The first pre-rRNA-processing event occurs in a large complex: Analysis by gel retardation, sedimentation, and UV cross-linking

Susan Kass, Barbara Sollner-Webb

Research output: Contribution to journalArticle

Abstract

The first processing event that mouse pre-rRNA undergoes occurs within the external transcribed spacer and is efficiently reproduced in vitro. Analysis with nondenaturing polyacrylamide gels revealed the formation of heparin-resistant complexes of retarded electrophoretic mobility on the substrate rRNA. The specificity of these complexes was demonstrated by their elimination due to competition with processing-competent, but not with processing-incompetent, rRNAs. Furthermore, complex formation, like the processing cleavage, required only 28 nucleotides of rRNA sequence adjacent to the processing site but was stimulated by additional downstream conserved sequences. These processing complexes formed in a time-dependent manner, and once assembled, they were stable to challenge by competitor rRNA and remained on the processed rRNA. Their sedimentation coefficient was ∼20S. UV cross-linking studies with 4-thiouridine-substituted rRNA have identified six polypeptides, 52 to 250 kilodaltons, that are specifically bound to the rRNA processing substrate.

Original languageEnglish (US)
Pages (from-to)4920-4931
Number of pages12
JournalMolecular and cellular biology
Volume10
Issue number9
DOIs
StatePublished - Sep 1990

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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