Control of gene expression Receptors and transcription factors Inducible expression of -galactosidase (lac operon) Structure of lac operator and repressor Positive and negative regulation Steroid hormone control of gene expression Interference RNA
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Control of gene expression Receptors and transcription factors Inducible expression of -galactosidase (lac operon) Structure of lac operator and.
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Control of gene expression
Receptors and transcription factors
Inducible expression of -galactosidase (lac operon)
Structure of lac operator and repressor
Positive and negative regulation
Steroid hormone control of gene expression
Interference RNA
Control of gene expression
DNA
mRNA
proteins
receptors
transciption factors
ENVIRONMENT
CELL
Receptors are proteins which trigger
intracellular chemical signaling in response to
an external stimulus
Transcription factors are proteins which
control mRNA synthesis
-galactosidase is an inducible enzyme
-ga
lact
osid
ase
+ inducer
- inducer
temps
An inducer is a molecule which modulates the level of expression of a protein
-galactosidase allows Escherichia coli to grow using lactose as its only carbon source
The lac operon : a set of co-regulated genes
-galactosidase (z) permease (y) transacetylase (a)E.coli DNA
ATG STOP ATG STOP ATG STOP
Mutants
defective
constitutive
non-inducible
z-, y-, a-
i-
Complementation
Genomic DNA z+, y+, a+i-
F’ episome z-, y+, a+i+
-galactosidase expression
E. coli i-z+
E. coli i+z-
E.coli i-z+/F’i+z-
-lactose + lactose+ +- -- +
Regulation of -galactosidase expression by the lac repressor
p repressor
In the presence of the inducer
In the absence of the inducer
p repressor
The inducer binds to the repressor, which prevents it from interacting with the operator
p o -galactosidase
p o -galactosidase
Binding of the repressor to the operator prevents -galatosidase transcription
Structure of the lac operator
The lac repressor protects the lac operator from digestion by pancreatic desoxyribonuclease
The lac operator has a symmetric nucleotide sequence
Some repressor structures
Trp repressor bound to its DNA operator
phage repressor bound to its DNA operator
Interaction between transcription factors and DNA
Specific interaction : some amino-acids of the transcription factor directly bind to the outside of DNA bases
Non-specific interaction : Some amino acids of the transcription factors bind to the DNA polyphosphate chains
Asparagine
CH 2
CO
NH 2
CH 2
Structure of the lac repressor
Dimerization domain
Inducer binding domain
Operator binding domain
The lac repressor changes its conformation upon inducer binding
operator
IPTG
Repressor structure in the absence of IPTG
Repressor structure in the presence of IPTG
Dimerization of the lac repressor increases the affinity for the operator