Functional overlap of microtubule assembly factors in chromatin-promoted spindle assembly
Distinct pathways from centrosomes and chromatin are thought to contribute in parallel to microtubule nucleation and stabilization during animal cell mitotic spindle assembly, but their full mechanisms are not known. We investigated the function of three proposed nucleation/stabilization factors, TP...
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| Veröffentlicht in: | Molecular biology of the cell 2009-06, Vol.20 (11), p.2766-2773 |
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| creator | Groen, Aaron C Maresca, Thomas J Gatlin, Jesse C Salmon, Edward D Mitchison, Timothy J |
| description | Distinct pathways from centrosomes and chromatin are thought to contribute in parallel to microtubule nucleation and stabilization during animal cell mitotic spindle assembly, but their full mechanisms are not known. We investigated the function of three proposed nucleation/stabilization factors, TPX2, gamma-tubulin and XMAP215, in chromatin-promoted assembly of anastral spindles in Xenopus laevis egg extract. In addition to conventional depletion-add back experiments, we tested whether factors could substitute for each other, indicative of functional redundancy. All three factors were required for microtubule polymerization and bipolar spindle assembly around chromatin beads. Depletion of TPX2 was partially rescued by the addition of excess XMAP215 or EB1, or inhibiting MCAK (a Kinesin-13). Depletion of either gamma-tubulin or XMAP215 was partially rescued by adding back XMAP215, but not by adding any of the other factors. These data reveal functional redundancy between specific assembly factors in the chromatin pathway, suggesting individual proteins or pathways commonly viewed to be essential may not have entirely unique functions. |
| doi_str_mv | 10.1091/mbc.E09-01-0043 |
| format | Article |
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We investigated the function of three proposed nucleation/stabilization factors, TPX2, gamma-tubulin and XMAP215, in chromatin-promoted assembly of anastral spindles in Xenopus laevis egg extract. In addition to conventional depletion-add back experiments, we tested whether factors could substitute for each other, indicative of functional redundancy. All three factors were required for microtubule polymerization and bipolar spindle assembly around chromatin beads. Depletion of TPX2 was partially rescued by the addition of excess XMAP215 or EB1, or inhibiting MCAK (a Kinesin-13). Depletion of either gamma-tubulin or XMAP215 was partially rescued by adding back XMAP215, but not by adding any of the other factors. These data reveal functional redundancy between specific assembly factors in the chromatin pathway, suggesting individual proteins or pathways commonly viewed to be essential may not have entirely unique functions.</description><identifier>ISSN: 1059-1524</identifier><identifier>EISSN: 1939-4586</identifier><identifier>DOI: 10.1091/mbc.E09-01-0043</identifier><identifier>PMID: 19369413</identifier><language>eng</language><publisher>United States: The American Society for Cell Biology</publisher><subject>Animals ; Cell Cycle Proteins - genetics ; Cell Cycle Proteins - metabolism ; Chromatin - metabolism ; Female ; Green Fluorescent Proteins - genetics ; Green Fluorescent Proteins - metabolism ; Humans ; Kinesin - metabolism ; Kinetics ; Microscopy, Fluorescence - methods ; Microtubule-Associated Proteins - genetics ; Microtubule-Associated Proteins - metabolism ; Microtubules - metabolism ; Nuclear Proteins - genetics ; Nuclear Proteins - metabolism ; Oocytes - cytology ; Oocytes - metabolism ; Phosphoproteins - genetics ; Phosphoproteins - metabolism ; Spindle Apparatus - metabolism ; Time Factors ; Tubulin - genetics ; Tubulin - metabolism ; Xenopus laevis ; Xenopus Proteins - genetics ; Xenopus Proteins - metabolism</subject><ispartof>Molecular biology of the cell, 2009-06, Vol.20 (11), p.2766-2773</ispartof><rights>2009 by The American Society for Cell Biology 2009</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c469t-9aef1bab49995070869eeb1d7e879e39970563edd6c9ca3baad4b855cc8e6f863</citedby><cites>FETCH-LOGICAL-c469t-9aef1bab49995070869eeb1d7e879e39970563edd6c9ca3baad4b855cc8e6f863</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2688555/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2688555/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,723,776,780,881,27903,27904,53769,53771</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19369413$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Weis, Karsten</contributor><creatorcontrib>Groen, Aaron C</creatorcontrib><creatorcontrib>Maresca, Thomas J</creatorcontrib><creatorcontrib>Gatlin, Jesse C</creatorcontrib><creatorcontrib>Salmon, Edward D</creatorcontrib><creatorcontrib>Mitchison, Timothy J</creatorcontrib><title>Functional overlap of microtubule assembly factors in chromatin-promoted spindle assembly</title><title>Molecular biology of the cell</title><addtitle>Mol Biol Cell</addtitle><description>Distinct pathways from centrosomes and chromatin are thought to contribute in parallel to microtubule nucleation and stabilization during animal cell mitotic spindle assembly, but their full mechanisms are not known. 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These data reveal functional redundancy between specific assembly factors in the chromatin pathway, suggesting individual proteins or pathways commonly viewed to be essential may not have entirely unique functions.