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Two Kinesin-14A Motors Oligomerize to Drive Poleward Microtubule Convergence for Acentrosomal Spindle Morphogenesis in Arabidopsis thaliana

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dc.contributor.author Hotta, Takashi en
dc.contributor.author Lee, Yuh-Ru Julie en
dc.contributor.author Higaki, Takumi en
dc.contributor.author Hashimoto, Takashi en
dc.contributor.author Liu, Bo en
dc.date.accessioned 2022-07-25T04:08:31Z en
dc.date.available 2022-07-25T04:08:31Z en
dc.date.issued 2022-07-22 en
dc.identifier.uri http://hdl.handle.net/10061/14761 en
dc.description.abstract Plant cells form acentrosomal spindles with microtubules (MTs) converged toward two structurally undefined poles by employing MT minus end-directed Kinesin-14 motors. To date, it is unclear whether the convergent bipolar MT array assumes unified poles in plant spindles, and if so, how such a goal is achieved. Among six classes of Kinesin-14 motors in Arabidopsis thaliana, the Kinesin-14A motors ATK1 (KatA) and ATK5 share the essential function in spindle morphogenesis. To understand how the two functionally redundant Kinesin-14A motors contributed to the spindle assembly, we had ATK1-GFP and ATK5-GFP fusion proteins expressed in their corresponding null mutants and found that they were functionally comparable to their native forms. Although ATK1 was a nuclear protein and ATK5 cytoplasmic prior to nuclear envelop breakdown, at later mitotic stages, the two motors shared similar localization patterns of uniform association with both spindle and phragmoplast MTs. We found that ATK1 and ATK5 were rapidly concentrated toward unified polar foci when cells were under hyperosmotic conditions. Concomitantly, spindle poles became perfectly focused as if there were centrosome-like MT-organizing centers where ATK1 and ATK5 were highly enriched and at which kinetochore fibers pointed. The separation of ATK1/ATK5-highlighted MTs from those of kinetochore fibers suggested that the motors translocated interpolar MTs. Our protein purification and live-cell imaging results showed that ATK1 and ATK5 are associated with each other in vivo. The stress-induced spindle pole convergence was also accompanied by poleward accumulation of the MT nucleator γ-tubulin. These results led to the conclusion that the two Kinesin-14A motors formed oligomeric motor complexes that drove MT translocation toward the spindle pole to establish acentrosomal spindles with convergent poles. ja
dc.language.iso en en
dc.publisher Frontiers Media en
dc.relation.isreplacedby https://www.frontiersin.org/articles/10.3389/fcell.2022.949345/full en
dc.rights © 2022 Hotta, Lee, Higaki, Hashimoto and Liu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. ja
dc.subject kinesin-14 en
dc.subject kinetochore fibers en
dc.subject interpolar microtubules en
dc.subject microtubule convergence en
dc.subject mitotic spindle en
dc.subject spindle poles en
dc.subject Arabidopsis en
dc.title Two Kinesin-14A Motors Oligomerize to Drive Poleward Microtubule Convergence for Acentrosomal Spindle Morphogenesis in Arabidopsis thaliana en
dc.type.nii Journal Article en
dc.contributor.transcription ハシモト, タカシ ja
dc.contributor.alternative 橋本, 隆 ja
dc.textversion none en
dc.identifier.eissn 2296-634X en
dc.identifier.jtitle Frontiers in Cell and Developmental Biology en
dc.identifier.volume 10 en
dc.relation.doi 10.3389/fcell.2022.949345 en
dc.identifier.NAIST-ID 73290157 en

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