Abstract
The INCENP (inner centromere protein) is a chromosomal passenger protein that plays multiple roles in regulating mitosis and cytokinesis. The MKLP1 (mitotic kinesin-like protein) is a component of centralspindlin complex that has been implicated in assembly of midzone/midbody during mitosis and is essential for cytokinesis. In the present study, we investigated functions of INCNEP and MKLP1 and their interplay in regulating spindle midzone/midbody formation and cytokinesis in human cells. Immunofluorescence and live-cell imaging analyses have shown that, in addition to multiple chromosome segregation defects, cells that lacked INCENP by RNAi (RNA interference) exhibit abnormal spindle midzone/midbody formation, resulting in formation of binucleated/multinucleated cells. Suppression of MKLP1 expression by siRNA (small interfering RNA) did not cause any abnormality of chromosome segregation and midzone formation, but abrogated midbody formation and completion of cytokinesis. Furthermore, we show that INCENP is required for recruiting MKLP1 to the spindle midzone/midbody. Three-dimensional reconstruction imaging analysis suggests that recruitment of MKLP1 to the midzone/midbody by INCENP is a crucial step for the midbody formation and completion of cytokinesis in mammalian cells.
References
52
Referenced
78
10.1083/jcb.200310112
/ J. Cell Biol. / Cleavage furrow positioning by Glotzer (2004)10.1083/jcb.138.2.385
/ J. Cell Biol. / CDK1 inactivation regulates anaphase spindle dynamics and cytokinesis in vivo by Wheatley (1997)10.1016/S0962-8924(00)01880-8
/ Trends Cell Biol. / Chromosomal passengers and the (aurora) ABCs of mitosis by Adams (2001)10.1016/S1534-5807(01)00110-1
/ Dev. Cell. / Central spindle assembly and cytokinesis require a kinesin-like protein/RhoGAP complex with microtubule bundling activity by Mishima (2002)10.1083/jcb.200111052
/ J. Cell Biol. / PRC1 is a microtubule binding and bundling protein essential to maintain the mitotic spindle midzone by Mollinari (2002)10.1091/mbc.e03-12-0888
/ Mol. Biol. Cell / Role of the midbody matrix in cytokinesis: RNAi and genetic rescue analysis of the mammalian motor protein CHO1 by Matuliene (2004)10.1083/jcb.200403084
/ J. Cell Biol. / Relocation of Aurora B from centromeres to the central spindle at the metaphase to anaphase transition requires MKlp2 by Gruneberg (2004)10.1038/nature02767
/ Nature (London) / Cell cycle regulation of central spindle assembly by Mishima (2004)10.1083/jcb.200303022
/ J. Cell Biol. / The roles of microtubule-based motor proteins in mitosis: comprehensive RNAi analysis in the Drosophila S2 cell line by Goshima (2003)10.1073/pnas.0408438102
/ Proc. Natl. Acad. Sci. U.S.A. / Cell-cycle-dependent translocation of PRC1 on the spindle by Kif4 is essential for midzone formation and cytokinesis by Zhu (2005)10.1101/SQB.1993.058.01.077
/ Quant. Biol. / The INCENPs: structural and functional analysis of a family of chromosome passenger proteins. Cold Spring Harbor Symp by Mackay (1993)10.1083/jcb.151.7.1575
/ J. Cell Biol. / Human survivin is a kinetochore-associated passenger protein by Skoufias (2000)10.1247/csf.26.653
/ Cell Struct. Funct. / Role of chromosomal passenger complex in chromosome segregation and cytokinesis by Terada (2001)10.1038/426780a
/ Nature (London) / Cell cycle: passenger acrobatics by Higuchi (2003)10.1016/S1534-5807(03)00205-3
/ Dev. Cell. / The mammalian passenger protein TD-60 is an RCC1 family member with an essential role in prometaphase to metaphase progression by Mollinari (2003)10.1083/jcb.200207117
