Histone-specific RNA 3′ processing in nuclear extracts from mammalian cells

Claudia Stauber, Dominique Soldati, Bernhard Luscher, Daniel Schümperli

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

The mature 3' ends of histone mRNAs are formed by endonucleolytic cleavage of longer precursor transcripts. This process occurs in the nucleus and can be regarded as the equivalent of the polyadenylation reaction involved in 3′-end-generation of all other mRNAs. A sea urchin H3 gene that failed to be properly processed in the Xenopus oocyte system proved particularly useful, because it allowed the identification of a processing component from sea urchins by a complementation assay. Nuclear extracts prepared from cells under various growth conditions have helped to reveal proliferation-dependent changes in the efficiency of histone RNA 3′ processing. RNA substrates for in vitro processing are best prepared by runoff transcription of specific DNA templates with bacterial or phage RNA polymerases. For this purpose, a restriction fragment containing the 3′-terminal region of a histone gene and including the conserved palindrome and spacer motifs is cloned into a polylinker sequence downstream of a strong promoter.

Original languageEnglish (US)
Pages (from-to)74-89
Number of pages16
JournalMethods in enzymology
Volume181
Issue numberC
DOIs
StatePublished - Jan 1 1990

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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