Structural basis of transcription: Nucleotide selection by rotation in the RNA polymerase II active center

Kenneth D. Westover, David A. Bushnell, Roger D. Kornberg

Research output: Contribution to journalArticle

196 Citations (Scopus)

Abstract

Binding of a ribonucleoside triphosphate to an RNA polymerase II transcribing complex, with base pairing to the template DNA, was revealed by X-ray crystallography. Binding of a mismatched nucleoside triphosphate was also detected, but in an adjacent site, inverted with respect to the correctly paired nucleotide. The results are consistent with a two-step mechanism of nucleotide selection, with initial binding to an entry (E) site beneath the active center in an inverted orientation, followed by rotation into the nucleotide addition (A) site for pairing with the template DNA. This mechanism is unrelated to that of single subunit RNA polymerases and so defines a new paradigm for the large, multisubunit enzymes. Additional findings from these studies include a third nucleotide binding site that may define the length of backtracked RNA; DNA double helix unwinding in advance of the polymerase active center; and extension of the diffraction limit of RNA polymerase II crystals to 2.3 Å.

Original languageEnglish (US)
Pages (from-to)481-489
Number of pages9
JournalCell
Volume119
Issue number4
DOIs
StatePublished - Nov 12 2004

Fingerprint

RNA Polymerase II
Transcription
Nucleotides
DNA
Ribonucleosides
X ray crystallography
X Ray Crystallography
DNA-Directed RNA Polymerases
Nucleosides
Base Pairing
Catalytic Domain
Diffraction
Binding Sites
RNA
Crystals
Enzymes
triphosphoric acid

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

Structural basis of transcription : Nucleotide selection by rotation in the RNA polymerase II active center. / Westover, Kenneth D.; Bushnell, David A.; Kornberg, Roger D.

In: Cell, Vol. 119, No. 4, 12.11.2004, p. 481-489.

Research output: Contribution to journalArticle

Westover, Kenneth D. ; Bushnell, David A. ; Kornberg, Roger D. / Structural basis of transcription : Nucleotide selection by rotation in the RNA polymerase II active center. In: Cell. 2004 ; Vol. 119, No. 4. pp. 481-489.
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