/ J. Cell Biol. / CSC-1: a subunit of the Aurora B kinase complex that binds to the survivin-like protein BIR-1 and the incenp-like protein ICP-1 by Romano (2003)10.1083/jcb.200404001
/ J. Cell Biol. / Borealin: a novel chromosomal passenger required for stability of the bipolar mitotic spindle by Gassmann (2004)10.1016/j.cell.2004.06.026
/ Cell / The chromosomal passenger complex is required for chromatin-induced microtubule stabilization and spindle assembly by Sampath (2004)10.1006/excr.2000.5088
/ Exp. Cell Res. / INCENP binds directly to tubulin and requires dynamic microtubules to target to the cleavage furrow by Wheatley (2001)10.1091/mbc.e02-11-0769
/ Mol. Biol. Cell / Exploring the functional interactions between Aurora B, INCENP, and survivin in mitosis by Honda (2003)10.1007/s00412-004-0307-3
/ Chromosoma / Chromosomal passengers: the four-dimensional regulation of mitotic events by Vagnarelli (2004)10.1083/jcb.140.5.991
/ J. Cell Biol. / A dominant mutant of inner centromere protein (INCENP), a chromosomal protein, disrupts prometaphase congression and cytokinesis by Mackay (1998)10.1083/jcb.143.6.1635
/ J. Cell Biol. / AIR-2: an Aurora/Ipl1-related protein kinase associated with chromosomes and midbody microtubules is required for polar body extrusion and cytokinesis in Caenorhabditis elegans embryos by Schumacher (1998)10.1016/S0960-9822(00)00715-6
/ Curr. Biol. / The aurora-related kinase AIR-2 recruits ZEN-4/CeMKLP1 to the mitotic spindle at metaphase and is required for cytokinesis by Severson (2000)10.1083/jcb.152.4.669
/ J. Cell Biol. / Drosophila aurora B kinase is required for histone H3 phosphorylation and condensin recruitment during chromosome condensation and to organize the central spindle during cytokinesis by Giet (2001)10.1016/S0960-9822(02)00820-5
/ Curr. Biol. / The aurora B kinase AIR-2 regulates kinetochores during mitosis and is required for separation of homologous chromosomes during meiosis by Kaitna (2002)10.1083/jcb.200208091
/ J. Cell Biol. / Aurora B couples chromosome alignment with anaphase by targeting BubR1, Mad2, and Cenp-E to kinetochores by Ditchfield (2003)10.1083/jcb.200208092
/ J. Cell Biol. / The small molecule Hesperadin reveals a role for Aurora B in correcting kinetochore-microtubule attachment and in maintaining the spindle assembly checkpoint by Hauf (2003)10.1007/s003359901014
/ Mamm. Genome / Cloning, expression, and promoter structure of a mammalian inner centromere protein (INCENP) by Saffery (1999)10.1016/S0960-9822(00)00769-7
/ Curr. Biol. / Survivin and the inner centromere protein INCENP show similar cell-cycle localization and gene knockout phenotype by Uren (2000)10.1016/S1534-5807(03)00089-3
/ Dev. Cell / Phosphorylation by aurora B converts MgcRacGAP to a RhoGAP during cytokinesis by Minoshima (2003)10.1016/S1097-2765(03)00151-5
/ Mol. Cell / Closing the GAP: a role for a RhoA GAP in cytokinesis by Maddox (2003)10.1083/jcb.153.4.865
/ J. Cell Biol. / Essential roles of Drosophila inner centromere protein (INCENP) and aurora B in histone H3 phosphorylation, metaphase chromosome alignment, kinetochore disjunction, and chromosome segregation by Adams (2001)10.1002/(SICI)1097-0029(20000415)49:2<202::AID-JEMT13>3.0.CO;2-8
/ Microsc. Res. Tech. / Relationships between the central spindle and the contractile ring during cytokinesis in animal cells by Gatti (2000)10.1146/annurev.cellbio.17.1.351
/ Annu. Rev. Cell Dev. Biol. / Animal cell cytokinesis by Glotzer (2001)10.1128/MMBR.66.2.155-178.2002
