Regulation of Kif15 localization and motility by the C-terminus of TPX2 and microtubule dynamics

Mann, Barbara J.; Balchand, Sai Keshavan; Wadsworth, Patricia

doi:10.1091/mbc.e16-06-0476

Cited by 27 publications

(48 citation statements)

References 52 publications

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“…However, in metazoan, kinesin-12 displays microtubule bundling and sliding activity. Recently, formation of tetrameric HsKif15 was reported also in vivo [53]. In vitro, dimeric HsKif15 promotes microtubule sliding of higher order microtubules utilizing a motile motor in concert with a non-motor microtubule binding domain [50].…”

Section: Pok2 Acts Redundantly With Pok1 In Division Site Maintenancementioning

confidence: 95%

“…Other in vitro studies report tetrameric HsKif15 switching between microtubule tracks and HsKif15 motor collectives that display processive movement at steady velocity [52]. Recently, formation of tetrameric HsKif15 was reported also in vivo [53]. Whether POK2 molecules form di-or tetramers and how these interact with microtubules requires further scrutiny.…”

Section: Pok2 Acts Redundantly With Pok1 In Division Site Maintenancementioning

confidence: 99%

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Dual localized kinesin‐12 POK 2 plays multiple roles during cell division and interacts with MAP 65‐3

et al. 2018

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Kinesins are versatile nano-machines that utilize variable non-motor domains to tune specific motor microtubule encounters. During plant cytokinesis, the kinesin-12 orthologs, PHRAGMOPLAST ORIENTING KINESIN (POK)1 and POK2, are essential for rapid centrifugal expansion of the cytokinetic apparatus, the phragmoplast, toward a pre-selected cell plate fusion site at the cell cortex. Here, we report on the spatio-temporal localization pattern of POK2, mediated by distinct protein domains. Functional dissection of POK2 domains revealed the association of POK2 with the site of the future cell division plane and with the phragmoplast during cytokinesis. Accumulation of POK2 at the phragmoplast midzone depends on its functional POK2 motor domain and is fine-tuned by its carboxy-terminal region that also directs POK2 to the division site. Furthermore, POK2 likely stabilizes the phragmoplast midzone via interaction with the conserved microtubule-associated protein MAP65-3/PLEIADE, a well-established microtubule cross-linker. Collectively, our results suggest that dual localized POK2 plays multiple roles during plant cell division.

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Section: Pok2 Acts Redundantly With Pok1 In Division Site Maintenancementioning

confidence: 95%

Section: Pok2 Acts Redundantly With Pok1 In Division Site Maintenancementioning

confidence: 99%

Dual localized kinesin‐12 POK 2 plays multiple roles during cell division and interacts with MAP 65‐3

et al. 2018

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“…The previously controversial question of the oligomerisation state of Kif15 under physiological conditions now seems settled, as motors within mitotic or interphase extracts were confirmed to be tetrameric by single molecule imaging (Mann, Balchand et al 2017). However it remains to be established whether the functional dimer-of-dimers of Kif15 is stabilised by parallel or antiparallel interactions, or indeed a mixture of the two, as in the Drosophila kinesin-5 tetramer (Scholey, Nithianantham We take this to mean that the functional unit in our optical trapping experiments is the twin heads of a tethered dimer.…”

Section: The Mechanics Of Kif15 Are Distinct From Those Of Kinesin-1 mentioning

confidence: 99%

“…Several cell biological studies have suggested that the functional redundancy of Kif15 with Eg5 might reflect a similar stepping mechanism and a similar action on microtubules (Tanenbaum, Macurek et al 2009, Vanneste, Takagi et al 2009). By contrast, recent in vitro reconstitution work on the action of single Kif15 tetramers on microtubules has revealed Kif15-specific behaviours (Drechsler, McHugh et al 2014, Drechsler and McAinsh 2016, Mann, Balchand et al 2017. Unlike Eg5 (Valentine, Fordyce et al 2006), Kif15 motors at zero load are highly processive, being capable of moving over 1μm along a microtubule before detaching.…”

Section: Introductionmentioning

confidence: 97%

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Single molecule mechanics reveal Kif15 as an active molecular ratchet with acute strain sensitivity

McHugh

Drechsler

McAinsh

et al. 2017

Preprint

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Human Kif15 is a tetrameric kinesin-12 that contributes critically to bipolar spindle assembly in eukaryotes. Here we examine its single molecule mechanics. Under hindering loads, Kif15 steps predominantly towards microtubule plus ends, with its forestep:backstep ratio decreasing exponentially with load and stall occurring at ~6pN. Between steps, Kif15 binds stably, usually via a single head domain. By complete contrast, under assisting loads, Kif15 detaches rapidly, even in AMPPNP. Furthermore, Kif15 can autoinhibit, via an interaction requiring its C-terminus. Autoinhibited Kif15 binds microtubules nucleotide-independently, resists both hindering and assisting loads, and is further stabilized by Tpx2, which interacts with the Kif15 C-terminus. Our data reveal the mechanics of Kif15 to be extraordinarily sensitive to loading direction. When unloaded, it walks rapidly; when pulled forwards it slips and when pulled backwards it grips. We discuss the implications of this unique mechanical behaviour for the roles of Kif15 in spindle function.

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Phragmoplast Orienting Kinesin 2 Is a Weak Motor Switching between Processive and Diffusive Modes

Chugh

Reißner

Bugiel

et al. 2018

Biophysical Journal

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Plant development and morphology relies on the accurate insertion of new cell walls during cytokinesis. However, how a plant cell correctly orients a new wall is poorly understood. Two kinesin class-12 members, phragmoplast orienting kinesin 1 (POK1) and POK2, are involved in the process, but how these molecular machines work is not known. Here, we used in vivo and single-molecule in vitro measurements to determine how Arabidopsis thaliana POK2 motors function mechanically. We found that POK2 is a very weak, on average plus-end-directed, moderately fast kinesin. Interestingly, POK2 switches between processive and diffusive modes characterized by an exclusive-state mean-squared-displacement analysis. Our results support a model that POK motors push against peripheral microtubules of the phragmoplast for its guidance. This pushing model may mechanically explain the conspicuous narrowing of the division site. Together, our findings provide mechanical insight into how active motors accurately position new cell walls in plants.

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Regulation of Kif15 localization and motility by the C-terminus of TPX2 and microtubule dynamics

Abstract: Temporal and spatial regulation of mitotic motor proteins is required for proper spindle assembly and function. A single microtubule-associated protein, TPX2, regulates both kinesin-5 and kinesin-12 family members to support bipolar spindle formation and maintenance.

Cited by 27 publications

References 52 publications

Dual localized kinesin‐12 POK 2 plays multiple roles during cell division and interacts with MAP 65‐3

Dual localized kinesin‐12 POK 2 plays multiple roles during cell division and interacts with MAP 65‐3

Single molecule mechanics reveal Kif15 as an active molecular ratchet with acute strain sensitivity

Phragmoplast Orienting Kinesin 2 Is a Weak Motor Switching between Processive and Diffusive Modes

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