CYK4 promotes antiparallel microtubule bundling by optimizing MKLP1 neck conformation. Davies, T., Kodera, N., Kaminski Schierle, G., S., Rees, E., Erdelyi, M., Kaminski, C., F., Ando, T., Mishima, M., & Gould, K., L. PLoS Biology, 13(4):e1002121, Public Library of Science, 2015. ![CYK4 promotes antiparallel microtubule bundling by optimizing MKLP1 neck conformation [pdf]](https://bibbase.org/img/filetypes/pdf.svg "pdf")
Paper abstract bibtex Centralspindlin, a constitutive 2:2 heterotetramer of MKLP1 (a kinesin-6) and the non-motor subunit CYK4, plays important roles in cytokinesis. It is crucial for the formation of central spindle microtubule bundle structure. Its accumulation at the central antiparallel overlap zone is key for recruitment and regulation of downstream cytokinesis factors and for stable anchoring of the plasma membrane at the midbody. Both MKLP1 and CYK4 are required for efficient microtubule bundling. However, the mechanism by which CYK4 contributes to this is unclear. Here we performed structural and functional analyses of centralspindlin using high-speed atomic force microscopy, Förster resonance energy transfer analysis, and in vitro reconstitution. Our data reveal that CYK4 binds to a globular mass in the atypi-cally long MKLP1 neck domain between the catalytic core and the coiled coil and thereby reconfigures the two motor domains in the MKLP1 dimer to be suitable for antiparallel micro-tubule bundling. Our work provides insights into the microtubule bundling during cytokinesis and into the working mechanisms of the kinesins with non-canonical neck structures. Author Summary Cell division requires coordination of many different cellular components. Cytokinesis is the process by which the cytoplasm divides between the two forming daughter cells. Dur-ing cytokinesis, centralspindlin is truly central, as it organizes microtubule bundle struc-tures, recruits other factors to the site of division, and anchors the plasma membrane at the inter-cellular bridge while the two daughter cells are waiting for the final separation. Centralspindlin is a heterotetramer composed of two molecules of a kinesin-6 motor sub-unit, MKLP1, and two molecules of the second subunit, CYK4. For efficient microtubule Funding: We acknowledge support from the Cancer Research UK (programme grants C19769/A6356 and A11985 and equipment grant C19769/A7164 to MM), the Biotechnology and Biological Sciences Research Council (BBSRC, studentship to TD), the Nakajima Foundation (a research grant to NK), the MEXT Japan (Grant-in-Aid for Basic Research S, Project ID: 20221006 to TA), the Wellcome Trust and the Medical Research Council (MRC) joint funding (089703/Z/09/Z to CFK), the MRC (MR/K015850/1 to CFK), the Alzheimer's Research UK (ARUK-bundling, both the microtubule motor subunit MKLP1 and the non-motor CYK4 subunit are required. However, it has remained unclear how CYK4 contributes to this activity. Here, we took a combinatorial approach to investigate this process, using in vitro reconsti-tution and structural analyses by atomic force microscopy and Förster resonance energy transfer. We revealed that the CYK4 dimer binds to a hitherto unknown globular domain at the neck of the MKLP1 dimer and optimizes the configuration of two motor domains, making them suitable for antiparallel microtubule bundling. This provides novel insight into how other kinesin superfamily molecules with non-canonical neck structures may work.
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title = {CYK4 promotes antiparallel microtubule bundling by optimizing MKLP1 neck conformation},
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year = {2015},
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pages = {e1002121},
volume = {13},
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abstract = {Centralspindlin, a constitutive 2:2 heterotetramer of MKLP1 (a kinesin-6) and the non-motor subunit CYK4, plays important roles in cytokinesis. It is crucial for the formation of central spindle microtubule bundle structure. Its accumulation at the central antiparallel overlap zone is key for recruitment and regulation of downstream cytokinesis factors and for stable anchoring of the plasma membrane at the midbody. Both MKLP1 and CYK4 are required for efficient microtubule bundling. However, the mechanism by which CYK4 contributes to this is unclear. Here we performed structural and functional analyses of centralspindlin using high-speed atomic force microscopy, Förster resonance energy transfer analysis, and in vitro reconstitution. Our data reveal that CYK4 binds to a globular mass in the atypi-cally long MKLP1 neck domain between the catalytic core and the coiled coil and thereby reconfigures the two motor domains in the MKLP1 dimer to be suitable for antiparallel micro-tubule bundling. Our work provides insights into the microtubule bundling during cytokinesis and into the working mechanisms of the kinesins with non-canonical neck structures. Author Summary Cell division requires coordination of many different cellular components. Cytokinesis is the process by which the cytoplasm divides between the two forming daughter cells. Dur-ing cytokinesis, centralspindlin is truly central, as it organizes microtubule bundle struc-tures, recruits other factors to the site of division, and anchors the plasma membrane at the inter-cellular bridge while the two daughter cells are waiting for the final separation. Centralspindlin is a heterotetramer composed of two molecules of a kinesin-6 motor sub-unit, MKLP1, and two molecules of the second subunit, CYK4. For efficient microtubule Funding: We acknowledge support from the Cancer Research UK (programme grants C19769/A6356 and A11985 and equipment grant C19769/A7164 to MM), the Biotechnology and Biological Sciences Research Council (BBSRC, studentship to TD), the Nakajima Foundation (a research grant to NK), the MEXT Japan (Grant-in-Aid for Basic Research S, Project ID: 20221006 to TA), the Wellcome Trust and the Medical Research Council (MRC) joint funding (089703/Z/09/Z to CFK), the MRC (MR/K015850/1 to CFK), the Alzheimer's Research UK (ARUK-bundling, both the microtubule motor subunit MKLP1 and the non-motor CYK4 subunit are required. However, it has remained unclear how CYK4 contributes to this activity. Here, we took a combinatorial approach to investigate this process, using in vitro reconsti-tution and structural analyses by atomic force microscopy and Förster resonance energy transfer. We revealed that the CYK4 dimer binds to a hitherto unknown globular domain at the neck of the MKLP1 dimer and optimizes the configuration of two motor domains, making them suitable for antiparallel microtubule bundling. This provides novel insight into how other kinesin superfamily molecules with non-canonical neck structures may work.},