/ Microbiol. Mol. Biol. Rev. / Cytokinesis in eukaryotes by Guertin (2002)10.1083/jcb.111.2.511
/ J. Cell Biol. / A monoclonal antibody to a mitotic microtubule-associated protein blocks mitotic progression by Nislow (1990)10.1083/jcb.200109090
/ J. Cell Biol. / CHO1, a mammalian kinesin-like protein, interacts with F-actin and is involved in the terminal phase of cytokinesis by Kuriyama (2002)10.1091/mbc.01-10-0504
/ Mol. Biol. Cell. / Kinesin-like protein CHO1 is required for the formation of midbody matrix and the completion of cytokinesis in mammalian cells by Matuliene (2002)10.1016/S1097-2765(00)80302-0
/ Mol. Cell / PRC1: a human mitotic spindle-associated CDK substrate protein required for cytokinesis by Jiang (1998)10.1091/mbc.12.4.971
/ Mol. Biol. Cell / Cell cycle-dependent changes in microtubule dynamics in living cells expressing green fluorescent protein-α tubulin by Rusan (2001)10.1016/S0960-9822(98)70156-3
/ Curr. Biol. / Histone–GFP fusion protein enables sensitive analysis of chromosome dynamics in living mammalian cells by Kanda (1998)10.1073/pnas.152327299
/ Proc. Natl. Acad. Sci. U.S.A. / Short RNA duplexes produced by hydrolysis with Escherichia coli RNase III mediate effective RNA interference in mammalian cells by Yang (2002)10.1101/gad.862301
/ Genes Dev. / RNA interference is mediated by 21- and 22-nucleotide RNAs by Elbashir (2001)10.1007/s004120100130
/ Chromosoma / Human INCENP colocalizes with the Aurora-B/AIRK2 kinase on chromosomes and is overexpressed in tumour cells by Adams (2001)10.1016/S0960-9822(01)00238-X
/ Curr. Biol. / INCENP is required for proper targeting of Survivin to the centromeres and the anaphase spindle during mitosis by Wheatley (2001)10.1016/S0960-9822(00)00721-1
/ Curr. Biol. / Incenp and an aurora-like kinase form a complex essential for chromosome segregation and efficient completion of cytokinesis by Kaitna (2000)10.1101/gad.12.10.1483
/ Genes Dev. / Pavarotti encodes a kinesin-like protein required to organize the central spindle and contractile ring for cytokinesis by Adams (1998)10.1083/jcb.149.7.1391
/ J. Cell Biol. / CYK-4: A Rho family GTPase activating protein (GAP) required for central spindle formation and cytokinesis by Jantsch-Plunger (2000)10.1016/j.cub.2004.10.040
/ Curr. Biol. / Cytokinesis: the central spindle takes center stage by McCollum (2004)10.1152/physiolgenomics.00042.2001
/ Physiol. Genomics / Zebrafish mitotic kinesin-like protein 1 (Mklp1) functions in embryonic cytokinesis by Chen (2002)10.1091/mbc.e03-04-0214
/ Mol. Biol. Cell / Localization of Pavarotti-KLP in living Drosophila embryos suggests roles in reorganizing the cortical cytoskeleton during the mitotic cycle by Minestrini (2003)
Dates
Type | When |
---|---|
Created | 20 years, 1 month ago (July 12, 2005, 2:35 p.m.) |
Deposited | 3 years, 9 months ago (Nov. 22, 2021, 9:05 p.m.) |
Indexed | 4 months, 4 weeks ago (April 4, 2025, 2:44 a.m.) |
Issued | 20 years, 1 month ago (July 5, 2005) |
Published | 20 years, 1 month ago (July 5, 2005) |
Published Online | 20 years, 1 month ago (July 5, 2005) |
Published Print | 20 years, 1 month ago (July 15, 2005) |
@article{Zhu_2005, title={Recruitment of MKLP1 to the spindle midzone/midbody by INCENP is essential for midbody formation and completion of cytokinesis in human cells}, volume={389}, ISSN={1470-8728}, url={http://dx.doi.org/10.1042/bj20050097}, DOI={10.1042/bj20050097}, number={2}, journal={Biochemical Journal}, publisher={Portland Press Ltd.}, author={Zhu, Changjun and Bossy-Wetzel, Ella and Jiang, Wei}, year={2005}, month=jul, pages={373–381} }