Bacterial gene regulation Flashcards
molecular definitions of a gene?
1) the DNA sequence that is translated into a protein
2) the ORF and the control region of the gene including its promoter
dawkins definitions of a gene?
any portion of chromosomal material that potentially lasts for enough generations to serve as a unit of natural selection - Dawkins
flow of information in a cell?
DNA -> RNA -> protein
the central dogma describes…
…the flow of information in an organism
what do protein control points do?
removes rna / proteins which arent needed, for efficiency
in bacteria, transcription ansd translation is…
…coupled
where is bacteria in dna?
chromosomal DNA is free in the cytoplasm
because the chromosomal DNA is free in the cytoplasm translation can begin as soon as enough transcript is available for ribosome binding – this enables…
… gene regulation to be fast
the expression of genes is regulated at several levels which are?
1) transcriptional control
2) post-transcriptional control
3) translational control
4) post-translational control
Function of transcriptional control?
control of production of the mRNA transcript – requires promoters/operators (DNA) and activators/repressors (proteins) e.g. lac operon
Whats the function of post-transcriptional control?
mRNA stability – how long is the mRNA available for ribosomes to translate? e.g. Crp-cAMP control by RNaseL
whats the function of translational control?
inhibition of ribosome binding – e.g. threonyl tRNA synthetase (ThrS) binds to thrS mRNA inhibiting ribosome binding
Whats the function of post translational control?
activation or inactivation of an already translated protein
a) protein stability/degradation rates
b) post-translational modification – addition of phosphate groups (phosphorylation)
Whats the most important level of gene expression in bacteria?
In bactiera, the mos timportant level of control is transcriptional control
How many regions in each gene?
3
What are the three regions of a gene?
1) 5’ promoter, attracts RNA polymerase
2) transcribed sequence (transcript) or RNA coding sequence
3) 3’ terminator, signals the stop point
Whats the function of the RNA-coding sequence?
rna coding sequence starting with aug and ending with stop codon
Function of the +1 on a gene?
where transcription starts.
Whats the terminator of a gene?
stops transcribing and moves off gene.
Function of promoter?
The function of a promoter in transcription is to control when and where a gene is expressed in an organism
What is RNA polymerase made up of?
RNA polymerase made up of subunits, transcribes all genes.
unlike in eukaryotes only one bacterial RNA polymerase transcribes…
… all genes – coding genes, rRNA and tRNA genes
What does RNA polymerase enzyme consist of?
the enzyme consists of the core alpha and beta subunits
What are the core alpha and beta subunits of RNA polymerase involved in?
formation of RNA via phosphodiester bonds adding nucleotides to a growing RNA chain.
The rna polymerase complex is unable to…
… bind to DNA itself in order to initiate transcription
how does the rna polymerase complex bind to dna?
binding takes place via sigma (σ) factors
differential binding of sigma factors represents the most…
… general part of gene regulation in bacteria
control of transcription in bacteria requires…
… the presence of a promoter just upstream of the transcription initiation site
control of transcription in bacteria requires the presence of a promoter just upstream of the transcription initiation site – conservation occurs in…
… the promoter as these are DNA sequences that interact with proteins that are required to initiate transcription – sigma factors
Whats a sigma factor?
proteins that are required to initiate transcription
What are sigma factors required for?
transcription
different sigma factors control…
… different sets of genes.
How many sigma factors in E. coli?
in E. coli there are 7 different sigma factors each responsible for transcription initiation of different regulons
How does a sigma factor work?
1) Sigma factor recognises and binds to -35 and -10 sequences
2) Then sigma factor changes its shape so it can bind to rna polymerase
3) So then it known where to bind on a promoter and start transcription.
sigma factors bind to …
… DNA
the grooves within a double stranded DNA molecule are able to …
… interact with amino acids on the surface of a protein due to H-bonds and Van der Waals forces
the grooves within a double stranded DNA molecule are able to interact with amino acids on the surface of a protein due to H-bonds and Van der Waals forces
this provides the opportunity for proteins to …
… interact and importantly the interactions are sequence specific – hence the core promoter sequence being conserved!
Sigma factor changing shape of protein allows …
… the dna to interaction with it
Sigma factor changing shape of protein allows the dna to interaction with it. Because …
… Because the dna fits to the changed shape of the protein.
Sigma factor knows where to go
sigma factors are released on …
… initiation
initiation of transcription causes …
… the sigma factor to be released and RNA polymerase processes along the template strand.
Bound sigma factor recruiting …
… rna polymerase to the correct position