{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,5]],"date-time":"2026-03-05T22:16:01Z","timestamp":1772748961276,"version":"3.50.1"},"reference-count":43,"publisher":"Rockefeller University Press","issue":"5","content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2013,9,2]]},"abstract":"<jats:p>Spindle assembly checkpoint proteins have been thought to reside in the peripheral corona region of the kinetochore, distal to microtubule attachment sites at the outer plate. However, recent biochemical evidence indicates that checkpoint proteins are closely linked to the core kinetochore microtubule attachment site comprised of the Knl1\u2013Mis12\u2013Ndc80 (KMN) complexes\/KMN network. In this paper, we show that the Knl1\u2013Zwint1 complex is required to recruit the Rod\u2013Zwilch\u2013Zw10 (RZZ) and Mad1\u2013Mad2 complexes to the outer kinetochore. Consistent with this, nanometer-scale mapping indicates that RZZ, Mad1\u2013Mad2, and the C terminus of the dynein recruitment factor Spindly are closely juxtaposed with the KMN network in metaphase cells when their dissociation is blocked and the checkpoint is active. In contrast, the N terminus of Spindly is \u223c75 nm outside the calponin homology domain of the Ndc80 complex. These results reveal how checkpoint proteins are integrated within the substructure of the kinetochore and will aid in understanding the coordination of microtubule attachment and checkpoint signaling during chromosome segregation.<\/jats:p>","DOI":"10.1083\/jcb.201304197","type":"journal-article","created":{"date-parts":[[2013,8,27]],"date-time":"2013-08-27T03:41:22Z","timestamp":1377574882000},"page":"735-746","update-policy":"https:\/\/doi.org\/10.1083\/jcb.crossmarkpolicy","source":"Crossref","is-referenced-by-count":69,"title":["Spindle assembly checkpoint proteins are positioned close to core microtubule attachment sites at kinetochores"],"prefix":"10.1083","volume":"202","author":[{"given":"Dileep","family":"Varma","sequence":"first","affiliation":[{"name":"Department of Biology, The University of North Carolina-Chapel Hill, Chapel Hill, NC 27599 1"}]},{"given":"Xiaohu","family":"Wan","sequence":"additional","affiliation":[{"name":"Department of Biology, The University of North Carolina-Chapel Hill, Chapel Hill, NC 27599 1"}]},{"given":"Dhanya","family":"Cheerambathur","sequence":"additional","affiliation":[{"name":"Ludwig Institute for Cancer Research 2 and 3"},{"name":"Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093 2 and 3"},{"name":"Ludwig Institute for Cancer Research 2 and 3"},{"name":"Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093 2 and 3"}]},{"given":"Reto","family":"Gassmann","sequence":"additional","affiliation":[{"name":"Ludwig Institute for Cancer Research 2 and 3"},{"name":"Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093 2 and 3"},{"name":"Ludwig Institute for Cancer Research 2 and 3"},{"name":"Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093 2 and 3"}]},{"given":"Aussie","family":"Suzuki","sequence":"additional","affiliation":[{"name":"Department of Biology, The University of North Carolina-Chapel Hill, Chapel Hill, NC 27599 1"}]},{"given":"Josh","family":"Lawrimore","sequence":"additional","affiliation":[{"name":"Department of Biology, The University of North Carolina-Chapel Hill, Chapel Hill, NC 27599 1"}]},{"given":"Arshad","family":"Desai","sequence":"additional","affiliation":[{"name":"Ludwig Institute for Cancer Research 2 and 3"},{"name":"Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093 2 and 3"},{"name":"Ludwig Institute for Cancer Research 2 and 3"},{"name":"Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093 