Identification of a ribonuclease P-like activity from human KB cells

Raymond A. Koski, Alfred L M Bothwell, Sidney Altman

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

An endoribonuclease which cleaves tRNA precursor molecules has been partially purified from human KB tissue culture cells. This activity is found in cytoplasmic fractions but is not detectable in the nucleoplasm. tRNA precursor molecules from both E. coli and KB cells are cleaved by this novel activity to produce 5′ phosphate-terminated oligonucleotides. E. coli RNAase P and the KB cell nuclease both make a single endonucleolytic scission in E. coli tRNATyr precursor, thereby separating the 41 extra nucleotides on the 5′ end of the precursor molecule from the 5′ terminal sequence of the mature tRNATyr molecule. The cleavage products generated from other E. coli tRNA precursors by the KB cell activity are identical in size to those produced by RNAase P. The KB cell endoribonuclease requires Mg2- and a monovalent cation (Na+, K+, or NH4 +) for function. The enzymatic activity has a broad pH optimum, centered near pH 8.0, and the activity is inhibited by tRNA. Several KB cell RNAs with long half-lives in vivo, including 5S and bulk 4S RNA, are not cleaved by this nuclease. The KB cell endoribonuclease resembles E. coli RNAase P in its substrate specificity, pH optimum, ion requirements, and sensitivity to tRNA. These properties and the cytoplasmic localization of the novel endoribonuclease indicate its involvement in the biosynthesis of KB cell tRNA.

Original languageEnglish (US)
Pages (from-to)101-116
Number of pages16
JournalCell
Volume9
Issue number1
DOIs
StatePublished - 1976
Externally publishedYes

Fingerprint

Ribonuclease P
KB Cells
Endoribonucleases
Human Activities
Escherichia coli
RNA Precursors
RNA, Transfer, Tyr
Transfer RNA
Molecules
RNA
Monovalent Cations
Tissue culture
Biosynthesis
Oligonucleotides
Nucleotides
Phosphates
Substrate Specificity
Ions
Cell Culture Techniques
Substrates

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Identification of a ribonuclease P-like activity from human KB cells. / Koski, Raymond A.; Bothwell, Alfred L M; Altman, Sidney.

In: Cell, Vol. 9, No. 1, 1976, p. 101-116.

Research output: Contribution to journalArticle

Koski, Raymond A. ; Bothwell, Alfred L M ; Altman, Sidney. / Identification of a ribonuclease P-like activity from human KB cells. In: Cell. 1976 ; Vol. 9, No. 1. pp. 101-116.
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