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Published online 7 August 2000. doi:10.1083/jcb.150.3.433
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© The Rockefeller University Press, 0021-9525/2000//433 $5.00
The Journal of Cell Biology, Volume 150, Number 3, , 2000 433-446

Original Article

The Dynamics of Postmitotic Reassembly of the Nucleolus



Miroslav Dundra,b, Tom Mistelib, and Mark O.J. Olsona

a Department of Biochemistry, University of Mississippi Medical Center, Jackson Mississippi 39216
b National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892
2500 North State St., Jackson, MS 39216-4505.(601) 984-1501(601) 984-1500

molson{at}biochem.umsmed.edu

Mammalian cell nucleoli disassemble at the onset of M-phase and reassemble during telophase. Recent studies showed that partially processed preribosomal RNA (pre-rRNA) is preserved in association with processing components in the perichromosomal regions (PRs) and in particles called nucleolus-derived foci (NDF) during mitosis. Here, the dynamics of nucleolar reassembly were examined for the first time in living cells expressing fusions of the processing-related proteins fibrillarin, nucleolin, or B23 with green fluorescent protein (GFP). During telophase the NDF disappeared with a concomitant appearance of material in the reforming nuclei. Prenucleolar bodies (PNBs) appeared in nuclei in early telophase and gradually disappeared as nucleoli formed, strongly suggesting the transfer of PNB components to newly forming nucleoli. Fluorescence recovery after photobleaching (FRAP) showed that fibrillarin-GFP reassociates with the NDF and PNBs at rapid and similar rates. The reentry of processing complexes into telophase nuclei is suggested by the presence of pre-rRNA sequences in PNBs. Entry of specific proteins into the nucleolus approximately correlated with the timing of processing events. The mitotically preserved processing complexes may be essential for regulating the distribution of components to reassembling daughter cell nucleoli.

Key Words: mitosis • nucleolus • prenucleolar body • pre-rRNA processing • rDNA transcription

© 2000 The Rockefeller University Press

The online version of this article contains supplemental material.

Abbreviations used in this paper: DFC, dense fibrillar component; ETS, external transcribed spacer; FC, fibrillar center; NDF, nucleolus-derived foci; NOR, nucleolus organizer region; PNB, prenucleolar body; PR, perichromosomal region; RNA pol I, RNA polymerase I; UBF, upstream binding factor.


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