2 and 3"}]},{"given":"E.D.","family":"Salmon","sequence":"additional","affiliation":[{"name":"Department of Biology, The University of North Carolina-Chapel Hill, Chapel Hill, NC 27599 1"}]}],"member":"291","published-online":{"date-parts":[[2013,8,26]]},"reference":[{"key":"2023072306033388800_bib1","doi-asserted-by":"publisher","first-page":"1968","DOI":"10.1091\/mbc.E09-04-0356","article-title":"Spindly\/CCDC99 is required for efficient chromosome congression and mitotic checkpoint regulation","volume":"21","author":"Barisic","year":"2010","journal-title":"Mol. Biol. Cell."},{"key":"2023072306033388800_bib2","doi-asserted-by":"publisher","first-page":"56","DOI":"10.1016\/j.cub.2003.12.025","article-title":"In vivo dynamics of the rough deal checkpoint protein during Drosophila mitosis","volume":"14","author":"Basto","year":"2004","journal-title":"Curr. Biol."},{"key":"2023072306033388800_bib3","doi-asserted-by":"publisher","first-page":"856","DOI":"10.1016\/j.cub.2005.03.052","article-title":"Recruitment of Mad2 to the kinetochore requires the Rod\/Zw10 complex","volume":"15","author":"Buffin","year":"2005","journal-title":"Curr. Biol."},{"key":"2023072306033388800_bib4","doi-asserted-by":"publisher","first-page":"944","DOI":"10.1038\/35046598","article-title":"Human Zw10 and ROD are mitotic checkpoint proteins that bind to kinetochores","volume":"2","author":"Chan","year":"2000","journal-title":"Nat. Cell Biol."},{"key":"2023072306033388800_bib5","doi-asserted-by":"publisher","first-page":"859","DOI":"10.1083\/jcb.200812167","article-title":"Mitotic control of kinetochore-associated dynein and spindle orientation by human Spindly","volume":"185","author":"Chan","year":"2009","journal-title":"J. Cell Biol."},{"key":"2023072306033388800_bib6","doi-asserted-by":"publisher","first-page":"587","DOI":"10.1091\/mbc.E07-10-1051","article-title":"KNL1 and the CENP-H\/I\/K complex coordinately direct kinetochore assembly in vertebrates","volume":"19","author":"Cheeseman","year":"2008","journal-title":"Mol. Biol. Cell."},{"key":"2023072306033388800_bib7","doi-asserted-by":"publisher","first-page":"616","DOI":"10.1016\/j.str.2010.02.014","article-title":"Structural analysis of the RZZ complex reveals common ancestry with multisubunit vesicle tethering machinery","volume":"18","author":"Civril","year":"2010","journal-title":"Structure."},{"key":"2023072306033388800_bib8","doi-asserted-by":"publisher","first-page":"2103","DOI":"10.1016\/j.cub.2003.10.056","article-title":"Nuf2 and Hec1 are required for retention of the checkpoint proteins Mad1 and Mad2 to kinetochores","volume":"13","author":"DeLuca","year":"2003","journal-title":"Curr. Biol."},{"key":"2023072306033388800_bib9","doi-asserted-by":"publisher","first-page":"519","DOI":"10.1091\/mbc.E04-09-0852","article-title":"Hec1 and nuf2 are core components of the kinetochore outer plate essential for organizing microtubule attachment sites","volume":"16","author":"DeLuca","year":"2005","journal-title":"Mol. Biol. Cell."},{"key":"2023072306033388800_bib10","doi-asserted-by":"publisher","first-page":"2143","DOI":"10.1016\/j.cub.2007.11.037","article-title":"Aurora B kinase-dependent recruitment of hZW10 and hROD to tensionless kinetochores","volume":"17","author":"Famulski","year":"2007","journal-title":"Curr. Biol."},{"key":"2023072306033388800_bib11","doi-asserted-by":"publisher","first-page":"507","DOI":"10.1083\/jcb.200708021","article-title":"Stable hZW10 kinetochore residency, mediated by hZwint-1 interaction, is essential for the mitotic checkpoint","volume":"180","author":"Famulski","year":"2008","journal-title":"J. Cell Biol."