DOI: 10.1091/mbc.e03-12-0888. Role of the Midbody Matrix in Cytokinesis: RNAi and Genetic Rescue Analysis of the Mammalian Motor Protein CHO1

Role of the Midbody Matrix in Cytokinesis: RNAi and Genetic Rescue Analysis of the Mammalian Motor Protein CHO1

10.1091/mbc.e03-12-0888

Crossref journal-article

American Society for Cell Biology (ASCB)

Molecular Biology of the Cell (1076)

Abstract

CHO1 is a kinesin-like motor protein essential for cytokinesis in mammalian cells. To analyze how CHO1 functions, we established RNAi and genetic rescue assays. CHO1-depleted cells reached a late stage of cytokinesis but fused back to form binucleate cells because of the absence of the midbody matrix in the middle of the intercellular bridge. Expression of exogenous CHO1 restored the formation of the midbody matrix and rescued cytokinesis in siRNA-treated cells. By analyzing phenotypes rescued with different constructs, it was shown that both motor and stalk domains function in midbody formation, whereas the tail is essential for completion of cytokinesis after the midbody matrix has formed. During the terminal stage of cytokinesis, different subregions of the tail play distinctive roles in stabilizing the midbody matrix and maintaining an association between the midbody and cell cortex. These results demonstrate that CHO1 consists of functionally differentiated subregions that act in concert to ensure complete cell separation.

Bibliography

Matuliene, J., & Kuriyama, R. (2004). Role of the Midbody Matrix in Cytokinesis: RNAi and Genetic Rescue Analysis of the Mammalian Motor Protein CHO1. Molecular Biology of the Cell, 15(7), 3083â3094.

Authors 2

Jurgita Matuliene (first)
Ryoko Kuriyama (additional)

References 49 Referenced 44

Dates

Type	When
Created	21 years, 4 months ago (April 12, 2004, 8:53 p.m.)
Deposited	4 years, 2 months ago (June 15, 2021, 10:45 a.m.)
Indexed	2 months ago (July 1, 2025, 1:25 p.m.)
Issued	21 years, 2 months ago (July 1, 2004)
Published	21 years, 2 months ago (July 1, 2004)
Published Print	21 years, 2 months ago (July 1, 2004)

