[en] A novel gene, EFHC1, mutated in juvenile myoclonic epilepsy (JME) encodes a protein with three DM10 domains of unknown function and one putative EF-hand motif. To study the properties of EFHC1, we expressed EGFP-tagged protein in various cell lines. In interphase cells, the fusion protein was present in the cytoplasm and in the nucleus with specific accumulation at the centrosome. During mitosis EGFP-EFHC1 colocalized with the mitotic spindle, especially at spindle poles and with the midbody during cytokinesis. Using a specific antibody, we demonstrated the same distribution of the endogenous protein. Deletion analyses revealed that the N-terminal region of EFHC1 is crucial for the association with the mitotic spindle and the midbody. Our results suggest that EFHC1 could play an important role during cell division. (c) 2006 Elsevier Inc. All rights reserved.
Disciplines :
Oncology
Author, co-author :
de Nijs, Laurence ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Biochimie et physiologie humaine et pathologique
Lakaye, Bernard ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Biochimie et physiologie humaine et pathologique
Coumans, Bernard ; Université de Liège - ULiège > CNCM/ Centre fac. de rech. en neurobiologie cell. et moléc.
Leon, Christine ; Université de Liège - ULiège > CNCM/ Centre fac. de rech. en neurobiologie cell. et moléc.
Ikeda, T.
Delgado-Escueta, A. V.
Grisar, Thierry ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Biochimie et physiologie humaine et pathologique
Chanas, G.
Language :
English
Title :
EFHC1, a protein mutated in juvenile myoclonic epilepsy, associates with the mitotic spindle through its N-terminus
Publication date :
10 September 2006
Journal title :
Experimental Cell Research
ISSN :
0014-4827
eISSN :
1090-2422
Publisher :
Elsevier Inc, San Diego, United States - California
Suzuki T., Delgado-Escueta A.V., Aguan K., Alonso M.E., Shi J., Hara Y., Nishida M., Numata T., Medina M.T., Takeuchi T., Morita R., Bai D., Ganesh S., Sugimoto Y., Inazawa J., Bailey J.N., Ochoa A., Jara-Prado A., Rasmussen A., Ramos-Peek J., Cordova S., Rubio-Donnadieu F., Inoue Y., Osawa M., Kaneko S., Oguni H., Mori Y., and Yamakawa K. Mutations in EFHC1 cause juvenile myoclonic epilepsy. Nat. Genet. 36 (2004) 842-849
Braunewell K.H., and Gundelfinger E.D. Intracellular neuronal calcium sensor proteins: a family of EF-hand calcium-binding proteins in search of a function. Cell Tissue Res. 295 (1999) 1-12
Mori Y., Hara Y., Aguan K., Katano M., Yamamoto S., Suzuki T., and Yamakawa K. Molecular and functional interaction of the juvenile myoclonic epilepsy-related gene EFHC1 with the redox-sensitive TRPM2 channel (2005), Society for Neuroscience, Washington
Ikeda T., Ikeda K., Enomoto M., Park M.K., Hirono M., and Kamiya R. The mouse ortholog of EFHC1 implicated in juvenile myoclonic epilepsy is an axonemal protein widely conserved among organisms with motile cilia and flagella. FEBS Lett. 579 (2005) 819-822
Ikeda K., Brown J.A., Yagi T., Norrander J.M., Hirono M., Eccleston E., Kamiya R., and Linck R.W. Rib72, a conserved protein associated with the ribbon compartment of flagellar A-microtubules and potentially involved in the linkage between outer doublet microtubules. J. Biol. Chem. 278 (2003) 7725-7734
Patel-King R.S., Benashski S.E., and King S.M. A bipartite Ca2+-regulated nucleoside-diphosphate kinase system within the Chlamydomonas flagellum. The regulatory subunit p72. J. Biol. Chem. 277 (2002) 34271-34279
Ogawa H., Inouye S., Tsuji F.I., Yasuda K., and Umesono K. Localization, trafficking, and temperature-dependence of the Aequorea green fluorescent protein in cultured vertebrate cells. Proc. Natl. Acad. Sci. U. S. A. 92 (1995) 11899-11903
Oakley B.R. gamma-Tubulin. Curr. Top Dev. Biol. 49 (2000) 27-54
