Abstract
Condensin complexes are thought to play essential roles in mitotic chromosome assembly and segregation in eukaryotes. To date, two condensin complexes (condensin I and II) have been identified. Both complexes contain two structural maintenance of chromosome (SMC) subunits and three non-SMC subunits. In plants, little is known about the localization and function of all the condensin subunits. Here, we report the analyses on the localization of a non-SMC subunit of Arabidopsis condensin I and II, AtCAP-H, and AtCAP-H2, respectively. Our study indicated that localization of AtCAP-H and AtCAP-H2 is dynamically changed through the mitotic cell cycle using GFP-tagged AtCAP-H and AtCAP-H2 in tobacco cultured cells. They are localized at mitotic chromosomes from prometaphase to telophase. However, their localization in interphase is quite different. AtCAP-H was mainly found in the cytoplasm whereas AtCAP-H2 was mainly found in a nucleolus. It is revealed using GFP-tagged deletion mutant s of AtCAP-H that the kleisin-γ middle domain (GM domain) is a unique domain only in AtCAP-H, responsible for chromosomal localization. We propose that the GM domain of CAP-H is essential for its chromosomal localization at mitosis and thus proper function of CAP-H. Differences in localization of AtCAP-H and AtCAP-H2 at interphase also suggest their functional differentiation.
Abbreviations
- CAP:
-
Chromosome Associate Protein
- GFP:
-
Green fluorescent protein
- GM:
-
Kleisin-γ middle
- PEG:
-
Polyethylene glycol
- SMC:
-
Structural maintenance of chromosome
References
Anderson DE, Losada A, Erickson HP, Hirano T (2002) Condensin and cohesin display different arm conformations with characteristic hinge angles. J Cell Biol 156:419–424
Aono N, Sutani T, Tomonaga T, Mochida S, Yanagida M (2002) Cnd2 has dual roles in mitotic condensation and interphase. Nature 417:197–202
Arumugam P, Gruber S, Tanaka K, Haering CH, Mechtler K, Nasmyth K (2003) ATP hydrolysis is required for cohesin’s association with chromosomes. Curr Biol 13:1941–1953
Ball Jr AR, Schmiesing JA, Zhou C, Gregson HC, Okada Y, Doi T and Yokomori K (2002) Identification of a chromosome-targeting domain in the human condensin subunit CNAP1/hCAP-D2/Eg7. Mol Cell Biol 22:5769–5781
Bhat MA, Philp AV, Glover DM, Bellen HJ (1996) Chromatid segregation at anaphase requires the barren product, a novel chromosome-associated protein that interacts with Topoisomerase II. Cell 87:1103–1114
Cabello OA, Eliseeva E, He WG, Youssoufian H, Plon SE, Brinkley BR, Belmont JW (2001) Cell cycle-dependent expression and nucleolar localization of hCAP-H. Mol Biol Cell 12:3527–3537
Freeman L, Aragon-Alcaide L, Strunnikov A (2000) The condensin complex governs chromosome condensation and mitotic transmission of rDNA. J Cell Biol 149:811–824
Fujimoto S, Matsunaga S, Yonemura M, Uchiyama S, Azuma T, Fukui K (2004) Identification of a novel plant MAR DNA binding protein localized an chromosome surfaces. Plant Mol Biol 56:225–239
Gruber S, Haering CH, Nasmyth K (2003) Chromosomal cohesin forms a ring. Cell 112:765–777
Haering CH, Nasmyth K (2003) Building and breaking bridges between sister chromatids. Bioessays 25:1178–1191
Haering CH, Schoffnegger D, Nishino T, Helmhart W, Nasmyth K, Lowe J (2004) Structure and stability of cohesin’s Smc1-Kleisin interaction. Mol Cell 15:951–964
Hagstrom KA, Holmes VF, Cozzarelli NR, Meyer BJ (2002) C. elegans condensin promotes mitotic chromosome architecture, centromere organization, and sister chromatid segregation during mitosis and meiosis. Genes Dev 16:729–742
Hirano T, Mitchison TJ (1994) A heterodimeric coiled-coil protein required for mitotic chromosome condensation in vitro. Cell 79:449–458
Hirano T, Kobayashi R, Hirano M (1997) Condensins, chromosome condensation protein complexes containing XCAP-C, XCAP-E and a Xenopus homolog of the Drosophila Barren protein. Cell 89:511–521
Hirota T, Gerlich D, Koch B, Ellenberg J, Peters JM (2004) Distinct functions of condensin I and II in mitotic chromosome assembly. J Cell Sci 117:6435–6445
Kimura K, Hirano T (2000) Dual roles of the 11S regulatory subcomplex in condensin functions. Proc Natl Acad Sci USA 97:11972–11977
Kimura K, Cuvier O, Hirano T (2001) Chromosome condensation by a human condensin complex in Xenopus egg extracts. J Biol Chem 276:5417–5420
Leon S, Touraine B, Briat JF, Lobreaux S (2002) The AtNFS2 gene from Arabidopsis thaliana encodes a NifS-like plastidial cysteine desulphurase. Biochem J 366:557–564
