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lecture8 - RNA synthesis Gene Transcription The E.coli RNA...

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801 RNA synthesis Gene Transcription The E.coli RNA polymerase
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Gene mRNA Translation at ribosome Protein Transcription Transcription: DNA -> RNA
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mRNA: messenger RNAs tRNA: transfer RNAs rRNA: ribosomal RNAs snRNA: small nuclear RNAs miRNA: micro RNAs gRNAs: guide RNAs all RNAs derive from genes! all RNAs are synthesized by transcription!
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(RNA) n+1 + PP i 2P i (RNA) n + NTP H 2 O The Reaction Initiation Elongation Termination DNA + RNA Three phases
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E.coli RNA polymerase: Holoenzyme initiates: α 2 ββ σ Core Enzyme elongates: α 2 ββ Zn 2+ Mg 2+
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“hand” shape with channel that fits B-DNA thumb may close channel T7 RNA polymerase: <100kd E. coli RNA polymerase: >400kd
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RNA Polymerase from Taq ß-subunit ß’-subunit α -subunit α -subunit Mg 2+ Zn 2+
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MgCl 2+
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Gene Transcription +1 gene
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Regulation of Transcription (in E.coli)
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+1 Gene structure in E.coli: Operons - transcription units that can be independently regulated monocistronic RNA or polycistronic RNA +1
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upstream downstream 5’ 3’ +1 gene transcription start site (TSS) is a reference point
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Promoter Sequences in E.coli for Initiation sense strand binding by RNA holoenzyme: partial DNAse1 digestion evolutionary conservation: consensus sequences functional test: mutation affects gene transcription rate How do we know these sequences are important?
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Promoters: locations where proteins are bound DNAse 1 footprinting : to identify protein-DNA interaction sites
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DNAse1 Footprinting
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1. End-label DNA fragment.
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lecture8 - RNA synthesis Gene Transcription The E.coli RNA...

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