</description><subject>Animals</subject><subject>Cell Cycle Proteins - genetics</subject><subject>Cell Cycle Proteins - metabolism</subject><subject>Chromatin - metabolism</subject><subject>Female</subject><subject>Green Fluorescent Proteins - genetics</subject><subject>Green Fluorescent Proteins - metabolism</subject><subject>Humans</subject><subject>Kinesin - metabolism</subject><subject>Kinetics</subject><subject>Microscopy, Fluorescence - methods</subject><subject>Microtubule-Associated Proteins - genetics</subject><subject>Microtubule-Associated Proteins - metabolism</subject><subject>Microtubules - metabolism</subject><subject>Nuclear Proteins - genetics</subject><subject>Nuclear Proteins - metabolism</subject><subject>Oocytes - cytology</subject><subject>Oocytes - metabolism</subject><subject>Phosphoproteins - genetics</subject><subject>Phosphoproteins - metabolism</subject><subject>Spindle Apparatus - metabolism</subject><subject>Time Factors</subject><subject>Tubulin - genetics</subject><subject>Tubulin - metabolism</subject><subject>Xenopus laevis</subject><subject>Xenopus Proteins - genetics</subject><subject>Xenopus Proteins - metabolism</subject><issn>1059-1524</issn><issn>1939-4586</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkctLxDAQxoMouj7O3qQnb12TTZp2LoLI-gDBix48hSSdupG2qUm74H9vll18nGZgfvPN8H2EnDM6ZxTYVWfsfEkhpyynVPA9MmPAIRdFJfdTTwvIWbEQR-Q4xg9KmRCyPCRHCZIgGJ-Rt7upt6PzvW4zv8bQ6iHzTdY5G_w4manFTMeInWm_skbb0YeYuT6zq-A7Pbo-H1LjR6yzOLi-_oOfkoNGtxHPdvWEvN4tX24f8qfn-8fbm6fcCgljDhobZrQRAFDQklYSEA2rS6xKQA5Q0kJyrGtpwWputK6FqYrC2gplU0l-Qq63usNkOqwt9mPQrRqC63T4Ul479X_Su5V692u1kFXSKZLA5U4g-M8J46g6Fy22re7RT1HJkjNOF1UCr7Zg8ibGgM3PEUbVJg6V4lBIQVGmNnGkjYu_v_3yO__5N8oWipY</recordid><startdate>200906</startdate><enddate>200906</enddate><creator>Groen, Aaron C</creator><creator>Maresca, Thomas J</creator><creator>Gatlin, Jesse C</creator><creator>Salmon, Edward D</creator><creator>Mitchison, Timothy J</creator><general>The American Society for Cell Biology</general><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>200906</creationdate><title>Functional overlap of microtubule assembly factors in chromatin-promoted spindle assembly</title><author>Groen, Aaron C ; Maresca, Thomas J ; Gatlin, Jesse C ; Salmon, Edward D ; Mitchison, Timothy J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c469t-9aef1bab49995070869eeb1d7e879e39970563edd6c9ca3baad4b855cc8e6f863</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Animals</topic><topic>Cell Cycle Proteins - genetics</topic><topic>Cell Cycle Proteins - metabolism</topic><topic>Chromatin - metabolism</topic><topic>Female</topic><topic>Green Fluorescent Proteins - genetics</topic><topic>Green Fluorescent Proteins - metabolism</topic><topic>Humans</topic><topic>Kinesin - metabolism</topic><topic>Kinetics</topic><topic>Microscopy, Fluorescence - methods</topic><topic>Microtubule-Associated Proteins - genetics</topic><topic>Microtubule-Associated Proteins - metabolism</topic><topic>Microtubules - metabolism</topic><topic>Nuclear Proteins - genetics</topic><topic>Nuclear Proteins - metabolism</topic><topic>Oocytes - cytology</topic><topic>Oocytes - metabolism</topic><topic>Phosphoproteins - genetics</topic><topic>Phosphoproteins - metabolism</topic><topic>Spindle Apparatus - metabolism</topic><topic>Time Factors</topic><topic>Tubulin - genetics</topic><topic>Tubulin - metabolism</topic><topic>Xenopus laevis</topic><topic>Xenopus Proteins - genetics</topic><topic>Xenopus Proteins - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Groen, Aaron C</creatorcontrib><creatorcontrib>Maresca, Thomas J</creatorcontrib><creatorcontrib>Gatlin, Jesse C</creatorcontrib><creatorcontrib>Salmon, Edward D</creatorcontrib><creatorcontrib>Mitchison, Timothy J</creatorcontrib><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>Molecular biology of the cell</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Groen, Aaron C</au><au>Maresca, Thomas J</au><au>Gatlin, Jesse C</au><au>Salmon, Edward D</au><au>Mitchison, Timothy J</au><au>Weis, Karsten</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Functional overlap of microtubule assembly factors in chromatin-promoted spindle assembly</atitle><jtitle>Molecular biology of the cell</jtitle><addtitle>Mol Biol Cell</addtitle><date>2009-06</date><risdate>2009</risdate><volume>20</volume><issue>11</issue><spage>2766</spage><epage>2773</epage><pages>2766-2773</pages><issn>1059-1524</issn><eissn>1939-4586</eissn><abstract>Distinct pathways from centrosomes and chromatin are thought to contribute in parallel to microtubule nucleation and stabilization during animal cell mitotic spindle assembly, but their full mechanisms are not known. 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| subjects | Animals Cell Cycle Proteins - genetics Cell Cycle Proteins - metabolism Chromatin - metabolism Female Green Fluorescent Proteins - genetics Green Fluorescent Proteins - metabolism Humans Kinesin - metabolism Kinetics Microscopy, Fluorescence - methods Microtubule-Associated Proteins - genetics Microtubule-Associated Proteins - metabolism Microtubules - metabolism Nuclear Proteins - genetics Nuclear Proteins - metabolism Oocytes - cytology Oocytes - metabolism Phosphoproteins - genetics Phosphoproteins - metabolism Spindle Apparatus - metabolism Time Factors Tubulin - genetics Tubulin - metabolism Xenopus laevis Xenopus Proteins - genetics Xenopus Proteins - metabolism |
| title | Functional overlap of microtubule assembly factors in chromatin-promoted spindle assembly |
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