Mastronarde D.N., McDonald K.L., Ding R., and McIntosh J.R. Interpolar spindle microtubules in PTK cells. J. Cell Biol. 123 (1993) 1475-1489
Sharp D.J., McDonald K.L., Brown H.M., Matthies H.J., Walczak C., Vale R.D., Mitchison T.J., and Scholey J.M. The bipolar kinesin, KLP61F, cross-links microtubules within interpolar microtubule bundles of Drosophila embryonic mitotic spindles. J. Cell Biol. 144 (1999) 125-138
Zeitlin S.G., and Sullivan K.F. Animal cytokinesis: breaking up is hard to do. Curr. Biol. 11 (2001) R514-R516
Skop A.R., Liu H., Yates III J., Meyer B.J., and Heald R. Dissection of the mammalian midbody proteome reveals conserved cytokinesis mechanisms. Science 305 (2004) 61-66
Kato T., Watanabe N., Morishima Y., Fujita A., Ishizaki T., and Narumiya S. Localization of a mammalian homolog of diaphanous, mDia1, to the mitotic spindle in HeLa cells. J. Cell Sci. 114 (2001) 775-784
Yan X., Li F., Liang Y., Shen Y., Zhao X., Huang Q., and Zhu X. Human Nudel and NudE as regulators of cytoplasmic dynein in poleward protein transport along the mitotic spindle. Mol. Cell. Biol. 23 (2003) 1239-1250
Keller L.C., Romijn E.P., Zamora I., Yates III J.R., and Marshall W.F. Proteomic analysis of isolated Chlamydomonas centrioles reveals orthologs of ciliary-disease genes. Curr. Biol. 15 (2005) 1090-1098
Gartner W., Rossbacher J., Zierhut B., Daneva T., Base W., Weissel M., Waldhausl W., Pasternack M.S., and Wagner L. The ATP-dependent helicase RUVBL1/TIP49a associates with tubulin during mitosis. Cell Motil. Cytoskeleton 56 (2003) 79-93
Stenoien D.L., Sen S., Mancini M.A., and Brinkley B.R. Dynamic association of a tumor amplified kinase, Aurora-A, with the centrosome and mitotic spindle. Cell Motil. Cytoskeleton 55 (2003) 134-146
Cassimeris L., and Morabito J. TOGp, the human homolog of XMAP215/Dis1, is required for centrosome integrity, spindle pole organization, and bipolar spindle assembly. Mol. Biol. Cell 15 (2004) 1580-1590
Patzke S., Hauge H., Sioud M., Finne E.F., Sivertsen E.A., Delabie J., Stokke T., and Aasheim H.C. Identification of a novel centrosome/microtubule-associated coiled-coil protein involved in cell-cycle progression and spindle organization. Oncogene 24 (2005) 1159-1173
Steffen W., Fajer E.A., and Linck R.W. Centrosomal components immunologically related to tektins from ciliary and flagellar microtubules. J. Cell Sci. 107 Pt. 8 (1994) 2095-2105
Li K., Xu E.Y., Cecil J.K., Turner F.R., Megraw T.L., and Kaufman T.C. Drosophila centrosomin protein is required for male meiosis and assembly of the flagellar axoneme. J. Cell Biol. 141 (1998) 455-467
Larsson M., Norrander J., Graslund S., Brundell E., Linck R., Stahl S., and Hoog C. The spatial and temporal expression of Tekt1, a mouse tektin C homologue, during spermatogenesis suggest that it is involved in the development of the sperm tail basal body and axoneme. Eur. J. Cell Biol. 79 (2000) 718-725
McKean P.G., Baines A., Vaughan S., and Gull K. Gamma-tubulin functions in the nucleation of a discrete subset of microtubules in the eukaryotic flagellum. Curr. Biol. 13 (2003) 598-602
Weinmann A.S., Yan P.S., Oberley M.J., Huang T.H., and Farnham P.J. Isolating human transcription factor targets by coupling chromatin immunoprecipitation and CpG island microarray analysis. Genes Dev. 16 (2002) 235-244
Whitfield M.L., Sherlock G., Saldanha A.J., Murray J.I., Ball C.A., Alexander K.E., Matese J.C., Perou C.M., Hurt M.M., Brown P.O., and Botstein D. Identification of genes periodically expressed in the human cell cycle and their expression in tumors. Mol. Biol. Cell 13 (2002) 1977-2000
King S.M. Axonemal protofilament ribbons, DM10 domains, and the link to juvenile myoclonic epilepsy. Cell Motil. Cytoskeleton 63 (2006) 245-253
