Journal
MOLECULAR CELL
Volume 79, Issue 5, Pages 797-+Publisher
CELL PRESS
DOI: 10.1016/j.molcel.2020.07.006
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Funding
- NIH [GM124976, GM041376, GM118059]
- Helen Hay Whitney Foundation
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Pausing by RNA polymerase (RNAP) during transcription elongation, in which a translocating RNAP uses a stepping'' mechanism, has been studied extensively, but pausing by RNAP during initial transcription, in which a promoter-anchored RNAP uses a scrunching'' mechanism, has not. We report a method that directly defines the RNAP-active-center position relative to DNA with single-nucleotide resolution (XACT-seq; cross-link-between-active-center-and-template sequencing''). We apply this method to detect and quantify pausing in initial transcription at 4(11) (similar to 4,000,000) promoter sequences in vivo in Escherichia coli. The results show initial-transcription pausing can occur in each nucleotide addition during initial transcription, particularly the first 4 to 5 nucleotide additions. The results further show initial-transcription pausing occurs at sequences that resemble the consensus sequence element for transcription-elongation pausing. Our findings define the positional and sequence determinants for initial-transcription pausing and establish initial-transcription pausing is hard coded by sequence elements similar to those for transcription-elongation pausing.
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