Protein Architecture of the Human Kinetochore Microtubule Attachment Site
Chromosome segregation requires assembly of kinetochores on centromeric chromatin to mediate interactions with spindle microtubules and control cell-cycle progression. To elucidate the protein architecture of human kinetochores, we developed a two-color fluorescence light microscopy method that meas...
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| Veröffentlicht in: | Cell 2009-05, Vol.137 (4), p.672-684 |
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| creator | Wan, Xiaohu O'Quinn, Ryan P. Pierce, Heather L. Joglekar, Ajit P. Gall, Walt E. DeLuca, Jennifer G. Carroll, Christopher W. Liu, Song-Tao Yen, Tim J. McEwen, Bruce F. Stukenberg, P. Todd Desai, Arshad Salmon, E.D. |
| description | Chromosome segregation requires assembly of kinetochores on centromeric chromatin to mediate interactions with spindle microtubules and control cell-cycle progression. To elucidate the protein architecture of human kinetochores, we developed a two-color fluorescence light microscopy method that measures average label separation, Delta, at |
| doi_str_mv | 10.1016/j.cell.2009.03.035 |
| format | Article |
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Todd ; Desai, Arshad ; Salmon, E.D.</creator><creatorcontrib>Wan, Xiaohu ; O'Quinn, Ryan P. ; Pierce, Heather L. ; Joglekar, Ajit P. ; Gall, Walt E. ; DeLuca, Jennifer G. ; Carroll, Christopher W. ; Liu, Song-Tao ; Yen, Tim J. ; McEwen, Bruce F. ; Stukenberg, P. Todd ; Desai, Arshad ; Salmon, E.D.</creatorcontrib><description>Chromosome segregation requires assembly of kinetochores on centromeric chromatin to mediate interactions with spindle microtubules and control cell-cycle progression. To elucidate the protein architecture of human kinetochores, we developed a two-color fluorescence light microscopy method that measures average label separation, Delta, at <5 nm accuracy. Delta analysis of 16 proteins representing core structural complexes spanning the centromeric chromatin-microtubule interface, when correlated with mechanical states of spindle-attached kinetochores, provided a nanometer-scale map of protein position and mechanical properties of protein linkages. Treatment with taxol, which suppresses microtubule dynamics and activates the spindle checkpoint, revealed a specific switch in kinetochore architecture. Cumulatively, Delta analysis revealed that compliant linkages are restricted to the proximity of chromatin, suggested a model for how the KMN (KNL1/Mis12 complex/Ndc80 complex) network provides microtubule attachment and generates pulling forces from depolymerization, and identified an intrakinetochore molecular switch that may function in controlling checkpoint activity.</description><identifier>ISSN: 0092-8674</identifier><identifier>EISSN: 1097-4172</identifier><identifier>DOI: 10.1016/j.cell.2009.03.035</identifier><identifier>PMID: 19450515</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>CELLCYLE ; Cytoskeletal Proteins ; DNA-Binding Proteins - metabolism ; HeLa Cells ; Humans ; Kinetochores - chemistry ; Kinetochores - metabolism ; Metaphase ; Microscopy, Fluorescence ; Microtubule-Associated Proteins - metabolism ; Microtubules - chemistry ; Microtubules - metabolism ; Nuclear Proteins ; PROTEINS</subject><ispartof>Cell, 2009-05, Vol.137 (4), p.672-684</ispartof><rights>2009 Elsevier Inc.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c519t-88996dd35f1f3fe6520df368f85253f4db1c6230199959f8371929fd76c1ced23</citedby><cites>FETCH-LOGICAL-c519t-88996dd35f1f3fe6520df368f85253f4db1c6230199959f8371929fd76c1ced23</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.cell.2009.03.035$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>230,315,781,785,886,3551,27929,27930,46000</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19450515$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wan, Xiaohu</creatorcontrib><creatorcontrib>O'Quinn, Ryan P.</creatorcontrib><creatorcontrib>Pierce, Heather L.</creatorcontrib><creatorcontrib>Joglekar, Ajit P.</creatorcontrib><creatorcontrib>Gall, Walt E.</creatorcontrib><creatorcontrib>DeLuca, Jennifer G.</creatorcontrib><creatorcontrib>Carroll, Christopher W.</creatorcontrib><creatorcontrib>Liu, Song-Tao</creatorcontrib><creatorcontrib>Yen, Tim J.</creatorcontrib><creatorcontrib>McEwen, Bruce F.</creatorcontrib><creatorcontrib>Stukenberg, P. Todd</creatorcontrib><creatorcontrib>Desai, Arshad</creatorcontrib><creatorcontrib>Salmon, E.D.