Substrate recognition by human RNase P: Identification of small, model substrates for the enzyme

Y. Yuan, Sidney Altman

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

RNase P from HeLa cells can efficiently cleave tRNA precursor molecules in vitro but cannot cleave potential substrates from which the D, anticodon and variable loops and stems of the tRNA moiety have all been removed. However, molecules from which the latter subdomains have been removed individually do serve as substrates. We show here that molecules that contain only a 5' leader sequence, the acceptor stem and the T stem and loop of the tRNA domain, and a bulge as small as one nucleotide downstream from nucleotide 7 in the tRNA sequence at the junction of the two stems, can serve as substrates for human RNase P. The identity of the nucleotide in the bulge is important in determining both the efficiency of the cleavage and the conformation of the substrate and/or the enzyme-substrate complex. We also show that the human enzyme locates the appropriate site for cleavage of its substrates in part by 'measuring' the length of the helices in the acceptor and T stems in both model and natural substrates.

Original languageEnglish (US)
Pages (from-to)159-168
Number of pages10
JournalEMBO Journal
Volume14
Issue number1
StatePublished - 1995
Externally publishedYes

Fingerprint

Ribonuclease P
Transfer RNA
Identification (control systems)
Nucleotides
Substrates
Enzymes
Anticodon
RNA Precursors
HeLa Cells
Molecules
Conformations

Keywords

  • Bulged nucleotide
  • Human RNase P
  • Measuring RNA
  • Model substrates

ASJC Scopus subject areas

  • Cell Biology
  • Genetics

Cite this

Substrate recognition by human RNase P : Identification of small, model substrates for the enzyme. / Yuan, Y.; Altman, Sidney.

In: EMBO Journal, Vol. 14, No. 1, 1995, p. 159-168.

Research output: Contribution to journalArticle

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