The structure of a transcribing T7 RNA polymerase in transition from initiation to elongation

Kimberly J. Durniak, Scott Bailey, Thomas A. Steitz

Research output: Contribution to journalArticle

Abstract

Structural studies of the T7 bacteriophage DNA-dependent RNA polymerase (T7 RNAP) have shown that the conformation of the amino-terminal domain changes substantially between the initiation and elongation phases of transcription, but how this transition is achieved remains unclear. We report crystal structures of T7 RNAP bound to promoter DNA containing either a 7- or an 8-nucleotide (nt) RNA transcript that illuminate intermediate states along the transition pathway. The amino-terminal domain comprises the C-helix subdomain and the promoter binding domain (PBD), which consists of two segments separated by subdomain H. The structures of the intermediate complex reveal that the PBD and the bound promoter rotate by ∼45° upon synthesis of an 8-nt RNA transcript. This allows the promoter contacts to be maintained while the active site is expanded to accommodate a growing heteroduplex. The C-helix subdomain moves modestly toward its elongation conformation, whereas subdomain H remains in its initiation- rather than its elongation-phase location, more than 70 angstroms away.

Original languageEnglish (US)
Pages (from-to)553-557
Number of pages5
JournalScience
Volume322
Issue number5901
DOIs
StatePublished - Oct 24 2008

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Nucleotides
RNA
Bacteriophage T7
Phase Transition
DNA-Directed RNA Polymerases
Catalytic Domain
DNA
bacteriophage T7 RNA polymerase

ASJC Scopus subject areas

  • General

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The structure of a transcribing T7 RNA polymerase in transition from initiation to elongation. / Durniak, Kimberly J.; Bailey, Scott; Steitz, Thomas A.

In: Science, Vol. 322, No. 5901, 24.10.2008, p. 553-557.

Research output: Contribution to journalArticle

Durniak, Kimberly J. ; Bailey, Scott ; Steitz, Thomas A. / The structure of a transcribing T7 RNA polymerase in transition from initiation to elongation. In: Science. 2008 ; Vol. 322, No. 5901. pp. 553-557.
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