Applied force reveals mechanistic and energetic details of transcription termination.

Abstract:

:Transcription termination by bacterial RNA polymerase (RNAP) occurs at sequences coding for a GC-rich RNA hairpin followed by a U-rich tract. We used single-molecule techniques to investigate the mechanism by which three representative terminators (his, t500, and tR2) destabilize the elongation complex (EC). For his and tR2 terminators, loads exerted to bias translocation did not affect termination efficiency (TE). However, the force-dependent kinetics of release and the force-dependent TE of a mutant imply a forward translocation mechanism for the t500 terminator. Tension on isolated U-tracts induced transcript release in a manner consistent with RNA:DNA hybrid shearing. We deduce that different mechanisms, involving hypertranslocation or shearing, operate at terminators with different U-tracts. Tension applied to RNA at terminators suggests that closure of the final 2-3 hairpin bases destabilizes the hybrid and that competing RNA structures modulate TE. We propose a quantitative, energetic model that predicts the behavior for these terminators and mutant variants.

journal_name

Cell

journal_title

Cell

authors

Larson MH,Greenleaf WJ,Landick R,Block SM

doi

10.1016/j.cell.2008.01.027

subject

Has Abstract

pub_date

2008-03-21 00:00:00

pages

971-82

issue

6

eissn

0092-8674

issn

1097-4172

pii

S0092-8674(08)00127-X

journal_volume

132

pub_type

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