</creatorcontrib><title>Protein Architecture of the Human Kinetochore Microtubule Attachment Site</title><title>Cell</title><addtitle>Cell</addtitle><description>Chromosome segregation requires assembly of kinetochores on centromeric chromatin to mediate interactions with spindle microtubules and control cell-cycle progression. To elucidate the protein architecture of human kinetochores, we developed a two-color fluorescence light microscopy method that measures average label separation, Delta, at <5 nm accuracy. Delta analysis of 16 proteins representing core structural complexes spanning the centromeric chromatin-microtubule interface, when correlated with mechanical states of spindle-attached kinetochores, provided a nanometer-scale map of protein position and mechanical properties of protein linkages. Treatment with taxol, which suppresses microtubule dynamics and activates the spindle checkpoint, revealed a specific switch in kinetochore architecture. Cumulatively, Delta analysis revealed that compliant linkages are restricted to the proximity of chromatin, suggested a model for how the KMN (KNL1/Mis12 complex/Ndc80 complex) network provides microtubule attachment and generates pulling forces from depolymerization, and identified an intrakinetochore molecular switch that may function in controlling checkpoint activity.</description><subject>CELLCYLE</subject><subject>Cytoskeletal Proteins</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>HeLa Cells</subject><subject>Humans</subject><subject>Kinetochores - chemistry</subject><subject>Kinetochores - metabolism</subject><subject>Metaphase</subject><subject>Microscopy, Fluorescence</subject><subject>Microtubule-Associated Proteins - metabolism</subject><subject>Microtubules - chemistry</subject><subject>Microtubules - metabolism</subject><subject>Nuclear Proteins</subject><subject>PROTEINS</subject><issn>0092-8674</issn><issn>1097-4172</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9UUtL7DAUDqLo-PgDLqQrd52bR5M2IMIgvriKgroOnfTEZmgbTVLBf2_KDF7vRjiQxffK-Q5CxwTPCSbiz2quoevmFGM5xywN30IzgmWZF6Sk22iWAJpXoiz20H4IK4xxxTnfRXtEFhxzwmfo9tG7CHbIFl63NoKOo4fMmSy2kN2MfT1kf-0A0enWJeDe6sQfl2MH2SLGWrc9DDF7SspDtGPqLsDR5j1AL1eXzxc3-d3D9e3F4i7XnMiYV5WUomkYN8QwA4JT3BgmKlNxypkpmiXRgjJMpJRcmoqVRFJpmlJooqGh7ACdr33fxmUPjU75vu7Um7d97T-Vq636Hxlsq17dh6JCSsxxMjjdGHj3PkKIqrdharIewI1BiZIWZVlMSXRNTDuH4MF8hxCspguolZp0arqAwiwNT6KTn9_7J9lUnghnawKkkj4seBW0hSHtZn2qXzXO_ub_BeGVmG0</recordid><startdate>20090515</startdate><enddate>20090515</enddate><creator>Wan, Xiaohu</creator><creator>O'Quinn, Ryan P.</creator><creator>Pierce, Heather L.</creator><creator>Joglekar, Ajit P.</creator><creator>Gall, Walt E.</creator><creator>DeLuca, Jennifer G.</creator><creator>Carroll, Christopher W.</creator><creator>Liu, Song-Tao</creator><creator>Yen, Tim J.</creator><creator>McEwen, Bruce F.</creator><creator>Stukenberg, P. Todd</creator><creator>Desai, Arshad</creator><creator>Salmon, E.D.</creator><general>Elsevier Inc</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20090515</creationdate><title>Protein Architecture of the Human Kinetochore Microtubule Attachment Site</title><author>Wan, Xiaohu ; O'Quinn, Ryan P. ; Pierce, Heather L. ; Joglekar, Ajit P. ; Gall, Walt E. ; DeLuca, Jennifer G. ; Carroll, Christopher W. ; Liu, Song-Tao ; Yen, Tim J. ; McEwen, Bruce F. ; Stukenberg, P. 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Todd</creatorcontrib><creatorcontrib>Desai, Arshad</creatorcontrib><creatorcontrib>Salmon, E.D.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Cell</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wan, Xiaohu</au><au>O'Quinn, Ryan P.</au><au>Pierce, Heather L.</au><au>Joglekar, Ajit P.</au><au>Gall, Walt E.</au><au>DeLuca, Jennifer G.</au><au>Carroll, Christopher W.</au><au>Liu, Song-Tao</au><au>Yen, Tim J.</au><au>McEwen, Bruce F.</au><au>Stukenberg, P. Todd</au><au>Desai, Arshad</au><au>Salmon, E.D.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Protein Architecture of the Human Kinetochore Microtubule Attachment Site</atitle><jtitle>Cell</jtitle><addtitle>Cell</addtitle><date>2009-05-15</date><risdate>2009</risdate><volume>137</volume><issue>4</issue><spage>672</spage><epage>684</epage><pages>672-684</pages><issn>0092-8674</issn><eissn>1097-4172</eissn><abstract>Chromosome segregation requires assembly of kinetochores on centromeric chromatin to mediate interactions with spindle microtubules and control cell-cycle progression. To elucidate the protein architecture of human kinetochores, we developed a two-color fluorescence light microscopy method that measures average label separation, Delta, at <5 nm accuracy. Delta analysis of 16 proteins representing core structural complexes spanning the centromeric chromatin-microtubule interface, when correlated with mechanical states of spindle-attached kinetochores, provided a nanometer-scale map of protein position and mechanical properties of protein linkages. Treatment with taxol, which suppresses microtubule dynamics and activates the spindle checkpoint, revealed a specific switch in kinetochore architecture. 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| subjects | CELLCYLE Cytoskeletal Proteins DNA-Binding Proteins - metabolism HeLa Cells Humans Kinetochores - chemistry Kinetochores - metabolism Metaphase Microscopy, Fluorescence Microtubule-Associated Proteins - metabolism Microtubules - chemistry Microtubules - metabolism Nuclear Proteins PROTEINS |
| title | Protein Architecture of the Human Kinetochore Microtubule Attachment Site |
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