Structure of a T7 RNA polymerase elongation complex at 2.9 A resolution

Nature. 2002 Nov 7;420(6911):43-50. doi: 10.1038/nature01129. Epub 2002 Oct 9.

Abstract

The single-subunit bacteriophage T7 RNA polymerase carries out the transcription cycle in an identical manner to that of bacterial and eukaryotic multisubunit enzymes. Here we report the crystal structure of a T7 RNA polymerase elongation complex, which shows that incorporation of an 8-base-pair RNA-DNA hybrid into the active site of the enzyme induces a marked rearrangement of the amino-terminal domain. This rearrangement involves alternative folding of about 130 residues and a marked reorientation (about 130 degrees rotation) of a stable core subdomain, resulting in a structure that provides elements required for stable transcription elongation. A wide opening on the enzyme surface that is probably an RNA exit pathway is formed, and the RNA-DNA hybrid is completely buried in a newly formed, deep protein cavity. Binding of 10 base pairs of downstream DNA is stabilized mostly by long-distance electrostatic interactions. The structure implies plausible mechanisms for the various phases of the transcription cycle, and reveals important structural similarities with the multisubunit RNA polymerases.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Bacteriophage T7 / enzymology*
  • Base Pairing
  • Binding Sites
  • Crystallography, X-Ray
  • DNA / genetics
  • DNA / metabolism
  • DNA-Directed RNA Polymerases / chemistry*
  • DNA-Directed RNA Polymerases / metabolism*
  • Models, Molecular
  • Molecular Sequence Data
  • Promoter Regions, Genetic
  • Protein Binding
  • Protein Folding
  • Protein Structure, Tertiary
  • RNA / biosynthesis
  • RNA / genetics
  • RNA / metabolism
  • Static Electricity
  • Transcription, Genetic
  • Viral Proteins

Substances

  • Viral Proteins
  • RNA
  • DNA
  • bacteriophage T7 RNA polymerase
  • DNA-Directed RNA Polymerases

Associated data

  • PDB/1H38