},{"key":"2023072306033388800_bib12","doi-asserted-by":"publisher","first-page":"2385","DOI":"10.1101\/gad.1687508","article-title":"A new mechanism controlling kinetochore-microtubule interactions revealed by comparison of two dynein-targeting components: SPDL-1 and the Rod\/Zwilch\/Zw10 complex","volume":"22","author":"Gassmann","year":"2008","journal-title":"Genes Dev."},{"key":"2023072306033388800_bib13","doi-asserted-by":"publisher","first-page":"957","DOI":"10.1101\/gad.1886810","article-title":"Removal of Spindly from microtubule-attached kinetochores controls spindle checkpoint silencing in human cells","volume":"24","author":"Gassmann","year":"2010","journal-title":"Genes Dev."},{"key":"2023072306033388800_bib14","doi-asserted-by":"publisher","first-page":"1005","DOI":"10.1083\/jcb.200702062","article-title":"Spindly, a novel protein essential for silencing the spindle assembly checkpoint, recruits dynein to the kinetochore","volume":"177","author":"Griffis","year":"2007","journal-title":"J. Cell Biol."},{"key":"2023072306033388800_bib15","doi-asserted-by":"publisher","first-page":"1778","DOI":"10.1016\/j.cub.2008.08.012","article-title":"Kinetochore-microtubule attachment relies on the disordered N-terminal tail domain of Hec1","volume":"18","author":"Guimaraes","year":"2008","journal-title":"Curr. Biol."},{"key":"2023072306033388800_bib16","doi-asserted-by":"publisher","first-page":"1995","DOI":"10.1091\/mbc.12.7.1995","article-title":"Microtubule-dependent changes in assembly of microtubule motor proteins and mitotic spindle checkpoint proteins at PtK1 kinetochores","volume":"12","author":"Hoffman","year":"2001","journal-title":"Mol. Biol. Cell."},{"key":"2023072306033388800_bib17","doi-asserted-by":"publisher","first-page":"1159","DOI":"10.1083\/jcb.200105093","article-title":"Cytoplasmic dynein\/dynactin drives kinetochore protein transport to the spindle poles and has a role in mitotic spindle checkpoint inactivation","volume":"155","author":"Howell","year":"2001","journal-title":"J. Cell Biol."},{"key":"2023072306033388800_bib18","doi-asserted-by":"publisher","first-page":"953","DOI":"10.1016\/j.cub.2004.05.053","article-title":"Spindle checkpoint protein dynamics at kinetochores in living cells","volume":"14","author":"Howell","year":"2004","journal-title":"Curr. Biol."},{"key":"2023072306033388800_bib19","doi-asserted-by":"publisher","first-page":"386","DOI":"10.1016\/j.tcb.2005.05.003","article-title":"Rod-Zw10-Zwilch: a key player in the spindle checkpoint","volume":"15","author":"Karess","year":"2005","journal-title":"Trends Cell Biol."},{"key":"2023072306033388800_bib20","doi-asserted-by":"publisher","first-page":"3318","DOI":"10.1091\/mbc.E11-03-0213","article-title":"Zwint-1 is a novel Aurora B substrate required for the assembly of a dynein-binding platform on kinetochores","volume":"22","author":"Kasuboski","year":"2011","journal-title":"Mol. Biol. Cell."},{"key":"2023072306033388800_bib21","doi-asserted-by":"publisher","first-page":"6549","DOI":"10.1073\/pnas.1118210109","article-title":"Structure of human Mad1 C-terminal domain reveals its involvement in kinetochore targeting","volume":"109","author":"Kim","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072306033388800_bib22","doi-asserted-by":"publisher","first-page":"739","DOI":"10.1083\/jcb.151.4.739","article-title":"Dynein is a transient kinetochore component whose binding is regulated by microtubule attachment, not tension","volume":"151","author":"King","year":"2000","journal-title":"J. Cell Biol."