CytoskeletonPub Date : 2024-09-02DOI: 10.1002/cm.21923
Jashaswi Basu, Chaitanya A. Athale
{"title":"Collective effect of Vigna sp. (mung) tubulin GTP hydrolysis rate divergence on microtubule filament assembly","authors":"Jashaswi Basu, Chaitanya A. Athale","doi":"10.1002/cm.21923","DOIUrl":"10.1002/cm.21923","url":null,"abstract":"<p>Microtubules (MTs) are dynamic cytoskeletal filaments with highly conserved sequences across evolution, polymerizing by the GTP-dependent assembly of tubulin subunits. Despite the sequence conservation, MT polymerization kinetics diverge quantitatively between vertebrate brain, the model plant <i>Arabidopsis</i> and the protozoan <i>Plasmodium</i>. Previously, tubulin purified from seedlings of the plant <i>Vigna</i> sp. (mung) by temperature cycling was found to have a very low critical concentration. However, the lengths of MTs were sub-micron, much shorter than brain tubulin filaments. This was explained in simulations to be the result of the collective effect of high nucleation and GTP hydrolysis rates. Here, we test the effect of GTPase rates of affinity-purified <i>Vigna</i> sp. tubulin on microtubule polymerization and elongation. Affinity-purified mung tubulin is active and has a critical concentration of .37 μM. The GTP-dependent polymerization kinetics are transient, consistent with previous results. Polymerization is stabilized in the presence of either GTP analog GMPPNP (non-hydrolyzable) or GMPCPP (slow-hydrolyzable). Using interference reflection microscopy (IRM) we find polymerization with the non-hydrolysable analog significantly increases filament numbers, while lengths are unaffected for both GTP analogs. However, prolonged incubation with slow-hydrolyzable GMPCPP results in long filaments, pointing to GTP hydrolysis as a key factor determining MT length. We find the average GTPase turnover number of mung tubulin is 22.8 min<sup>−1</sup>, compared to 2.04 min<sup>−1</sup> for goat brain tubulin. Thus modulating GTPase rates affects both nucleation and elongation. This quantitative divergence in kinetics despite high sequence conservation in the GTPase domains of <i>α</i>- and <i>β</i>-tubulin could help better understand the roles of selective pressure and function in the diverse organisms.</p>","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"82 4","pages":"242-259"},"PeriodicalIF":2.4,"publicationDate":"2024-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142115588","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
CytoskeletonPub Date : 2024-08-26DOI: 10.1002/cm.21920
Albin E. Berg
{"title":"Author profile: Albin E. Berg","authors":"Albin E. Berg","doi":"10.1002/cm.21920","DOIUrl":"10.1002/cm.21920","url":null,"abstract":"","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"81 12","pages":"917-918"},"PeriodicalIF":2.4,"publicationDate":"2024-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142057481","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
CytoskeletonPub Date : 2024-08-26DOI: 10.1002/cm.21917
Brooke E. Waechtler
{"title":"Author profile: Brooke E. Waechtler","authors":"Brooke E. Waechtler","doi":"10.1002/cm.21917","DOIUrl":"10.1002/cm.21917","url":null,"abstract":"","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"81 12","pages":"915-916"},"PeriodicalIF":2.4,"publicationDate":"2024-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142057482","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
CytoskeletonPub Date : 2024-08-18DOI: 10.1002/cm.21906
{"title":"Front Cover Image","authors":"","doi":"10.1002/cm.21906","DOIUrl":"https://doi.org/10.1002/cm.21906","url":null,"abstract":"<p>ON THE FRONT COVER: Nematic organization of growing actin filaments generated using agent-based simulations.</p><p>Credit: Deb Sankar Banerjee (Carnegie Mellon University; The University of Chicago) and Shiladitya Banerjee (Carnegie Mellon University)\u0000\u0000 <figure>\u0000 <div><picture>\u0000 <source></source></picture><p></p>\u0000 </div>\u0000 </figure>\u0000 </p>","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"81 8","pages":"C1"},"PeriodicalIF":2.4,"publicationDate":"2024-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cm.21906","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142002524","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
CytoskeletonPub Date : 2024-08-18DOI: 10.1002/cm.21908
{"title":"Inner Back Cover Image","authors":"","doi":"10.1002/cm.21908","DOIUrl":"https://doi.org/10.1002/cm.21908","url":null,"abstract":"<p>ON THE INNER BACK COVER: Short and finite microtubule tactoids in presence of anti-parallel crosslinker, MAP65. In absence of salt, microtubules form short tactoids like formations.</p><p>Credit: Prashali Chauhan and Jennifer Ross (Syracuse University)\u0000\u0000 <figure>\u0000 <div><picture>\u0000 <source></source></picture><p></p>\u0000 </div>\u0000 </figure>\u0000 </p>","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"81 8","pages":"C3"},"PeriodicalIF":2.4,"publicationDate":"2024-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cm.21908","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142002599","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
CytoskeletonPub Date : 2024-08-18DOI: 10.1002/cm.21909
{"title":"Back Cover Image","authors":"","doi":"10.1002/cm.21909","DOIUrl":"https://doi.org/10.1002/cm.21909","url":null,"abstract":"<p>ON THE BACK COVER: Fasin-bundled actin networks encapsulated inside giant unilamellar vesicles</p><p>Credit: Nadab H. Wubshet (University of Michigan; Harvard University) and Allen Liu (University of Michigan)\u0000\u0000 <figure>\u0000 <div><picture>\u0000 <source></source></picture><p></p>\u0000 </div>\u0000 </figure>\u0000 </p>","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"81 8","pages":"C4"},"PeriodicalIF":2.4,"publicationDate":"2024-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cm.21909","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142002617","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
CytoskeletonPub Date : 2024-08-18DOI: 10.1002/cm.21907
{"title":"Inner Front Cover Image","authors":"","doi":"10.1002/cm.21907","DOIUrl":"https://doi.org/10.1002/cm.21907","url":null,"abstract":"<p>ON THE INNER FRONT COVER: Long microtubule bundles. As we increase the salt, we observe long microtubule bundles.</p><p>Credit: Prashali Chauhan and Jennifer Ross (Syracuse University)\u0000\u0000 <figure>\u0000 <div><picture>\u0000 <source></source></picture><p></p>\u0000 </div>\u0000 </figure>\u0000 </p>","PeriodicalId":55186,"journal":{"name":"Cytoskeleton","volume":"81 8","pages":"C2"},"PeriodicalIF":2.4,"publicationDate":"2024-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cm.21907","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142002590","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}