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PMID: 1522108 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

In vitro assembly of prenucleolar bodies in Xenopus egg extract.

The Journal of cell biology ·Vol. 118 ·No. 6 ·1992-09-00 ·Pages 1297-304

Bell P, Dabauvalle MC, Scheer U

Abstract

Nuclei assembled in Xenopus egg extract from purified DNA or chromatin resemble their natural counterparts in a number of structural and functional features. However, the most obvious structural element of normal interphase nuclei, the nucleolus, is absent from the in vitro reconstituted nuclei. By EM, cytological silver staining, and immunofluorescence microscopy employing antibodies directed against various nucleolar components we show that nuclei assembled in vitro contain numerous distinct aggregates that resemble prenucleolar bodies (PNBs) by several criteria. Formation of these PNB-like structures requires pore complex-mediated nuclear transport of proteins but is independent of the genetic content of the in vitro nuclei as well as transcriptional and translational events. Our data indicate that nuclei assembled in vitro are capable of initiating early steps of nucleologenesis but that the resulting PNBs are unable to fuse with each other, probably due to the absence of a functional nucleolus organizer. With appropriate modifications, this experimental system should be useful to define and analyze conditions promoting the site-specific assembly of PNBs into a coherent nucleolar body.

MeSH Terms
Animals Cell Extracts Cell Nucleolus/metabolism,ultrastructure Cell Nucleus/metabolism,ultrastructure Fluorescent Antibody Technique Male Microscopy, Electron Nucleic Acid Hybridization Oocytes/metabolism Silver Staining Spermatozoa/metabolism Xenopus/metabolism
Chemicals
Cell Extracts
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Bell P
Institute of Zoology I, Biocenter of the University of Würzburg, Germany.
Dabauvalle M C
Scheer U
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.118.6.1297
Published
1992-09-00
Pages
1297-304
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2289602
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.118.6.1297 PMC Full Text View journal details
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