the DNA)
What does bent DNA allow for in positive regulation?
For RNA polymerase to access the promoter more efficiently
When does CAP activate transcription efficiently?
- In the absence of the lac repressor
When is there high levels of the lac operon expression?
- When glucose is absent or lactose is present
What happens when there is glucose present (cAMP low) and no lactose?
- Repressor binds (CAP doesn’t bind)
- NO EXPRESSION OF LAC mRNA
What happens when glucose is present (cAMP low), and lactose present?
- Repressor doesn’t bind, CAP doesn’t bind
- LITTLE EXPRESSION OF LAC mRNA
What happens when glucose is absent (high cAMP) and lactose is present?
- The CAP binds (CAP cAMP complex)
- HIGH EXPRESSION OF LAC mRNA
The lac operon has both:
- Positive and negative regulation (Lac repressor and CAP activator)