Mitosis-specific Phosphorylation of Gar2, a Fission Yeast Nucleolar Protein Structurally Related to Nucleolin

Overview

Journal Chromosoma

Specialty Molecular Biology

Date 1997 Jun 1

PMID 9211981

Citations 5

Authors

M P Gulli

M Faubladier

H Sicard

M Caizergues-Ferrer

Affiliations

Soon will be listed here.

Abstract

The nucleolar protein gar2 of fission yeast is structurally related to the multifunctional nucleolar protein nucleolin from vertebrates and has been shown to be implicated in production of 18S rRNA. gar2 contains several potential casein kinase 2 (CK2) phosphorylation sites and a single putative p34(cdc2 )phosphorylation site in the consensus S50PKK. Here, we show that, like nucleolin, gar2 is phosphorylated in vitro by both highly purified CK2 from CHO cells and p34(cdc2 )from starfish oocytes. Moreover, the substitution of alanine for the N-terminal serine 50 abolishes phosphorylation by p34(cdc2 )in vitro. We also provide evidence that gar2 is phosphorylated in vitro by a p13(suc1)-Sepharose-bound kinase from Schizosaccharomyces pombe extracts that displays cell cycle-regulated activity similar to that of the p34(cdc2(kinase. In vivo 32P labeling of cells indicates that gar2 is a phosphoprotein and that incorporation of phosphate on residue 50 occurs specifically at mitosis. Taken together, these results lead us to propose that gar2 is likely to be an in vivo substrate for the mitotic p34(cdc2 )kinase. However, this posttranslational modification of the gar2 protein does not appear to be essential for normal production of 18S rRNA.

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References

Roussel P, Hernandez-Verdun D . Identification of Ag-NOR proteins, markers of proliferation related to ribosomal gene activity. Exp Cell Res. 1994; 214(2):465-72. DOI: 10.1006/excr.1994.1283. View

Tyers M, Tokiwa G, Nash R, Futcher B . The Cln3-Cdc28 kinase complex of S. cerevisiae is regulated by proteolysis and phosphorylation. EMBO J. 1992; 11(5):1773-84. PMC: 556635. DOI: 10.1002/j.1460-2075.1992.tb05229.x. View

Weisenberger D, Scheer U . A possible mechanism for the inhibition of ribosomal RNA gene transcription during mitosis. J Cell Biol. 1995; 129(3):561-75. PMC: 2120438. DOI: 10.1083/jcb.129.3.561. View

Pinna L . Casein kinase 2: an 'eminence grise' in cellular regulation?. Biochim Biophys Acta. 1990; 1054(3):267-84. DOI: 10.1016/0167-4889(90)90098-x. View

Gulli M, Girard J, Zabetakis D, Lapeyre B, Melese T, Caizergues-Ferrer M . gar2 is a nucleolar protein from Schizosaccharomyces pombe required for 18S rRNA and 40S ribosomal subunit accumulation. Nucleic Acids Res. 1995; 23(11):1912-8. PMC: 306962. DOI: 10.1093/nar/23.11.1912. View

Gallagher I, Alfa C, Hyams J . p63cdc13, a B-type cyclin, is associated with both the nucleolar and chromatin domains of the fission yeast nucleus. Mol Biol Cell. 1993; 4(11):1087-96. PMC: 275745. DOI: 10.1091/mbc.4.11.1087. View

Lapeyre B, Bourbon H, Amalric F . Nucleolin, the major nucleolar protein of growing eukaryotic cells: an unusual protein structure revealed by the nucleotide sequence. Proc Natl Acad Sci U S A. 1987; 84(6):1472-6. PMC: 304456. DOI: 10.1073/pnas.84.6.1472. View

Kunkel T, Roberts J, Zakour R . Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987; 154:367-82. DOI: 10.1016/0076-6879(87)54085-x. View

Labbe J, Cavadore J, Doree M . M phase-specific cdc2 kinase: preparation from starfish oocytes and properties. Methods Enzymol. 1991; 200:291-301. DOI: 10.1016/0076-6879(91)00147-o. View

10.

McCully E, Robinow C . Mitosis in the fission yeast Schizosaccharomyces pombe: a comparative study with light and electron microscopy. J Cell Sci. 1971; 9(2):475-507. DOI: 10.1242/jcs.9.2.475. View

11.

Nigg E . Cyclin-dependent protein kinases: key regulators of the eukaryotic cell cycle. Bioessays. 1995; 17(6):471-80. DOI: 10.1002/bies.950170603. View

12.

Heald R, McKeon F . Mutations of phosphorylation sites in lamin A that prevent nuclear lamina disassembly in mitosis. Cell. 1990; 61(4):579-89. DOI: 10.1016/0092-8674(90)90470-y. View

13.

Schagger H, von Jagow G . Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987; 166(2):368-79. DOI: 10.1016/0003-2697(87)90587-2. View

14.

Alderuccio F, Chan E, Tan E . Molecular characterization of an autoantigen of PM-Scl in the polymyositis/scleroderma overlap syndrome: a unique and complete human cDNA encoding an apparent 75-kD acidic protein of the nucleolar complex. J Exp Med. 1991; 173(4):941-52. PMC: 2190817. DOI: 10.1084/jem.173.4.941. View

15.

Lee W, Xue Z, Melese T . The NSR1 gene encodes a protein that specifically binds nuclear localization sequences and has two RNA recognition motifs. J Cell Biol. 1991; 113(1):1-12. PMC: 2288927. DOI: 10.1083/jcb.113.1.1. View

16.

Moll T, Tebb G, Surana U, Robitsch H, Nasmyth K . The role of phosphorylation and the CDC28 protein kinase in cell cycle-regulated nuclear import of the S. cerevisiae transcription factor SWI5. Cell. 1991; 66(4):743-58. DOI: 10.1016/0092-8674(91)90118-i. View

17.

Basi G, Schmid E, Maundrell K . TATA box mutations in the Schizosaccharomyces pombe nmt1 promoter affect transcription efficiency but not the transcription start point or thiamine repressibility. Gene. 1993; 123(1):131-6. DOI: 10.1016/0378-1119(93)90552-e. View

18.

Kharrat A, Derancourt J, Doree M, Amalric F, Erard M . Synergistic effect of histone H1 and nucleolin on chromatin condensation in mitosis: role of a phosphorylated heteromer. Biochemistry. 1991; 30(42):10329-36. DOI: 10.1021/bi00106a034. View

19.

Lischwe M, Richards R, BUSCH R, Busch H . Localization of phosphoprotein C23 to nucleolar structures and to the nucleolus organizer regions. Exp Cell Res. 1981; 136(1):101-9. DOI: 10.1016/0014-4827(81)90041-0. View

20.

Xue Z, Melese T . Nucleolar proteins that bind NLSs: a role in nuclear import or ribosome biogenesis?. Trends Cell Biol. 1994; 4(12):414-7. DOI: 10.1016/0962-8924(94)90095-7. View