Evidence for helical unwinding of an RNA substrate by the RNA enzyme RNase P: Use of an interstrand disulfide crosslink in substrate

Daniel A Pomeranz Krummel, Oliver Kent, Andrew M. MacMillan, Sidney Altman

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

To gain an understanding of structural changes induced in substrates by Escherichia coli ribonuclease P (RNase P), we have incorporated an interstrand disulfide crosslink proximal to the cleavage site in a model substrate. RNase P is able to process the reduced, non-crosslinked form of this substrate as well as a substrate in which the free thiol molecules have been alkylated with iodoacetamide. However, the oxidized, cross-linked form is cleaved at a significantly lower rate. Therefore, helical unwinding of the analog of the aminoacyl stem of the substrate near its site of cleavage may be necessary for efficient processing by E. coli RNase P. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)1113-1118
Number of pages6
JournalJournal of Molecular Biology
Volume295
Issue number5
DOIs
StatePublished - Feb 4 2000
Externally publishedYes

Fingerprint

Ribonuclease P
Disulfides
RNA
Enzymes
Escherichia coli
Iodoacetamide
Sulfhydryl Compounds

Keywords

  • Convertible nucleosides
  • Disulfide crosslink
  • M1 RNA
  • Model substrates
  • RNase P

ASJC Scopus subject areas

  • Virology

Cite this

Evidence for helical unwinding of an RNA substrate by the RNA enzyme RNase P : Use of an interstrand disulfide crosslink in substrate. / Krummel, Daniel A Pomeranz; Kent, Oliver; MacMillan, Andrew M.; Altman, Sidney.

In: Journal of Molecular Biology, Vol. 295, No. 5, 04.02.2000, p. 1113-1118.

Research output: Contribution to journalArticle

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