Localisation of the binding site for the initiating substrate on the RNA polymerase from Sulfolobus acidocaldarius

Mikhail A. Grachev, Arkadij A. Mustaev, Evgeny F. Zaychikov, Anton J. Lindner, Guido R. Hartmann

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

RNA polymerase from the archaebacterium Sulfolobus acidocaldarius was chemically modified with AMP o-formylphenyl ester followed by reduction with borohydride. The modified protein catalyzes the labeling of its own largest subunit when incubated with [α-33P]UTP in the presence of poly[d(A-T)]. On cleaving of the labeled protein using cyanogen bromide, hydroxylamine or amino acid-specific endoproteinases for a very brief period, the pattern and size of the radioactive fragments formed are best explained by attachment of the label between Gly843 and Met895 of the largest subunit. In this region there exists a highly conserved sequence which is also found in other archaebacterial, eukaryotic and prokaryotic RNA polymerases. This suggests that the binding site for the initiating substrate of RNA polymerases has been conserved during evolution.

Original languageEnglish (US)
Pages (from-to)317-322
Number of pages6
JournalFEBS Letters
Volume250
Issue number2
DOIs
StatePublished - Jul 3 1989

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Keywords

  • (Sulfolobus acidocaldarius)
  • Active center
  • Affinity labeling
  • Localization
  • RNA polymerase

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