Green = lac repressor
Red = DNA, operator region
Blue = part of the operator (not a hairpin loop, its double stranded DNA)
Repressor = a dimer and when it binds to the operator, it creates a loop to make it impossible for polymerase to get through
In the lac operon, the beta galactoside bond is hydrolyzed by galactosidase either into glucose and galactose or by flipping the bond it is converted into allolactose
***Inducible (normal –> off until turned on)
- lacI – makes repressor; inducible
- lacZ – always off; no activity
- lacY – permease (transports lactose into cells)
NEGATIVE control because binding that protein reduces expression (negative has nothing to do with lactose or enzyme expression, just what happens when the protein binds to DNA)
Epigenetics – that DNA can learn from its environment, genome can respond to its environment
CAP binding
Binding site is upstream of the promoter – diamer
- Binding of CAP bends helix in a way that makes promoter more accessible (and stable) to RNA polymerase –> Therefore expression goes UP
- Only happens with CAP/cAMP complex
Metabolism of glucose results in decreasing cAMP (as metabolism of glucose increases, concentration of cAMP decreases – CAP binding drops – lac operon expression drops)
- Many operons are also under regulation of CAP
Normally:
CAP binds = expression goes up
Doesn’t bind (presence of glucose) = expression goes down
REGULON –> many genes control by same binding proteins
- Lactose present; glucose low/absent
- Lactose present; glucose present
Mutations
When glucose is added, nothing happens
When lactose is added, galactosidase expression increases
WHERE IS MUTATION??
Assumption that mutations are bad and cause decrease in function – Not TRUE, mutations can make things better
Mutation causes loss of function in CAP – mutation might be in CAP binding site, promoter, gene, etc (could be several mutations, but how can you tell the difference?): operon remains inducible, expression rises in presence of lactose but not repressed in presence of glucose cause CAP cannot bind
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