Lecture 7 - Regulation of Gene Expression 05-22-12-1
The document summarizes gene expression regulation in prokaryotes. It discusses the central dogma of genetics, levels of gene regulation including transcriptional and translational levels, regulatory genes and elements, DNA binding proteins, prokaryotic gene and operon structure, and examples of negative and positive control including the lac and tryptophan operons. The lac operon is induced by lactose and shows both negative and positive control, while the tryptophan operon is negatively regulated.
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Lecture 7 - Regulation of Gene Expression 05-22-12-1
The document summarizes gene expression regulation in prokaryotes. It discusses the central dogma of genetics, levels of gene regulation including transcriptional and translational levels, regulatory genes and elements, DNA binding proteins, prokaryotic gene and operon structure, and examples of negative and positive control including the lac and tryptophan operons. The lac operon is induced by lactose and shows both negative and positive control, while the tryptophan operon is negatively regulated.
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Lecture 7 05-23-12 Chapter 16
Gene Expression Regulation in Prokaryotes
Central Dogma of Genetics
This is the simplified model
Two important processes: Transcription and translation
Levels of Gene Expression Regulation
Only in DNA level eukaryotic Transcriptional cells level Transcriptional activators mRNA processing RNA stability Translational level Rate of translation Post-translational Protein modification
Regulatory Genes and Regulatory Elements
Structural genes: encode proteins play a structural roles in the cell Regulatory genes: genes whose products, either RNA or protein, interact with DNA sequences of other structural genes and affect their transcription or translation. DNA binding proteins - regulatory proteins Regulatory elements: DNA sequences, where the proteins produced by regulatory genes recognize and binds. Regulation of gene expression Stimulate gene expression: positive control Inhibit gene expression: negative control
DNA Binding Proteins
DNA Binding Proteins: regulatory proteins that bind to DNA sequences and affect gene expression at transcriptional level. Discrete functional parts Domains Domains consist of 60-90aa and are responsible for binding to DNA Motifs: simple characteristic structure within binding domain, that can fit to the major groove of DNA Distinct types based on characteristic motifs
Prokaryotic Gene Structure
Bacterial genes with related functions are often clustered together with one Single promoter and transcribed together into a single mRNA Operon: a group of bacterial functional genes that are transcribed together plus the controlling sequences. Operon organization Regulator Gene encodes regulator protein (with its own promoter) controlling the transcription of other genes Promoter Initiate the structural gene expression Operator DNA sequence binds to regulator protein to regulate transcription between the promoter and the structural genes Structural genes: encodes functional proteins
Operon Structure
The operator is a DNA sequence at the border between the promoter and the structural genes
Negative and Positive Control
Two types of transcriptional control Negative Regulator protein repressor Binds to DNA inhibiting transcription Negative Inducible Negative Repressible Regulator protein activator Binds to DNA stimulating transcription Positive Inducible Positive Repressible
Positive
Several variations of the above
Inducible = transcription is normally off Repressible = transcription is normally on
Negative Inducible Operon
Normally, the regulator protein is bound to the operator and prevents transcription Inducer: a small molecule, which can bind to the repressor Repressor binds operator = no transcription
Negative Inducible Operon
An INDUCER must be present to bind to regulatory protein (repressor) and prevent its binding to the operator for transcription to occur Repressor cant bind = transcription
Negative Repressible Operons
Normally transcription is on, regulator protein (repressor) is inactive and not bind to the operator no repressor = no binding on operator = transcription
Negative Repressible Operons
Co-repressor binds to the repressor allowing it bind to the operator. Transcription stops Product (co-repressor) present = binding = no transcription
Negative Repressible Operons
REMEMBER Negative control regulator protein = repressor, represses transcription. Inducible: normally no transcription and can be turned on Repressible: transcript normally is on and can be turned off
Positive Inducible Operons
Transcription normally is off. The regulator protein can be activated to bind to DNA (site other than operator) and activates transcription
Positive Repressible Operons
Normally, the regulator protein binds to DNA and activates transcription. Product inactivate the regulator protein and transcription turn off.
lac Operon of E. coli
Lactose: a type of sugar, an energy source for bacteria, E. coli Lactose operon: controls metabolism of the lactose in cell - lacI: regulator gene with it own promoter repressor - lacP: common promoter for 3 structural genes - lacO: operator where the repressor binds - 3 adjacent structural genes: lacZ encodes -galactosidase, which converts lactose into glucose and galactose for consumption or converts lactose in to allolactose, which regulates lactose metabolism. lacY encodes Permease allowing lactose to enter the cell. lacA encodes Transacetylase with unknown function
lac Operon of E. coli
When lactose or allolactose is present in the medium, where the bacteria grows, all 3 genes are expressed to metabolize lactose. inducible Example of negative inducible operon How are the genes activity induced, and repressed when lactose is absent??
lac Operon- Negative Control
lac Operon- Negative control
lac Operon- Positive Control
lac Operon also shows positive regulation lacZ gene codes for b-galactosidase which cleaves lactose into glucose, which the cell utilizes and prefers, and galactose when glucose is present at high levels, regardless if lactose is present, no transcription of lac operon Because b-galactosidase is not needed until the cell uses up the glucose, and then when lactose is present, lac operon gets transcribed. CAP mechanism controls the repression of the lac operon in the presence of glucose. Results in efficient energy use in cell. No need to metabolize lactose in the presence of glucose
lac Operon- Positive Control
CAP - Catabolite Activating Protein (CAP) CAP binds to cyclic adenosine monophospate (cAMP) cAMP alters the structure of CAP cAMP-CAP complex must bind to the promoter region for RNA Polymerase to bind for transcription of the structural genes. Glucose regulate cAMP High glucose = low cAMP, cAMP-CAP complex is not formed RNA polymerase cant bind to transcribe lac operon Low glucose = high cAMP RNA polymerase binds to the promoter for transcription
lac Operon- Positive Control
lac Operon- Positive Control
Tryptophan trp Operon
trp operon: Negative repressible gene system 5 structural genes Produce enzymes that synthesize tryptophan Upstream is the trp promoter Regulator gene (trpR) Product = regulator protein repressor Low tryptophan levels Repressor cant bind to DNA operator RNA polymerase binds DNA transcription occurs - tryptophan synthesis Tryptophan present It binds to the repressor binds to DNA operator Repress RNA polymerase binding no transcription