The structural mechanism of translocation and helicase activity in T7 RNA polymerase

Yuhui Yin, Thomas A. Steitz

Research output: Contribution to journalArticle

229 Citations (Scopus)

Abstract

RNA polymerase functions like a molecular motor that can convert chemical energy into the work of strand separation and translocation along the DNA during transcription. The structures of phage T7 RNA polymerase in an elongation phase substrate complex that includes the incoming nucleoside triphosphate and a pretranslocation product complex that includes the product pyrophosphate (PPi) are described here. These structures and the previously determined posttranslocation elongation complex demonstrate that two enzyme conformations exist during a cycle of single nucleotide addition. One orientation of a five-helix subdomain is stabilized by the phosphates of either the incoming NTP or by the product PPi. A second orientation of this subdomain is stable in their absence and is associated with translocation of the heteroduplex product as well as strand separation of the downstream DNA. We propose that the dissociation of the product PPi after nucleotide addition produces the protein conformational change resulting in translocation and strand separation.

Original languageEnglish (US)
Pages (from-to)393-404
Number of pages12
JournalCell
Volume116
Issue number3
DOIs
StatePublished - Feb 6 2004
Externally publishedYes

Fingerprint

Nucleotides
Bacteriophage T7
Elongation
DNA
DNA-Directed RNA Polymerases
Nucleosides
Bacteriophages
Phosphates
Transcription
Conformations
Enzymes
Proteins
Substrates
bacteriophage T7 RNA polymerase
diphosphoric acid
triphosphoric acid

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

The structural mechanism of translocation and helicase activity in T7 RNA polymerase. / Yin, Yuhui; Steitz, Thomas A.

In: Cell, Vol. 116, No. 3, 06.02.2004, p. 393-404.

Research output: Contribution to journalArticle

Yin, Yuhui ; Steitz, Thomas A. / The structural mechanism of translocation and helicase activity in T7 RNA polymerase. In: Cell. 2004 ; Vol. 116, No. 3. pp. 393-404.
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