},{"key":"2023072306033388800_bib23","doi-asserted-by":"publisher","first-page":"663","DOI":"10.1016\/j.devcel.2007.09.005","article-title":"Human Blinkin\/AF15q14 is required for chromosome alignment and the mitotic checkpoint through direct interaction with Bub1 and BubR1","volume":"13","author":"Kiyomitsu","year":"2007","journal-title":"Dev. Cell."},{"key":"2023072306033388800_bib24","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1083\/jcb.200411118","article-title":"ZW10 links mitotic checkpoint signaling to the structural kinetochore","volume":"169","author":"Kops","year":"2005","journal-title":"J. Cell Biol."},{"key":"2023072306033388800_bib25","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1016\/j.tcb.2010.10.007","article-title":"Sensing centromere tension: Aurora B and the regulation of kinetochore function","volume":"21","author":"Lampson","year":"2011","journal-title":"Trends Cell Biol."},{"key":"2023072306033388800_bib26","doi-asserted-by":"publisher","first-page":"R966","DOI":"10.1016\/j.cub.2012.10.006","article-title":"The spindle assembly checkpoint","volume":"22","author":"Lara-Gonzalez","year":"2012","journal-title":"Curr. Biol."},{"key":"2023072306033388800_bib27","doi-asserted-by":"publisher","first-page":"6901","DOI":"10.1038\/sj.onc.1209687","article-title":"Hec1 sequentially recruits Zwint-1 and ZW10 to kinetochores for faithful chromosome segregation and spindle checkpoint control","volume":"25","author":"Lin","year":"2006","journal-title":"Oncogene."},{"key":"2023072306033388800_bib28","doi-asserted-by":"publisher","first-page":"597","DOI":"10.1016\/S0076-6879(03)60130-8","article-title":"Spinning disk confocal microscope system for rapid high-resolution, multimode, fluorescence speckle microscopy and green fluorescent protein imaging in living cells","volume":"360","author":"Maddox","year":"2003","journal-title":"Methods Enzymol."},{"key":"2023072306033388800_bib29","doi-asserted-by":"publisher","first-page":"475","DOI":"10.1038\/ncb2223","article-title":"Constitutive Mad1 targeting to kinetochores uncouples checkpoint signalling from chromosome biorientation","volume":"13","author":"Maldonado","year":"2011","journal-title":"Nat. Cell Biol."},{"key":"2023072306033388800_bib30","doi-asserted-by":"publisher","first-page":"825","DOI":"10.1242\/jcs.064790","article-title":"Welcome to a new kind of tension: translating kinetochore mechanics into a wait-anaphase signal","volume":"123","author":"Maresca","year":"2010","journal-title":"J. Cell Sci."},{"key":"2023072306033388800_bib31","doi-asserted-by":"publisher","first-page":"2163","DOI":"10.1007\/s00018-010-0322-x","article-title":"Contrasting models for kinetochore microtubule attachment in mammalian cells","volume":"67","author":"McEwen","year":"2010","journal-title":"Cell. Mol. Life Sci."},{"key":"2023072306033388800_bib32","doi-asserted-by":"publisher","first-page":"3595","DOI":"10.1098\/rstb.2011.0072","article-title":"Spindle assembly checkpoint: the third decade","volume":"366","author":"Musacchio","year":"2011","journal-title":"Philos. Trans. R. Soc. Lond. B Biol. Sci."},{"key":"2023072306033388800_bib33","doi-asserted-by":"publisher","first-page":"379","DOI":"10.1038\/nrm2163","article-title":"The spindle-assembly checkpoint in space and time","volume":"8","author":"Musacchio","year":"2007","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"2023072306033388800_bib34","doi-asserted-by":"publisher","first-page":"e7640","DOI":"10.1371\/journal.pone.0007640","article-title":"Roles for the conserved spc105p\/kre28p complex in kinetochore-microtubule binding and the spindle assembly checkpoint","volume":"4","author":"Pagliuca","year":"2009","journal-title":"PLoS ONE."