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PMID: 9880328 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Association of nonribosomal nucleolar proteins in ribonucleoprotein complexes during interphase and mitosis.

Molecular biology of the cell ·Vol. 10 ·No. 1 ·1999-01-00 ·Pages 77-90

Piñol-Roma S

Abstract

rRNA precursors are bound throughout their length by specific proteins, as the pre-rRNAs emerge from the transcription machinery. The association of pre-rRNA with proteins as ribonucleoprotein (RNP) complexes persists during maturation of 18S, 5.8S, and 28S rRNA, and through assembly of ribosomal subunits in the nucleolus. Preribosomal RNP complexes contain, in addition to ribosomal proteins, an unknown number of nonribosomal nucleolar proteins, as well as small nucleolar RNA-ribonucleoproteins (sno-RNPs). This report describes the use of a specific, rapid, and mild immunopurification approach to isolate and analyze human RNP complexes that contain nonribosomal nucleolar proteins, as well as ribosomal proteins and rRNA. Complexes immunopurified with antibodies to nucleolin-a major nucleolar RNA-binding protein-contain several distinct specific polypeptides that include, in addition to nucleolin, the previously identified nucleolar proteins B23 and fibrillarin, proteins with electrophoretic mobilities characteristic of ribosomal proteins including ribosomal protein S6, and a number of additional unidentified proteins. The physical association of these proteins with one another is mediated largely by RNA, in that the complexes dissociate upon digestion with RNase. Complexes isolated from M-phase cells are similar in protein composition to those isolated from interphase cell nuclear extracts. Therefore, the predominant proteins that associate with nucleolin in interphase remain in RNP complexes during mitosis, despite the cessation of rRNA synthesis and processing in M-phase. In addition, precursor rRNA, as well as processed 18S and 28S rRNA and candidate rRNA processing intermediates, is found associated with the immunopurified complexes. The characteristics of the rRNP complexes described here, therefore, indicate that they represent bona fide precursors of mature cytoplasmic ribosomal subunits.

MeSH Terms
Animals Base Sequence Chromosomal Proteins, Non-Histone/isolation & purification,metabolism DNA Probes/genetics HeLa Cells Humans Immunohistochemistry Interphase/physiology Macromolecular Substances Mice Mitosis/physiology Nuclear Proteins/isolation & purification,metabolism Nucleophosmin Phosphoproteins/isolation & purification,metabolism RNA Precursors/genetics,isolation & purification,metabolism RNA-Binding Proteins/isolation & purification,metabolism Ribonucleoproteins/isolation & purification,metabolism Ribosomal Protein S6 Ribosomal Proteins/isolation & purification,metabolism Ribosomes/chemistry,metabolism
Chemicals
Chromosomal Proteins, Non-Histone DNA Probes Macromolecular Substances Npm1 protein, mouse Nuclear Proteins Phosphoproteins RNA Precursors RNA-Binding Proteins Ribonucleoproteins Ribosomal Protein S6 Ribosomal Proteins fibrillarin nucleolin Nucleophosmin
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Piñol-Roma S
Department of Cell Biology and Anatomy, Mount Sinai School of Medicine, New York, New York 10029-6574, USA.
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.10.1.77
Published
1999-01-00
Pages
77-90
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC25155
Subset
IM
External Links
PubMed source DOI: 10.1091/mbc.10.1.77 PMC Full Text View journal details
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