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PMID: 11381093 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Nucleolar assembly of the rRNA processing machinery in living cells.

The Journal of cell biology ·Vol. 153 ·No. 5 ·2001-05-28 ·Pages 1097-110

Savino TM, Gébrane-Younès J, De Mey J, Sibarita JB, Hernandez-Verdun D

Abstract

To understand how nuclear machineries are targeted to accurate locations during nuclear assembly, we investigated the pathway of the ribosomal RNA (rRNA) processing machinery towards ribosomal genes (nucleolar organizer regions [NORs]) at exit of mitosis. To follow in living cells two permanently transfected green fluorescence protein-tagged nucleolar proteins, fibrillarin and Nop52, from metaphase to G1, 4-D time-lapse microscopy was used. In early telophase, fibrillarin is concentrated simultaneously in prenucleolar bodies (PNBs) and NORs, whereas PNB-containing Nop52 forms later. These distinct PNBs assemble at the chromosome surface. Analysis of PNB movement does not reveal the migration of PNBs towards the nucleolus, but rather a directional flow between PNBs and between PNBs and the nucleolus, ensuring progressive delivery of proteins into nucleoli. This delivery appeared organized in morphologically distinct structures visible by electron microscopy, suggesting transfer of large complexes. We propose that the temporal order of PNB assembly and disassembly controls nucleolar delivery of these proteins, and that accumulation of processing complexes in the nucleolus is driven by pre-rRNA concentration. Initial nucleolar formation around competent NORs appears to be followed by regroupment of the NORs into a single nucleolus 1 h later to complete the nucleolar assembly. This demonstrates the formation of one functional domain by cooperative interactions between different chromosome territories.

MeSH Terms
Blotting, Western Cell Nucleolus/chemistry,genetics,metabolism,ultrastructure Chromosomal Proteins, Non-Histone/metabolism Fluorescent Antibody Technique G1 Phase Genes, rRNA/genetics HeLa Cells Humans Kinetics Microscopy, Electron Mitosis Nuclear Proteins/genetics,metabolism Nucleolus Organizer Region/genetics,metabolism,ultrastructure Phosphoproteins/metabolism Protein Transport Protozoan Proteins RNA Polymerase I/metabolism RNA Processing, Post-Transcriptional RNA, Ribosomal/genetics,metabolism Recombinant Fusion Proteins/metabolism Telophase Transfection
Chemicals
Chromosomal Proteins, Non-Histone NOPP52 protein, Tetrahymena thermophila Nuclear Proteins Phosphoproteins Protozoan Proteins RNA, Ribosomal Recombinant Fusion Proteins fibrillarin RNA Polymerase I
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Savino T M
Institut Jacques Monod, UMR 7592, 2 Place Jussieu, 75251 Paris Cedex 05, France.
Gébrane-Younès J
De Mey J
Sibarita J B
Hernandez-Verdun D
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.153.5.1097
Published
2001-05-28
Pages
1097-110
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2174343
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.153.5.1097 PMC Full Text View journal details
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