Multiple binding modes of substrate to the catalytic RNA subunit of RNase P from Escherichia coli

Daniel A. Pomeranz Krummel, Sidney Altman

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

M1 RNA that contained 4'-thiouridine was photochemically cross-linked to different substrates and to a product of the reaction it governs. The locations of the cross-links in these photochemically induced complexes were identified. The cross-links indicated that different substrates share some contacts but have distinct binding modes to M1 RNA. The binding of some substrates also results in a substrate-dependent conformational change in the enzymatic RNA, as evidenced by the appearance of an M1 RNA intramolecular cross-link. The identification of the cross-links between M1 RNA and product indicate that they are shared with only one of the three cross-linked E-S complexes that were identified, an indication of noncompetitive inhibition by the product. We also examined whether the cross-linked complexes between M1 RNA and substrate(s) or product are altered in the presence of the enzyme's protein cofactor (C5 protein) and in the presence of different concentrations of divalent metal ions. C5 protein enhanced the yield of certain M1 RNA- substrate cross-linked complexes for both wild-type M1 RNA and a deletion mutant of M1 RNA (Δ[273-281]), but not for the M1 RNA-product complex. High concentrations of Mg2+ increased the yield of all M1 RNA-substrate complexes but not the M1 RNA-product complex.

Original languageEnglish (US)
Pages (from-to)1021-1033
Number of pages13
JournalRNA
Volume5
Issue number8
DOIs
StatePublished - Aug 1999
Externally publishedYes

Fingerprint

Ribonuclease P
Catalytic RNA
Catalytic Domain
RNA
Escherichia coli
Thiouridine
Proteins
Coenzymes

Keywords

  • E-S complexes
  • M1 RNA
  • Photochemical induced cross-links
  • Substrate-induced fit

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology

Cite this

Multiple binding modes of substrate to the catalytic RNA subunit of RNase P from Escherichia coli. / Pomeranz Krummel, Daniel A.; Altman, Sidney.

In: RNA, Vol. 5, No. 8, 08.1999, p. 1021-1033.

Research output: Contribution to journalArticle

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