bibtype = {article},
author = {Davies, Tim and Kodera, Noriyuki and Kaminski Schierle, Gabriele S and Rees, Eric and Erdelyi, Miklos and Kaminski, Clemens F and Ando, Toshio and Mishima, Masanori and Gould, Kathleen L},
journal = {PLoS Biology},
number = {4}
}
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It is crucial for the formation of central spindle microtubule bundle structure. Its accumulation at the central antiparallel overlap zone is key for recruitment and regulation of downstream cytokinesis factors and for stable anchoring of the plasma membrane at the midbody. Both MKLP1 and CYK4 are required for efficient microtubule bundling. However, the mechanism by which CYK4 contributes to this is unclear. Here we performed structural and functional analyses of centralspindlin using high-speed atomic force microscopy, Förster resonance energy transfer analysis, and in vitro reconstitution. Our data reveal that CYK4 binds to a globular mass in the atypi-cally long MKLP1 neck domain between the catalytic core and the coiled coil and thereby reconfigures the two motor domains in the MKLP1 dimer to be suitable for antiparallel micro-tubule bundling. Our work provides insights into the microtubule bundling during cytokinesis and into the working mechanisms of the kinesins with non-canonical neck structures. Author Summary Cell division requires coordination of many different cellular components. Cytokinesis is the process by which the cytoplasm divides between the two forming daughter cells. Dur-ing cytokinesis, centralspindlin is truly central, as it organizes microtubule bundle struc-tures, recruits other factors to the site of division, and anchors the plasma membrane at the inter-cellular bridge while the two daughter cells are waiting for the final separation. Centralspindlin is a heterotetramer composed of two molecules of a kinesin-6 motor sub-unit, MKLP1, and two molecules of the second subunit, CYK4. For efficient microtubule Funding: We acknowledge support from the Cancer Research UK (programme grants C19769/A6356 and A11985 and equipment grant C19769/A7164 to MM), the Biotechnology and Biological Sciences Research Council (BBSRC, studentship to TD), the Nakajima Foundation (a research grant to NK), the MEXT Japan (Grant-in-Aid for Basic Research S, Project ID: 20221006 to TA), the Wellcome Trust and the Medical Research Council (MRC) joint funding (089703/Z/09/Z to CFK), the MRC (MR/K015850/1 to CFK), the Alzheimer's Research UK (ARUK-bundling, both the microtubule motor subunit MKLP1 and the non-motor CYK4 subunit are required. However, it has remained unclear how CYK4 contributes to this activity. Here, we took a combinatorial approach to investigate this process, using in vitro reconsti-tution and structural analyses by atomic force microscopy and Förster resonance energy transfer. We revealed that the CYK4 dimer binds to a hitherto unknown globular domain at the neck of the MKLP1 dimer and optimizes the configuration of two motor domains, making them suitable for antiparallel microtubule bundling. This provides novel insight into how other kinesin superfamily molecules with non-canonical neck structures may work.","bibtype":"article","author":"Davies, Tim and Kodera, Noriyuki and Kaminski Schierle, Gabriele S and Rees, Eric and Erdelyi, Miklos and Kaminski, Clemens F and Ando, Toshio and Mishima, Masanori and Gould, Kathleen L","journal":"PLoS Biology","number":"4","bibtex":"@article{\n title = {CYK4 promotes antiparallel microtubule bundling by optimizing MKLP1 neck conformation},\n type = {article},\n year = {2015},\n identifiers = {[object Object]},\n keywords = {biophysics,microscopy},\n pages = {e1002121},\n volume = {13},\n publisher = {Public Library of Science},\n id = {cfeb0144-a62d-318f-af80-931b8a5f217a},\n created = {2016-04-27T11:42:57.000Z},\n accessed = {2016-04-18},\n file_attached = {true},\n profile_id = {9b1c14de-b75a-3e23-a406-ef006d6fed26},\n last_modified = {2017-03-25T02:42:51.878Z},\n read = {false},\n starred = {false},\n authored = {true},\n confirmed = {true},\n hidden = {false},\n citation_key = {davies2015cyk4},\n source_type = {article},\n folder_uuids = {6e034f54-5623-437a-811f-80d304e37fd9},\n abstract = {Centralspindlin, a constitutive 2:2 heterotetramer of MKLP1 (a kinesin-6) and the non-motor subunit CYK4, plays important roles in cytokinesis. 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Our work provides insights into the microtubule bundling during cytokinesis and into the working mechanisms of the kinesins with non-canonical neck structures. Author Summary Cell division requires coordination of many different cellular components. Cytokinesis is the process by which the cytoplasm divides between the two forming daughter cells. Dur-ing cytokinesis, centralspindlin is truly central, as it organizes microtubule bundle struc-tures, recruits other factors to the site of division, and anchors the plasma membrane at the inter-cellular bridge while the two daughter cells are waiting for the final separation. Centralspindlin is a heterotetramer composed of two molecules of a kinesin-6 motor sub-unit, MKLP1, and two molecules of the second subunit, CYK4. 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