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Inducible transcription factors that 2 respond to

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Unformatted text preview: TTR transcription transcription • TTR promoter: model for cooperative assembly of basal transcription apparatus in hepatocytes hepatocytes RESPONSE ELEMENTS: COMMON REGULATORY CONTROL REGULATORY • Groups of genes under Groups • common regulation often share specific factorshare binding sequences (1) • RESPONSE ELEMENTS: RESPONSE bound by inducible transcription factors that (2) respond to signals respond • Often found in promoter & Often enhancer regions enhancer • Not found at fixed Not positions but in promoters usually <200bp from start site site eg. COMMON COMMON RESPONSE ELEMENTS: ELEMENTS: HSE: heat shock HSE: response element - in promoters & bound by HSTF HSTF GRE: glucocorticoid GRE: response element enhancer recognized by steroid receptor by RESPONSE ELEMENTS: COMMON REGULATORY CONTROL REGULATORY -heat shock response that is controlled thorugh HSE underscores differences between prokaryotic and eukaryotic gene expression -heat shock response in bacteria mediated by production of new σ factor that enables RNAP to recognize special promoters of heat shock genes -eukaryotic heat shock genes are activated by binding of HSTF to HSE sequences in their promoters -heat shock apparatus well conserved among eukaryotes, for example, Drosophila heat shock genes can be activated in mammals and sea urchins METALLOTHIONEIN GENE: MULTIPLE RESPONSE ELEMENTS Weaver 2002 Molecular Biology Ed. 2 • • • Complex interactions of enhancers permits regulation of eukaryotic gene Complex expression in response to wide-range of conditions expression eg. Metallothionein gene regulation: constitutive factors (Sp1/GC boxes) + Metallothionein response elements (GRE - gluccocorticoid, MRE - inductive response to heavy metal ions, TRE (inside one BLE)- AP1 binding triggered by phorbol esters) esters) metallothionein protein protects cells from excess concentrations of heavy metallothionein metals by binding and removing them from the cell metals USE OF CHROMATIN IMMUNOPRECIPITATION (ChIP) TO DETERMINE TRANSCRIPTION FACTOR BINDING SITES DETERMINE -if you have an antibody against a transcription factor, ChIP can be used to determine where in a gene of interest that transcription factor binds -PCR primers targeting segments of the gene of interest are used to determine which sequences are present in the immunoprecipitation USE OF CHROMATIN IMMUNOPRECIPITATION (ChIP) TO DETERMINE TRANSCRIPTION FACTOR BINDING SITES FACTOR -genome-wide analysis of transcription factor binding sites can be done using microarrays (ChIP-chip) or high-throughput sequencing (ChIP-seq) -sample results of ChIP-seq reveal variations in transcription factor binding patterns YEAST ONE-HYBRID CAN BE USED TO FIND TRANSCRIPTION FACTOR(S) BINDING...
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