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- W2182785170 abstract "We report a modified synthesis for5-mercapto-UTP (5-SH-UTP) anditsuse foranalysis ofprotein-RNA interactions utilizing Escherichia coilandT7 RNA polymerases andyeastRNA polymerases IandIll. 5-SH-UTP didnotaffect transcriptional pausing, Rhoindependent termination orrecognition oftheE.coIi transcription complex byNusA.RNAcontaining 5-SHUMPdidnotcrosslink topolymerase whenirradiation was at302or 337nm. Transcription complexes containing RNA substituted with5-SH-UMPwere post-transcripti onally modified toattach aphotocrosslinking grouptothiol-tagged nucleotides intheRNAon thesurface ofthepolymerase orfreeinsolution. The pK,for5-SH-UTP was determined tobe 5.6,so modification ofthethiol groups intheRNA with p-azidophenacyl bromide couldbecarried outatpH7. Addition ofthetranscription termination factor Rho,a RNAbinding protein, toE.colitranscript ion complexes resulted inRNAcrosslinking toRhoandtotheI8and 5'subunits ofpolymerase. Using5-SH-UTP, one can distinguish RNA binding domains on thesurface of RNApolymerases orother RNAbinding proteins from thoseburied within theprotein." @default.
- W2182785170 created "2016-06-24" @default.
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- W2182785170 date "1995-01-01" @default.
- W2182785170 modified "2023-09-29" @default.
- W2182785170 title "Preparation ofprobe-modified RNAwith 5-mercapto-UTP foranalysis ofprotein-RNA" @default.
- W2182785170 hasPublicationYear "1995" @default.
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