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PMID: 10791972 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Association of yeast RNA polymerase I with a nucleolar substructure active in rRNA synthesis and processing.

The Journal of cell biology ·Vol. 149 ·No. 3 ·2000-05-01 ·Pages 575-90

Fath S, Milkereit P, Podtelejnikov AV, Bischler N, Schultz P, Bier M, Mann M, Tschochner H

Abstract

A novel ribonucleoprotein complex enriched in nucleolar proteins was purified from yeast extracts and constituents were identified by mass spectrometry. When isolated from rapidly growing cells, the assembly contained ribonucleic acid (RNA) polymerase (pol) I, and some of its transcription factors like TATA-binding protein (TBP), Rrn3p, Rrn5p, Rrn7p, and Reb1p along with rRNA processing factors, like Nop1p, Cbf5p, Nhp2p, and Rrp5p. The small nucleolar RNAs (snoRNAs) U3, U14, and MRP were also found to be associated with the complex, which supports accurate transcription, termination, and pseudouridylation of rRNA. Formation of the complex did not depend on pol I, and the complex could efficiently recruit exogenous pol I into active ribosomal DNA (rDNA) transcription units. Visualization of the complex by electron microscopy and immunogold labeling revealed a characteristic cluster-forming network of nonuniform size containing nucleolar proteins like Nop1p and Fpr3p and attached pol I. Our results support the idea that a functional nucleolar subdomain formed independently of the state of rDNA transcription may serve as a scaffold for coordinated rRNA synthesis and processing.

MeSH Terms
Cell Nucleolus/metabolism DNA, Ribosomal/genetics Macromolecular Substances Mass Spectrometry Microscopy, Immunoelectron Nuclear Proteins/chemistry RNA Polymerase I/genetics,metabolism RNA, Ribosomal/biosynthesis,metabolism RNA, Small Nucleolar/chemistry Ribonucleoproteins/chemistry Transcription Factors/chemistry Transcription, Genetic Yeasts
Chemicals
DNA, Ribosomal Macromolecular Substances Nuclear Proteins RNA, Ribosomal RNA, Small Nucleolar Ribonucleoproteins Transcription Factors RNA Polymerase I
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Fath S
Biochemie-Zentrum Heidelberg, D-69120 Heidelberg, Germany.
Milkereit P
Podtelejnikov A V
Bischler N
Schultz P
Bier M
Mann M
Tschochner H
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.149.3.575
Published
2000-05-01
Pages
575-90
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2174860
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.149.3.575 PMC Full Text View journal details
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