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PMID: 2388630 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

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

Molecular and cellular biology ·Vol. 10 ·No. 9 ·1990-09-00 ·Pages 4920-31

Kass S, Sollner-Webb B

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 approximately 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.

MeSH Terms
Animals Carcinoma, Ehrlich Tumor/metabolism Cross-Linking Reagents DNA, Ribosomal/genetics Heparin/pharmacology Kinetics Macromolecular Substances Mice Protein Binding RNA Precursors/genetics RNA Processing, Post-Transcriptional/drug effects RNA, Ribosomal/genetics Transcription, Genetic Ultraviolet Rays
Chemicals
Cross-Linking Reagents DNA, Ribosomal Macromolecular Substances RNA Precursors RNA, Ribosomal Heparin
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Kass S
Department of Biological Chemistry, School of Medicine, Johns Hopkins University, Baltimore, Maryland 21205-2185.
Sollner-Webb B
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.10.9.4920-4931.1990
Published
1990-09-00
Pages
4920-31
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC361110
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.10.9.4920-4931.1990 PMC Full Text View journal details
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