Funders 0

None

BibTeX

@article{Matuliene_2004, title={Role of the Midbody Matrix in Cytokinesis: RNAi and Genetic Rescue Analysis of the Mammalian Motor Protein CHO1}, volume={15}, ISSN={1939-4586}, url={http://dx.doi.org/10.1091/mbc.e03-12-0888}, DOI={10.1091/mbc.e03-12-0888}, number={7}, journal={Molecular Biology of the Cell}, publisher={American Society for Cell Biology (ASCB)}, author={Matuliene, Jurgita and Kuriyama, Ryoko}, year={2004}, month=jul, pages={3083–3094} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        7,
        1
      ]
    ],
    "date-time": "2025-07-01T17:25:25Z",
    "timestamp": 1751390725667
  },
  "reference-count": 49,
  "publisher": "American Society for Cell Biology (ASCB)",
  "issue": "7",
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        2004,
        7
      ]
    ]
  },
  "abstract": "<jats:p>CHO1 is a kinesin-like motor protein essential for cytokinesis in mammalian cells. To analyze how CHO1 functions, we established RNAi and genetic rescue assays. CHO1-depleted cells reached a late stage of cytokinesis but fused back to form binucleate cells because of the absence of the midbody matrix in the middle of the intercellular bridge. Expression of exogenous CHO1 restored the formation of the midbody matrix and rescued cytokinesis in siRNA-treated cells. By analyzing phenotypes rescued with different constructs, it was shown that both motor and stalk domains function in midbody formation, whereas the tail is essential for completion of cytokinesis after the midbody matrix has formed. During the terminal stage of cytokinesis, different subregions of the tail play distinctive roles in stabilizing the midbody matrix and maintaining an association between the midbody and cell cortex. These results demonstrate that CHO1 consists of functionally differentiated subregions that act in concert to ensure complete cell separation.</jats:p>",
  "DOI": "10.1091/mbc.e03-12-0888",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2004,
        4,
        13
      ]
    ],
    "date-time": "2004-04-13T00:53:37Z",
    "timestamp": 1081817617000
  },
  "page": "3083-3094",
  "source": "Crossref",
  "is-referenced-by-count": 44,
  "title": "Role of the Midbody Matrix in Cytokinesis: RNAi and Genetic Rescue Analysis of the Mammalian Motor Protein CHO1",
  "prefix": "10.1091",
  "volume": "15",
  "author": [
    {
      "given": "Jurgita",
      "family": "Matuliene",
      "sequence": "first",
      "affiliation": [
        {
          "name": "Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota 55455"
        }
      ]
    },
    {
      "given": "Ryoko",
      "family": "Kuriyama",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota 55455"
        }
      ]
    }
  ],
  "member": "1076",
  "reference": [
    {
      "key": "REF1",
      "doi-asserted-by": "crossref",
      "unstructured": "Abaza, A., Soleilhac, J.M., Westendorf, J., Piel, M. Crevel, I., Roux, A., and Pirollet, F. (2003). M phase phosphoprotein 1 is a human plus-end-directed kinesin-related protein required for cytokinesis.J. Biol. Chem.278,27844-27852.",
      "DOI": "10.1074/jbc.M304522200"
    },
    {
      "key": "REF2",
      "doi-asserted-by": "crossref",
      "unstructured": "Adams, R.R., Tavares, A.A.M., Salzberg, A., Bellen, H.J., and Glover, D.M. (1998).pavarottiencodes a kinesin-like protein required to organize the central spindle and contractile ring for cytokinesis.Genes Dev.12,1483-1494.",
      "DOI": "10.1101/gad.12.10.1483"
    },
    {
      "key": "REF3",
      "doi-asserted-by": "crossref",