Liu C, McElver J, Tzafrir I, Joosen R, Wittich P, Patton D, Van Lammeren AA, Meinke D (2002) Condensin and cohesin knockouts in Arabidopsis exhibit a titan seed phenotype. Plant J 29:405–415
Machin F, Paschos K, Jarmuz A, Torres-Rosell J, Pade C, Aragon L (2004) Condensin regulates rDNA silencing by modulating nucleolar Sir2p. Curr Biol 14:125–130
Nagata T, Nemoto Y, Hasegawa S (1992) Tobacco BY-2 cell line as the ‘HeLa’ cell in the cell biology of higher plants. Int Rev Cytol 132:1–30
Neuwald AF, Hirano T (2000) HEAT repeats associated with condensins, cohesins, and other complexes involved in chromosome-related functions. Genome Res 10:1445–1452
Ono T, Losada A, Hirano M, Myers MP, Neuwald AF, Hirano T (2003) Differential contributions of condensin I and condensin II to mitotic chromosome architecture in vertebrate cells. Cell 115:109–121
Ono T, Fang Y, Spector DL, Hirano T (2004) Spatial and temporal regulation of Condensins I and II in mitotic chromosome assembly in human cells. Mol Biol Cell 15:3296–3308
Ratcliffe OJ, Kumimoto RW, Wong BJ, Riechmann JL (2003) Analysis of the Arabidopsis MADS AFFECTING FLOWERING gene family: MAF2 prevents vernalization by short periods of cold. Plant Cell 15:1159–1169
Saitoh N, Goldberg IG, Wood ER, Earnshaw WC (1994). ScII: an abundant chromosome scaffold protein is a member of a family of putative ATPases with an unusual predicted tertiary structure. J Cell Biol 127:303–318
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Schleiffer A, Kaitna S, Maurer-Stroh S, Glotzer M, Nasmyth K, Eisenhaber F (2003) Kleisins: a superfamily of bacterial and eukaryotic SMC protein partners. Mol Cell 11:571–575
Schmiesing JA, Gregson HC, Zhou S, Yokomori K (2000) A human condensin complex containing hCAP-C-hCAP-E and CNAP1, a homolog of Xenopus XCAP-D2, colocalizes with phosphorylated histone H3 during the early stage of mitotic chromosome condensation. Mol Cell Biol 20:6996–7006
Shimizu K, Shirataki H, Honda T, Minami S, Takai Y (1998) Complex formation of SMAP/KAP3, a KIF3A/B ATPase motor-associated protein, with a human chromosome-associated polypeptide. J Biol Chem 273:6591–6594
Siddiqui NU, Stronghill PE, Dengler RE, Hasenkampf CA, Riggs CD (2003) Mutations in Arabidopsis condensin genes disrupt embryogenesis, meristem organization and segregation of homologous chromosomes during meiosis. Development 130:3283–3295
Steffensen S, Coelho PA, Cobbe N, Vass S, Costa M, Hassan B, Prokopenko SN, Bellen H, Heck MMS, Sunkel CE (2001). A role for Drosophila SMC4 in the resolution of sister chromatids in mitosis. Curr Biol 11:295–307
Sutani T, Yuasa T, Tomonaga T, Dohmae N, Takio K, Yanagida M (1999) Fission yeast condensin complex: essential roles of non-SMC subunits for condensation and Cdc2 phosphorylation of Cut3/SMC4. Genes Dev 13:2271–2283
Swedlow JR, Hirano T (2003) The making of the mitotic chromosome: modern insights into classical questions. Mol Cell 11:557–569
Uhlmann F, Wernic D, Poupart MA, Koonin EV, Nasmyth K (2000) Cleavage of cohesin by the CD clan protease separin triggers anaphase in yeast. Cell 103:375–386
Uzbekov R, Timirbulatova E, Watrin E, Cubizolles F, Ogereau D, Gulak P, Legagneux V, Polyakov VJ, Le Guellec K, Kireev I (2003) Nucleolar association of pEg7 and XCAP-E, two members of Xenopus laevis condensin complex in interphase cells. J Cell Sci 116:1667–1678
Wignall SM, Deehan R, Maresca TJ, Heald R (2003) The condensin complex is required for proper spindle assembly and chromosome segregation in Xenopus egg extracts. J Cell Biol 161:1041–1051
Yeong FM, Hombauer H, Wendt KS, Hirota T, Mudrak I, Mechtler K, Loregger T, Marchler-Bauer A, Tanaka K, Peters JM, Ogris E (2003) Identification of a subunit of a novel Kleisin-beta/SMC complex as a potential substrate of protein phosphatase 2A. Curr Biol 13:2058–2064
Acknowledgements
We thank Akira Kawabe for valuable discussions and Joyce A. Cartagena and Reiko Isobe for technical assistances. This study was supported by a fund from the Ministry of Education, Culture, Sports, Science and Technology, Japan, to Kiichi Fukui and the Industrial Technology Research Grant Program in 2003 from the New Energy and Technology Development Organization (NEDO) of Japan.
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Fujimoto, S., Yonemura, M., Matsunaga, S. et al. Characterization and dynamic analysis of Arabidopsis condensin subunits, AtCAP-H and AtCAP-H2. Planta 222, 293–300 (2005). https://doi.org/10.1007/s00425-005-1546-0
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DOI: https://doi.org/10.1007/s00425-005-1546-0