Spatial organization of transcription elongation complex in Escherichia coli

Science. 1998 Jul 17;281(5375):424-8. doi: 10.1126/science.281.5375.424.

Abstract

During RNA synthesis in the ternary elongation complex, RNA polymerase enzyme holds nucleic acids in three contiguous sites: the double-stranded DNA-binding site (DBS) ahead of the transcription bubble, the RNA-DNA heteroduplex-binding site (HBS), and the RNA-binding site (RBS) upstream of HBS. Photochemical cross-linking allowed mapping of the DNA and RNA contacts to specific positions on the amino acid sequence. Unexpectedly, the same protein regions were found to participate in both DBS and RBS. Thus, DNA entry and RNA exit occur close together in the RNA polymerase molecule, suggesting that the three sites constitute a single unit. The results explain how RNA in the integrated unit RBS-HBS-DBS may stabilize the ternary complex, whereas a hairpin in RNA result in its dissociation.

Publication types

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

MeSH terms

  • Binding Sites
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / metabolism*
  • DNA-Directed RNA Polymerases / chemistry
  • DNA-Directed RNA Polymerases / metabolism*
  • Escherichia coli / genetics*
  • Escherichia coli / metabolism
  • Idoxuridine / metabolism
  • Models, Genetic
  • Nucleic Acid Conformation
  • Nucleic Acid Heteroduplexes / metabolism*
  • Protein Binding
  • RNA, Bacterial / chemistry
  • RNA, Bacterial / metabolism*
  • Templates, Genetic
  • Transcription, Genetic*
  • Ultraviolet Rays

Substances

  • DNA, Bacterial
  • Nucleic Acid Heteroduplexes
  • RNA, Bacterial
  • DNA-Directed RNA Polymerases
  • Idoxuridine