      "unstructured": "Alsop, G.B., and Zhang, D. (2003). Microtubules are the only structural constituent of the spindle apparatus required for induction of cell cleavage.J. Cell Biol.162,383-390.",
      "DOI": "10.1083/jcb.200301073"
    },
    {
      "key": "REF4",
      "unstructured": "Boman, A.L., and Kahn, R.A. (1995). Arf proteins: the membrane traffic police?Trends Biochem. Sci.20,147-150."
    },
    {
      "key": "REF5",
      "doi-asserted-by": "crossref",
      "unstructured": "Boman, A.L., J. Kuai, J., Zhu, X., Chen, J., Kuriyama, R., and Kahn, R.A. (1999). Arf proteins bind to mitotic kinesin-like protein 1 (MKLP1) in a GTP-dependent fashion.Cell Motil. Cytoskel.44,119-132.",
      "DOI": "10.1002/(SICI)1097-0169(199910)44:2<119::AID-CM4>3.0.CO;2-C"
    },
    {
      "key": "REF6",
      "doi-asserted-by": "crossref",
      "unstructured": "Buck, R.C., and Tisdale, J.M. (1962). The fine structure of the mid-body of the rat erythroblast.J. Cell Biol.13,109-115.",
      "DOI": "10.1083/jcb.13.1.109"
    },
    {
      "key": "REF7",
      "doi-asserted-by": "publisher",
      "DOI": "10.1091/mbc.7.2.225"
    },
    {
      "key": "REF8",
      "doi-asserted-by": "crossref",
      "unstructured": "Chen, M.C., Zhou, Y., and Detrich, H.W., III. (2002). Zebrafish mitotic kinesin-like protein 1 (Mklp1) functions in embryonic cytokinesis.Physiol. Genomics8,51-66.",
      "DOI": "10.1152/physiolgenomics.00042.2001"
    },
    {
      "key": "REF9",
      "doi-asserted-by": "crossref",
      "unstructured": "Dechant, R., and Glotzer, M. (2003). Centrosome separation and central spindle assembly act in redundant pathways that regulate microtubule density and trigger cleavage furrow formation.Dev. Cell4,333-344.",
      "DOI": "10.1016/S1534-5807(03)00057-1"
    },
    {
      "key": "REF10",
      "doi-asserted-by": "crossref",
      "unstructured": "Donaldson, J.G., Radhakrishna, H., and Peters, P.J. (1995). The ARF GTPases: defining roles in membrane traffic and organelle structure. Cold Spring Harb.Symp. Quant. Biol.60,229-234.",
      "DOI": "10.1101/SQB.1995.060.01.026"
    },
    {
      "key": "REF11",
      "doi-asserted-by": "crossref",
      "unstructured": "Donaldson, J.G. (2003). Multiple roles for Arf6: sorting, structuring, and signaling at the plasma membrane.J. Biol. Chem.278,41573-41576.",
      "DOI": "10.1074/jbc.R300026200"
    },
    {
      "key": "REF12",
      "doi-asserted-by": "crossref",
      "unstructured": "Giansanti, M.G., Bonaccorsi, S., Williams, B., Williams, E.V., Santolamazza, C., Goldberg, M.K., and Gatti, M. (1998). Cooperative interactions between the central spindle and the contractile ring duringDrosophilacytokinesis.Genes Dev.12,396-410.",
      "DOI": "10.1101/gad.12.3.396"
    },
    {
      "key": "REF13",
      "doi-asserted-by": "crossref",
      "unstructured": "Hill, E., Clarke, M., and Barr, F.A. (2000). The Rab6-binding kinesin, Rab6-KIFL, is required for cytokinesis.EMBO J.19,5711-5719.",
      "DOI": "10.1093/emboj/19.21.5711"
    },
    {
      "key": "REF14",
      "doi-asserted-by": "crossref",
      "unstructured": "Hirose, K., Kawashima, T., Iwamoto, I., Nosaka, T., and Kitamura, T. (2001). MgcRacGAP is involved in cytokinesis through associating with mitotic spindle and midbody.J. Biol. Chem.276,5821-5828.",
      "DOI": "10.1074/jbc.M007252200"
    },
    {
      "key": "REF15",
      "doi-asserted-by": "crossref",
      "unstructured": "Jantsch-Plunger, V., Gonczy, P., Romano, A., Schnabel, H., Hamill, D., Schnabel, R., Hyman, A.A., and Glotzer, M. (2000). CYK-4, a Rho family GTPase activating protein (GAP) required for central spindle formation and cytokinesis.J. Cell Biol.149,1391-1404.",
      "DOI": "10.1083/jcb.149.7.1391"
    },
    {
