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Interactions between RNA polymerase and the "core recognition element" counteract pausing.


ABSTRACT: Transcription elongation is interrupted by sequences that inhibit nucleotide addition and cause RNA polymerase (RNAP) to pause. Here, by use of native elongating transcript sequencing (NET-seq) and a variant of NET-seq that enables analysis of mutant RNAP derivatives in merodiploid cells (mNET-seq), we analyze transcriptional pausing genome-wide in vivo in Escherichia coli. We identify a consensus pause-inducing sequence element, G???Y??G(+1) (where -1 corresponds to the position of the RNA 3' end). We demonstrate that sequence-specific interactions between RNAP core enzyme and a core recognition element (CRE) that stabilize transcription initiation complexes also occur in transcription elongation complexes and facilitate pause read-through by stabilizing RNAP in a posttranslocated register. Our findings identify key sequence determinants of transcriptional pausing and establish that RNAP-CRE interactions modulate pausing.

SUBMITTER: Vvedenskaya IO 

PROVIDER: S-EPMC4277259 | biostudies-literature | 2014 Jun

REPOSITORIES: biostudies-literature

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Interactions between RNA polymerase and the "core recognition element" counteract pausing.

Vvedenskaya Irina O IO   Vahedian-Movahed Hanif H   Bird Jeremy G JG   Knoblauch Jared G JG   Goldman Seth R SR   Zhang Yu Y   Ebright Richard H RH   Nickels Bryce E BE  

Science (New York, N.Y.) 20140601 6189


Transcription elongation is interrupted by sequences that inhibit nucleotide addition and cause RNA polymerase (RNAP) to pause. Here, by use of native elongating transcript sequencing (NET-seq) and a variant of NET-seq that enables analysis of mutant RNAP derivatives in merodiploid cells (mNET-seq), we analyze transcriptional pausing genome-wide in vivo in Escherichia coli. We identify a consensus pause-inducing sequence element, G₋₁₀Y₋₁G(+1) (where -1 corresponds to the position of the RNA 3' e  ...[more]

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