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An isolated CLASP TOG domain suppresses microtubule catastrophe and promotes rescue

Microtubules are heavily regulated dynamic polymers of αβ-tubulin that are required for proper chromosome segregation and organization of the cytoplasm. Polymerases in the XMAP215 family use arrayed TOG domains to promote faster microtubule elongation. Regulatory factors in the cytoplasmic linker as...

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Autores principales: Majumdar, Shreoshi, Kim, Tae, Chen, Zhe, Munyoki, Sarah, Tso, Shih-Chia, Brautigam, Chad A., Rice, Luke M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5994897/
https://www.ncbi.nlm.nih.gov/pubmed/29851564
http://dx.doi.org/10.1091/mbc.E17-12-0748
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author Majumdar, Shreoshi
Kim, Tae
Chen, Zhe
Munyoki, Sarah
Tso, Shih-Chia
Brautigam, Chad A.
Rice, Luke M.
author_facet Majumdar, Shreoshi
Kim, Tae
Chen, Zhe
Munyoki, Sarah
Tso, Shih-Chia
Brautigam, Chad A.
Rice, Luke M.
author_sort Majumdar, Shreoshi
collection PubMed
description Microtubules are heavily regulated dynamic polymers of αβ-tubulin that are required for proper chromosome segregation and organization of the cytoplasm. Polymerases in the XMAP215 family use arrayed TOG domains to promote faster microtubule elongation. Regulatory factors in the cytoplasmic linker associated protein (CLASP) family that reduce catastrophe and/or increase rescue also contain arrayed TOGs, but how CLASP TOGs contribute to activity is poorly understood. Here, using Saccharomyces cerevisiae Stu1 as a model CLASP, we report structural, biochemical, and reconstitution studies that clarify functional properties of CLASP TOGs. The two TOGs in Stu1 have very different tubulin-binding properties: TOG2 binds to both unpolymerized and polymerized tubulin, and TOG1 binds very weakly to either. The structure of Stu1-TOG2 reveals a CLASP-specific residue that likely confers distinctive tubulin-binding properties. The isolated TOG2 domain strongly suppresses microtubule catastrophe and increases microtubule rescue in vitro, contradicting the expectation that regulatory activity requires an array of TOGs. Single point mutations on the tubulin-binding surface of TOG2 ablate its anti-catastrophe and rescue activity in vitro, and Stu1 function in cells. Revealing that an isolated CLASP TOG can regulate polymerization dynamics without being part of an array provides insight into the mechanism of CLASPs and diversifies the understanding of TOG function.
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spelling pubmed-59948972018-08-16 An isolated CLASP TOG domain suppresses microtubule catastrophe and promotes rescue Majumdar, Shreoshi Kim, Tae Chen, Zhe Munyoki, Sarah Tso, Shih-Chia Brautigam, Chad A. Rice, Luke M. Mol Biol Cell Articles Microtubules are heavily regulated dynamic polymers of αβ-tubulin that are required for proper chromosome segregation and organization of the cytoplasm. Polymerases in the XMAP215 family use arrayed TOG domains to promote faster microtubule elongation. Regulatory factors in the cytoplasmic linker associated protein (CLASP) family that reduce catastrophe and/or increase rescue also contain arrayed TOGs, but how CLASP TOGs contribute to activity is poorly understood. Here, using Saccharomyces cerevisiae Stu1 as a model CLASP, we report structural, biochemical, and reconstitution studies that clarify functional properties of CLASP TOGs. The two TOGs in Stu1 have very different tubulin-binding properties: TOG2 binds to both unpolymerized and polymerized tubulin, and TOG1 binds very weakly to either. The structure of Stu1-TOG2 reveals a CLASP-specific residue that likely confers distinctive tubulin-binding properties. The isolated TOG2 domain strongly suppresses microtubule catastrophe and increases microtubule rescue in vitro, contradicting the expectation that regulatory activity requires an array of TOGs. Single point mutations on the tubulin-binding surface of TOG2 ablate its anti-catastrophe and rescue activity in vitro, and Stu1 function in cells. Revealing that an isolated CLASP TOG can regulate polymerization dynamics without being part of an array provides insight into the mechanism of CLASPs and diversifies the understanding of TOG function. The American Society for Cell Biology 2018-06-01 /pmc/articles/PMC5994897/ /pubmed/29851564 http://dx.doi.org/10.1091/mbc.E17-12-0748 Text en © 2018 Majumdar et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. http://creativecommons.org/licenses/by-nc-sa/3.0/ This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License.
spellingShingle Articles
Majumdar, Shreoshi
Kim, Tae
Chen, Zhe
Munyoki, Sarah
Tso, Shih-Chia
Brautigam, Chad A.
Rice, Luke M.
An isolated CLASP TOG domain suppresses microtubule catastrophe and promotes rescue
title An isolated CLASP TOG domain suppresses microtubule catastrophe and promotes rescue
title_full An isolated CLASP TOG domain suppresses microtubule catastrophe and promotes rescue
title_fullStr An isolated CLASP TOG domain suppresses microtubule catastrophe and promotes rescue
title_full_unstemmed An isolated CLASP TOG domain suppresses microtubule catastrophe and promotes rescue
title_short An isolated CLASP TOG domain suppresses microtubule catastrophe and promotes rescue
title_sort isolated clasp tog domain suppresses microtubule catastrophe and promotes rescue
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5994897/
https://www.ncbi.nlm.nih.gov/pubmed/29851564
http://dx.doi.org/10.1091/mbc.E17-12-0748
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