      "key": "REF16",
      "doi-asserted-by": "crossref",
      "unstructured": "Jiang, W., Jimenez, G., Wells, N.J., Hope, T.J., Wahl, G.M., Hunter, T., and Fukunaga, R. (1998). PRC1, a human mitotic spindle-associated CDK substrate protein required for cytokinesis.Mol. Cell2,877-885.",
      "DOI": "10.1016/S1097-2765(00)80302-0"
    },
    {
      "key": "REF17",
      "doi-asserted-by": "crossref",
      "unstructured": "King, R.C., and Akai, H. (1971). Spermatogenesis inBombyx mori.I. The canal system joining sister spermatocytes.J. Morphol.134,47-56.",
      "DOI": "10.1002/jmor.1051340104"
    },
    {
      "key": "REF18",
      "doi-asserted-by": "crossref",
      "unstructured": "Kuriyama, R., Dragas-Granoic, S., Maekawa, T., Vassilev, A., Khodjakov, A., and Kobayashi, H. (1994). Heterogeneity and microtubule interaction of the CHO1 antigen, a mitosis-specific kinesin-like protein. Analysis of subdomains expressed in insect Sf9 cells.J. Cell Sci.107,3485-3499.",
      "DOI": "10.1242/jcs.107.12.3485"
    },
    {
      "key": "REF19",
      "doi-asserted-by": "crossref",
      "unstructured": "Kuriyama, R., Gustus, C., Uetake, Y., Terada, Y., and Matuliene, J. (2002). CHO1, a mammalian kinesin-like protein, interacts with F-actin and is involved in the terminal phase of cytokinesis.J. Cell Biol.156,783-790.",
      "DOI": "10.1083/jcb.200109090"
    },
    {
      "key": "REF20",
      "doi-asserted-by": "crossref",
      "unstructured": "Matuliene, J., R. Essner, R., Ryu, J.H., Hamaguchi, Y., Baas, P.W., Haraguchi, T., Hiraoka, Y., and Kuriyama, R. (1999). Function of a minus-end-directed kinesin-like motor protein in mammalian cells.J. Cell Sci.112,4041-4050.",
      "DOI": "10.1242/jcs.112.22.4041"
    },
    {
      "key": "REF21",
      "doi-asserted-by": "publisher",
      "DOI": "10.1091/mbc.01-10-0504"
    },
    {
      "key": "REF22",
      "doi-asserted-by": "crossref",
      "unstructured": "McIntosh, J.R., and Landis, S.C. (1971). The distribution of spindle microtubules during mitosis in cultured mammalian cells.J. Cell Biol.49,468-497.",
      "DOI": "10.1083/jcb.49.2.468"
    },
    {
      "key": "REF23",
      "doi-asserted-by": "crossref",
      "unstructured": "Miki, H., Setou, M., Kaneshiro, K., and Hirokawa, N. (2001). All kinesin superfamily protein, KLF, genes in mouse and human.Proc. Natl. Acad. Sci. USA98,7004-7011.",
      "DOI": "10.1073/pnas.111145398"
    },
    {
      "key": "REF24",
      "doi-asserted-by": "publisher",
      "DOI": "10.1091/mbc.e03-04-0214"
    },
    {
      "key": "REF25",
      "doi-asserted-by": "crossref",
      "unstructured": "Minestrini, G., Mathe, E., and Glover, D.M. (2002). Domains of the Pavarotti kinesin-like protein that direct its subcellular distribution: effects of mislocalisation on the tubulin and actin cytoskeleton duringDrosophilaoogenesis.J. Cell Sci.115,725-736.",
      "DOI": "10.1242/jcs.115.4.725"
    },
    {
      "key": "REF26",
      "doi-asserted-by": "crossref",
      "unstructured": "Minoshima Y.et al.(2003). Phosphorylation by aurora B converts MgcRacGAP to a RhoGAP during cytokinesis.Dev. Cell4,549-560.",
      "DOI": "10.1016/S1534-5807(03)00089-3"
    },
    {
      "key": "REF27",
      "doi-asserted-by": "crossref",
      "unstructured": "Mishima, M., Kaitna, S., and Glotzer, M. (2002). Central spindle assembly and cytokinesis require a kinesin-like protein/RhoGAP complex with microtubule bundling activity.Dev. Cell2,41-54.",
      "DOI": "10.1016/S1534-5807(01)00110-1"
    },
    {
      "key": "REF28",
      "doi-asserted-by": "crossref",
