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Chapter 17
Chapter 17
Chapter 19:
Through evolutionary processes, organisms have developed ways to compensate for environmental changes.
The regulator substance is called an inducer, which are members of a class of small molecules = effectors
Controlling site is near protein coding sequence. The addition of inducer leads to induction.
When Lactose is only sugar, three enzymes are synthesized 1) B-galactosidase 2) Lactose permease 3) Trans acetylase (function poorly understood)
All 3 genes are clustered on the genome and are transcribed onto a single mRNA called a polygenic mRNA or a polycistronic mRNA
Nonsense mutants (chain terminating mutant) were used to determine that all 3 genes were on the same mRNA.
Jacob & Monods Operon Model for the regulation of the lac genes
Operon is a cluster of genes, the expression of which are regulated together by operator regulator protein interactions, plus the operator region itself and the promoter.
In E. coli, the lac operon is under negative, positive and inducible control
Lac I+ gene encodes lac repressor protein made constitutively, which will bind operator region of the lac operon. Few repressors are present in cell since promoter is relatively weak.
Absence of lactose
Lac operon is under negative control: There is a low level of lac gene expression because the repressor binds and unbinds allowing for low amounts of protein such as B-galactosidose and permease to be generated
Presence of lactose
Inducible Control
If in the presence of lactose, the above B-galactosidose produces inducer molecules, allolactose, which is the inducer.
Without inducer
With inducer
Mutation in Lac I gene, which generates a mutant repressor that cannot bind to the operator
Without inducer
Mutation in Lac I gene, which generates a mutant repressor that cannot bind to the operator
With inducer
Dominant Effect Mutation in Lac I that cannot bind Inducer but can bind the operator
**This data convinced many scientist (at the time) that all genes were under negative control due to the binding of a repressor. **
Shine-Dalgarno sequence
Tryptophan Operon
All necessary amino acids may not be present in a growth medium. If a specific amino acid is missing, the bacteria has certain operons that enable the bacterial cell to manufacture that amino acid.
For example the Tryptophan Operon
Two mechanisms of regulations for tryptophan operon # 2 Attenuation Control Regulatory Leader region Determines if initiated transcripts include other structural genes or not.
The biosynthetic pathway is catalyzed by a specific enzyme (at each step) which is coded by a specific gene or genes. Presence of tryptophan in medium keeps operon turned off
# 2 Attenuation Control
Absence of tryptophan or in the presence of low amounts of tryptophan: Under severe tryptophan starvation gene activity at maximum all long transcripts
Greater the amount of tryptophan the greater the number of short transcripts Attenuation controls = > terminates transcription producing short transcripts
mRNA
Position of ribosome on leader transcript determines if transcription is terminated or not. If starved for trytophan = lack trp-tRNA If no trp-tRNA, ribosome stalls at trp codons. With Ribosome on region one, the 1-2 loop cant form. So, the 2-3 loop for antitermination forms. Thus region 3 cant pair with region 4 and the RNA polymerase can now continues.
++ trptophan
In the lac operon, allolactose would bind the repressor to remove it from the operator so that the Polymerase could bind the operator and start transcription. In the ara operon, two molecules of AraC protein bind and act as a bridge from the operator (araO 2 ) And to the promoter region ara I1 which creates a loop that prevents the binding of CAP-cAMP. With the addition of arabionose, the arabionose bound AraC protein is allosterically modified to bind to ara I2, which allows CAP-cAMP to bind the CAP site and positive regulate gene expression occurs.
However, for the ara operon to function, glucose can not be Present. If present, it will eliminate cAMP and focus on the Utilization of glucose
**Positive regulation of activators is now known to occur in a Variety of prokaryotic systems and in all eukaryotes.
1) 2)
Linear chromosome is circularized in host Transcription begins at PL & PR PL promoter for left early operon PR promoter for right early operon
Depends on a genetic switch, which involves competition between the products of the CI gene (the repressor) and the Cro gene (the Cro protein) regulator of CI gene.
Left Right
Important info
cI gene Cro gene N gene N = protein is the antiterminator that allows RNA transcription past transcription terminator signals. Lysogeny Lytic cII protein stimulates synthesis of cI repressor which competes with Cro protein. Decision on which pathway is taken is determined by the amount of repressor or Cro protein that is bound to PR or OR region.
cI protein
Integration of
Overheads 1, 2 and 3