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Biochemistry (Moscow)

, 73:411 | Cite as

Effect of roscovitine, a selective cyclin B-dependent kinase 1 inhibitor, on assembly of the nucleolus in mitosis

  • O. O. Zharskaya
  • A. S. Barsukova
  • O. V. ZatsepinaEmail author
Article

Abstract

It is well known that at the beginning of mitosis the nucleolus disassembles but then reassembles at the end of mitosis. However, the mechanisms of these processes are still unclear. In the present work, we show for the first time that selective inhibition of cyclin B-dependent kinase 1 (CDK1) by roscovitine induces premature assembly of the nucleolus in mammalian cells in metaphase. Treatment of metaphase cells with roscovitine induces formation of structures in their cytoplasm that contain major proteins of the mature nucleolus participating in rRNA processing, such as B23/nucleophosmin, C23/nucleolin, fibrillarin, Nop52, as well as partially processed (immature) 46–45S pre-rRNA. This effect is reproducible in cells of various types; this indicates that general mechanisms regulate early stages of the nucleolus reassembly with CDK1 participation in mammalian cells. Based on our and literature data, we suggest that inactivation of the CDK1-cyclin B complex at the end of mitosis results in dephosphorylation of B23/nucleophosmin and C23/nucleolin; this facilitates their interaction with pre-rRNA and leads to formation of insoluble supramolecular complexes-nucleolus-derived foci.

Key words

nucleolus mitosis cyclin B-dependent kinase CDK1 nucleolus-derived foci (NDF) fibrillarin B23/nucleophosmin roscovitine 

Abbreviations

BrUTP

5-bromouridine-5′-triphosphate

CDK1

cyclin B-dependent kinase 1

DAPI

4′,6-diamidino-2-phenylindole

5′ETS

5′-external transcribed spacer

FISH

fluorescent in situ hybridization

FITC

fluorescein isothiocyanate

iNDF

induced NDF

NDF

nucleolus-derived focus

NOR

nucleolar organizing region

PBS

phosphate buffered saline

pre-rRNA

47-45S ribosomal RNA precursor

rDNA

ribosomal DNA

rRNA

ribosomal RNA

SSC

standard saline citrate

UBF

upstream binding factor

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Copyright information

© MAIK Nauka 2008

Authors and Affiliations

  • O. O. Zharskaya
    • 1
  • A. S. Barsukova
    • 2
  • O. V. Zatsepina
    • 1
    • 2
    Email author
  1. 1.Shemyakin and Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussia
  2. 2.Belozersky Institute of Physico-Chemical BiologyLomonosov Moscow State UniversityMoscowRussia

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