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Regulation of Gene Expression • Chromosomal Map begins at OriC; units of minutes. – Only structural genes for enzymes are shown here. – Their control regions (promoter and operator) determine transcription. – The complete organizational unit is an operon. • Transcriptional regulation: – Negative Control by Repressors • Repression • Induction – Positive Control by Activators – Attenuation (involves translation) Transcriptional Regulation by Repression • Regulatory protein (repressor) is encoded on a gene outside and away from the operon it regulates. • Active repressor binds operator region; RNA Polymerase blocked = negative control. • Repressor becomes active by a corepressor. • Corepressor is often an endproduct of pathway enzymes encoded on the operon. Transcriptional Regulation by Induction • Active repressor binds operator region; RNA Polymerase blocked = negative control. • Gene transcribed when inducer molecule is present; binds and inactivates repressor (release from operator). • Inducers are typically substrate for a pathway enzyme encoded on the operon (e.g. allolactose for the lac operon) Lactose Catabolism (lac) Operon Doesn’t work if glucose is available! Why? Transcriptional Regulation by Catabolic Activator Protein (CAP) • CAP = cyclic AMP receptor protein (CRP). • Active CAP binds promotor and allows transcription to proceed = positive control. • Activation of CAP requires build-up of cAMP to bind to CAP. • cAMP builds-up in cells not producing enough ATP due to lack of glucose availability. • The lac operon requires both lactose and cAMP. lac Operon in Action (diauxic growth) Separate cultures • PEP-PTS at high glucose uptake lowers adenyl cyclase activity; low cAMP; CAP inactive. • Exhaustion of glucose increases cAMP, activating CAP; repressor is inactivated; lac operon transcribed! Together Tryptophan (Trp) Operon (Trp synthesis (anabolic); regulated by repression and attenuation.) Transcriptional Regulation by Attenuation • In addition to a promotor and operator, the operon has a leader sequence with two pairs of selfcomplementing sequence sections (#1&2 and #3&4). The first pair is in what is called the leader peptide gene. • The second pair (#3&4) is part of a Rho-independent terminator region upstream of any structural genes; called an attenuator. Trp high. • Prevention of the first pair complementing will result in a hybrid complement of first and second pair (sections #2 and #3). Trp low. Transcriptional Regulation by Attenuation • Attenuation of transcription results when the attenuator hairpin can form. • It forms when there is no translation of leader sequence mRNA & when there is ample trp-tRNA. • Absence of trp-tRNA causes ribosome to stall, blocking section #1; hybrid forms. • No attenuation hairpin; RNA polymerase proceeds to transcribe genes. 1) No Translation; No genes transcribed! 2) Trp & trp-tRNA available 3) Trp & trp-tRNA absent