},{"key":"2023072306033388800_bib35","doi-asserted-by":"publisher","first-page":"835","DOI":"10.1083\/jcb.201002070","article-title":"The MIS12 complex is a protein interaction hub for outer kinetochore assembly","volume":"190","author":"Petrovic","year":"2010","journal-title":"J. Cell Biol."},{"key":"2023072306033388800_bib36","doi-asserted-by":"crossref","first-page":"1939","DOI":"10.1242\/jcs.113.11.1939","article-title":"HZwint-1, a novel human kinetochore component that interacts with HZW10","volume":"113","author":"Starr","year":"2000","journal-title":"J. Cell Sci."},{"key":"2023072306033388800_bib37","doi-asserted-by":"publisher","first-page":"759","DOI":"10.1091\/mbc.E10-08-0671","article-title":"The NDC80 complex proteins Nuf2 and Hec1 make distinct contributions to kinetochore-microtubule attachment in mitosis","volume":"22","author":"Sundin","year":"2011","journal-title":"Mol. Biol. Cell."},{"key":"2023072306033388800_bib38","doi-asserted-by":"publisher","first-page":"5927","DOI":"10.1242\/jcs.093724","article-title":"The KMN protein network\u2014chief conductors of the kinetochore orchestra","volume":"125","author":"Varma","year":"2012","journal-title":"J. Cell Sci."},{"key":"2023072306033388800_bib39","doi-asserted-by":"publisher","first-page":"672","DOI":"10.1016\/j.cell.2009.03.035","article-title":"Protein architecture of the human kinetochore microtubule attachment site","volume":"137","author":"Wan","year":"2009","journal-title":"Cell."},{"key":"2023072306033388800_bib40","doi-asserted-by":"publisher","first-page":"1035","DOI":"10.1091\/mbc.E11-09-0767","article-title":"The coupling between sister kinetochore directional instability and oscillations in centromere stretch in metaphase PtK1 cells","volume":"23","author":"Wan","year":"2012","journal-title":"Mol. Biol. Cell."},{"key":"2023072306033388800_bib41","doi-asserted-by":"publisher","first-page":"54590","DOI":"10.1074\/jbc.M407588200","article-title":"Human Zwint-1 specifies localization of Zeste White 10 to kinetochores and is essential for mitotic checkpoint signaling","volume":"279","author":"Wang","year":"2004","journal-title":"J. Biol. Chem."},{"key":"2023072306033388800_bib42","doi-asserted-by":"publisher","first-page":"1001","DOI":"10.1038\/ncb1101-1001","article-title":"Kinetochore dynein: its dynamics and role in the transport of the Rough deal checkpoint protein","volume":"3","author":"Wojcik","year":"2001","journal-title":"Nat. Cell Biol."},{"key":"2023072306033388800_bib43","doi-asserted-by":"publisher","first-page":"187","DOI":"10.1083\/jcb.200805185","article-title":"SPDL-1 functions as a kinetochore receptor for MDF-1 in Caenorhabditis elegans","volume":"183","author":"Yamamoto","year":"2008","journal-title":"J. Cell Biol."}],"container-title":["Journal of Cell Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/rupress.org\/jcb\/article-pdf\/202\/5\/735\/1580634\/jcb_201304197.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/rupress.org\/jcb\/article-pdf\/202\/5\/735\/1580634\/jcb_201304197.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,23]],"date-time":"2023-07-23T06:05:40Z","timestamp":1690092340000},"score":1,"resource":{"primary":{"URL":"https:\/\/rupress.org\/jcb\/article\/202\/5\/735\/37202\/Spindle-assembly-checkpoint-proteins-are"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,8,26]]},"references-count":43,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2013,9,2]]}},"URL":"https:\/\/doi.org\/10.1083\/jcb.201304197","relation":{},"ISSN":["1540-8140","0021-9525"],"issn-type":[{"value":"1540-8140","type":"electronic"},{"value":"0021-9525","type":"print"}],"subject":[],"published":{"date-parts":[[2013,8,26]]}}}