      "unstructured": "Mollinari, C., Kleman, J.P., Jiang, W., Schoehn, G., Hunter, T., and Margolis, R.L. (2002). PRC1 is a microtubule binding and bundling protein essential to maintain the mitotic spindle midzone.J. Cell Biol.157,1175-1186.",
      "DOI": "10.1083/jcb.200111052"
    },
    {
      "key": "REF29",
      "doi-asserted-by": "crossref",
      "unstructured": "Mullins, J.M., and Biesele, J.J. (1977). Terminal phase of cytokinesis in D-98s cells.J. Cell Biol.73,672-684.",
      "DOI": "10.1083/jcb.73.3.672"
    },
    {
      "key": "REF30",
      "doi-asserted-by": "crossref",
      "unstructured": "Mullins, J.M., and McIntosh, J.R. (1982). Isolation and initial characterization of the mammalian midbody.J. Cell Biol.94,654-661.",
      "DOI": "10.1083/jcb.94.3.654"
    },
    {
      "key": "REF31",
      "doi-asserted-by": "crossref",
      "unstructured": "Neef, R., Preisinger, C., Sutcliffe, J., Kopajtich, R., Nigg, E.A., Mayer, T.U., and Barr, F.A. (2003). Phosphorylation of mitotic kinesin-like protein 2 by polo-like kinase 1 is required for cytokinesis.J. Cell Biol.162,863-876.",
      "DOI": "10.1083/jcb.200306009"
    },
    {
      "key": "REF32",
      "doi-asserted-by": "crossref",
      "unstructured": "Nislow, C., Lombillo, V.A., Kuriyama, R., and McIntosh, J.R. (1992). A plus-end-directed motor enzyme that moves antiparallel microtubules in vitro localizes to the interzone of mitotic spindles.Nature359,543-547.",
      "DOI": "10.1038/359543a0"
    },
    {
      "key": "REF33",
      "doi-asserted-by": "crossref",
      "unstructured": "Norman, J.C., Jones, D., Barry, S.T., Holt, M.R., Cockcroft, S., and Critchley, D.R. (1998). ARF1 mediates paxillin recruitment to focal adhesions and potentiates Rho-stimulated stress fiber formation in intact and permeabilized Swiss 3T3 fibroblasts.J. Cell Biol.143,1981-1995.",
      "DOI": "10.1083/jcb.143.7.1981"
    },
    {
      "key": "REF34",
      "doi-asserted-by": "crossref",
      "unstructured": "Powers, J., Bossinger, O., Rose, D., Strome, S., and Saxton, W. (1998). A nematode kinesin required for cleavage furrow advancement.Curr. Biol.8,1133-1136.",
      "DOI": "10.1016/S0960-9822(98)70470-1"
    },
    {
      "key": "REF35",
      "doi-asserted-by": "crossref",
      "unstructured": "Quimby, B.B., and Dasso, M. (2003). The small GTPase Ran: interpreting the signs.Curr. Opin. Cell Biol.15,338-344.",
      "DOI": "10.1016/S0955-0674(03)00046-2"
    },
    {
      "key": "REF36",
      "doi-asserted-by": "publisher",
      "DOI": "10.1091/mbc.9.8.2037"
    },
    {
      "key": "REF37",
      "doi-asserted-by": "crossref",
      "unstructured": "Rappaport, R. (1961). Experiments concerning the cleavage stimulus in sand dollar eggs.J. Exp. Zool.148,81-89.",
      "DOI": "10.1002/jez.1401480107"
    },
    {
      "key": "REF38",
      "doi-asserted-by": "crossref",
      "unstructured": "Rappaport, R., and Rappaport, B.N. (1974). Establishment of cleavage furrows by the mitotic spindle.J. Exp. Zool.189,189-196.",
      "DOI": "10.1002/jez.1401890206"
    },
    {
      "key": "REF39",
      "doi-asserted-by": "crossref",
      "unstructured": "Rappaport, R., and Rappaport, B.N. (1983). Cytokinesis: effects of blocks between the mitotic apparatus and the surface on furrow establishment in flattened echinoderm eggs.J. Exp. Zool.227,213-227.",
      "DOI": "10.1002/jez.1402270206"
    },
    {
      "key": "REF40",
      "doi-asserted-by": "crossref",
      "unstructured": "Rappaport, R. (1986). Establishment of the mechanism of cytokinesis in animal cells.Int. Rev. Cytol.105,245-281.",
      "DOI": "10.1016/S0074-7696(08)61065-7"
    },
    {
      "key": "REF41",
      "doi-asserted-by": "crossref",
