Studying transcription initiation by RNA polymerase with diffusion-based single-molecule fluorescence

Yazan Alhadid, Sang Yoon Chung, Eitan Lerner, Dylan J. Taatjes, Sergei Borukhov, Shimon Weiss*

*Corresponding author for this work

Research output: Contribution to journalReview articlepeer-review

8 Scopus citations

Abstract

Over the past decade, fluorescence-based single-molecule studies significantly contributed to characterizing the mechanism of RNA polymerase at different steps in transcription, especially in transcription initiation. Transcription by bacterial DNA-dependent RNA polymerase is a multistep process that uses genomic DNA to synthesize complementary RNA molecules. Transcription initiation is a highly regulated step in E. coli, but it has been challenging to study its mechanism because of its stochasticity and complexity. In this review, we describe how single-molecule approaches have contributed to our understanding of transcription and have uncovered mechanistic details that were not observed in conventional assays because of ensemble averaging.

Original languageAmerican English
Pages (from-to)1278-1290
Number of pages13
JournalProtein Science
Volume26
Issue number7
DOIs
StatePublished - 1 Jul 2017
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2017 The Protein Society

Keywords

  • FRET
  • RNA polymerase
  • alternating laser excitation
  • single molecule
  • transcription

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