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

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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.

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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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Authors and Affiliations

  1. Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117997, Moscow, Russia

    O. O. Zharskaya & O. V. Zatsepina

  2. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, 119992, Moscow, Russia

    A. S. Barsukova & O. V. Zatsepina

Authors
  1. O. O. Zharskaya
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  2. A. S. Barsukova
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  3. O. V. Zatsepina
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Correspondence to O. V. Zatsepina.

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Original Russian Text © O. O. Zharskaya, A. S. Barsukova, O. V. Zatsepina, 2008, published in Biokhimiya, 2008, Vol. 73, No. 4, pp. 504–514.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM07-268, February 24, 2008.

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Zharskaya, O.O., Barsukova, A.S. & Zatsepina, O.V. Effect of roscovitine, a selective cyclin B-dependent kinase 1 inhibitor, on assembly of the nucleolus in mitosis. Biochemistry Moscow 73, 411–419 (2008). https://doi.org/10.1134/S0006297908040056

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  • DOI: https://doi.org/10.1134/S0006297908040056

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