      "unstructured": "Salmon, E.D., Goode, D., Maugel, T.K., and Bonar, B.D. (1976). Pressure-induced depolymerization of spindle microtubules. III. Differential stability in HeLa cells.J. Cell Biol.69,443-454.",
      "DOI": "10.1083/jcb.69.2.443"
    },
    {
      "key": "REF42",
      "doi-asserted-by": "crossref",
      "unstructured": "Sellitto, C., and Kuriyama, R. (1988). Distribution of a matrix component of the midbody during the cell cycle in Chinese hamster ovary cells.J. Cell Biol.106,431-439.",
      "DOI": "10.1083/jcb.106.2.431"
    },
    {
      "key": "REF43",
      "doi-asserted-by": "crossref",
      "unstructured": "Severson, A.F., Hamill, D.R., Carter, J.C., Schumacher, J., and Bowerman, B. (2000). The Aurora-related kinase AIR-2 recruits ZEN-4/CeMKLP1 to the mitotic spindle at metaphase and is required for cytokinesis.Curr. Biol.10,1162-1171.",
      "DOI": "10.1016/S0960-9822(00)00715-6"
    },
    {
      "key": "REF44",
      "doi-asserted-by": "crossref",
      "unstructured": "Somers, W.G., and Saint, R. (2003). A RhoGEF and Rho family GTPase-activating protein complex links the contractile ring to cortical microtubules at the onset of cytokinesis.Dev. Cell4,29-39.",
      "DOI": "10.1016/S1534-5807(02)00402-1"
    },
    {
      "key": "REF45",
      "doi-asserted-by": "publisher",
      "DOI": "10.1091/mbc.01-12-0589"
    },
    {
      "key": "REF46",
      "doi-asserted-by": "crossref",
      "unstructured": "Skop, A.R., Bergmann, D., Mohler, W.A., and White, J.G. (2001). Completion of cytokinesis inC. elegansrequires a brefeldin A-sensitive membrane accumulation at the cleavage furrow apex.Curr. Biol.11,735-746.",
      "DOI": "10.1016/S0960-9822(01)00231-7"
    },
    {
      "key": "REF47",
      "unstructured": "Skop, A.R., Liu, H., Yates, J., Meyer, B.J., and Heald, R. (2002). Functional proteomic analysis of the mammalian midbody reveals conserved cell division components.Mol. Biol. Cell13,3a."
    },
    {
      "key": "REF48",
      "doi-asserted-by": "crossref",
      "unstructured": "Tatsumoto, T., Xie, X., Blumenthal, R., Okamoto, I., and Miki, T. (1999). Human ECT2 is an exchange factor for Rho GTPases, phosphorylated in G2/M phase, and involved in cytokinesis.J. Cell Biol.147,921-927.",
      "DOI": "10.1083/jcb.147.5.921"
    },
    {
      "key": "REF49",
      "doi-asserted-by": "crossref",
      "unstructured": "Wheatley, S.P., and Wang, Y. (1996). Midzone microtubule bundles are continuously required for cytokinesis in cultured epithelial cells.J. Cell Biol.135,981-989.",
      "DOI": "10.1083/jcb.135.4.981"
    }
  ],
  "container-title": "Molecular Biology of the Cell",
  "original-title": [],
  "language": "en",
  "deposited": {
    "date-parts": [
      [
        2021,
        6,
        15
      ]
    ],
    "date-time": "2021-06-15T14:45:49Z",
    "timestamp": 1623768349000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://www.molbiolcell.org/doi/10.1091/mbc.e03-12-0888"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2004,
        7
      ]
    ]
  },
  "references-count": 49,
  "journal-issue": {
    "issue": "7",
    "published-print": {
      "date-parts": [
        [
          2004,
          7
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1091/mbc.e03-12-0888"
  ],
  "URL": "http://dx.doi.org/10.1091/mbc.e03-12-0888",
  "relation": {
    "has-review": [
      {
        "id-type": "doi",
        "id": "10.3410/f.1019788.222594",
        "asserted-by": "object"
      }
    ]
  },
  "ISSN": [
    "1059-1524",
    "1939-4586"
  ],
  "subject": [],
  "container-title-short": "MBoC",
  "published": {
    "date-parts": [
      [
        2004,
        7
      ]
    ]
  }
}