Binding of the CYK-4 Subunit of the Centralspindlin Complex Induces a Large Scale Conformational Change in the Kinesin Subunit

White, Erin A.; Raghuraman, H.; Perozo, Eduardo; Glotzer, Michael

doi:10.1074/jbc.m113.463695

Cited by 20 publications

(32 citation statements)

References 43 publications

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“…As homo-FRET reflects the intra-molecular geometry of the two Venus moieties, this indicates that C120 binding changes the relative positioning of the two Venus moieties in a dimer. This is consistent with a report by electron paramagnetic resonance [ 24 ] and confirms that the effect of CYK-4 on the configuration of the ZEN-4 motor domains observed by HS-AFM also occurs in solution. Because the ZEN-4 construct used for this analysis did not contain the motor domain, we conclude that CYK-4 changes the structure of the neck domain of the ZEN-4 dimer.…”

Section: Resultssupporting

confidence: 93%

“…Because of these unusual mechanistic features, the molecular structure of the heterotetrameric centralspindlin complex of CYK4 and MKLP1 is of great interest. Although a recent study using electron paramagnetic resonance spectroscopy reported a change in the mobility of ZEN-4 neck residues upon CYK-4 binding [ 24 ], it remains unclear how this influences the configuration of the two motor domains and how this modulates microtubule bundling by the complex.…”

Section: Introductionmentioning

confidence: 99%

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CYK4 Promotes Antiparallel Microtubule Bundling by Optimizing MKLP1 Neck Conformation

et al. 2015

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Centralspindlin, a constitutive 2:2 heterotetramer of MKLP1 (a kinesin-6) and the non-motor subunit CYK4, plays important roles in cytokinesis. It is crucial for the formation of central spindle microtubule bundle structure. Its accumulation at the central antiparallel overlap zone is key for recruitment and regulation of downstream cytokinesis factors and for stable anchoring of the plasma membrane at the midbody. Both MKLP1 and CYK4 are required for efficient microtubule bundling. However, the mechanism by which CYK4 contributes to this is unclear. Here we performed structural and functional analyses of centralspindlin using high-speed atomic force microscopy, Fӧrster resonance energy transfer analysis, and in vitro reconstitution. Our data reveal that CYK4 binds to a globular mass in the atypically long MKLP1 neck domain between the catalytic core and the coiled coil and thereby reconfigures the two motor domains in the MKLP1 dimer to be suitable for antiparallel microtubule bundling. Our work provides insights into the microtubule bundling during cytokinesis and into the working mechanisms of the kinesins with non-canonical neck structures.

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Section: Resultssupporting

confidence: 93%

Section: Introductionmentioning

confidence: 99%

CYK4 Promotes Antiparallel Microtubule Bundling by Optimizing MKLP1 Neck Conformation

et al. 2015

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“…Since previous studies had shown that Tum works as a molecular switch which activates the kinesin motor function of Pav (Davies et al, 2015; Tao et al, 2016; White et al, 2013), we examined whether the presence of Tum protein could affect the F-actin bundling and/or crosslinking activities of Pav. Bacterially-purified Tum protein alone does not bundle either F-actin or MTs (Fig.…”

Section: Resultsmentioning

confidence: 99%

The kinesin-like protein Pavarotti functions non-canonically to regulate actin dynamics

Nakamura

Verboon

Prentiss

et al. 2019

Preprint

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36Pavarotti, the Drosophila MKLP1 ortholog, is a kinesin-like protein that with Tumbleweed 37 (MgcRacGAP) works together as the centralspindlin complex. This complex is essential for 38 cytokinesis where it helps to organize the contractile actomyosin ring at the equator of dividing 39 cells by activating the RhoGEF Pebble. Actomyosin rings also function as the driving force 40 during cell wound repair. We previously showed that Tumbleweed and Pebble are required for 41 the cell wound repair process. Here, we show that Pavarotti also functions during wound repair 42 and confirm that while Pavarotti, Tumbleweed, and Pebble are utilized during this cellular repair, 43 it is not as the conserved centralspindlin complex. Surprisingly, in vitro and in vivo work show 44 that the classically microtubule-associated Pavarotti binds directly to actin and has a non-45 canonical role directly regulating actin dynamics. We show that Pavarotti also works 46 independently from Tumbleweed in several actin-related processes during the normal 47 48 49 Nakamura et al.

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“…Our results do not rule out the possibility that a partner protein could bind to KIF7’s extended neck linker to alter the effective length and/or structure in a manner that changes KIF7’s microtubule-based functions. This possibility was suggested by Davies et al [68] based on a) a comparison of kinesin motors with extended neck linkers and b) the finding that the extended neck linker of the kinesin-6 CeZen-4 serves as the binding site for the partner protein CeCyk-4 which alters Ce Zen-4 structure to slow motor velocity and enable microtubule bundling [68, 69]. Further support for this idea comes from work on Ce Zen-4’s homolog Dm Pav where binding of the Dm Tum partner enables multi-motor microtubule gliding activity [70].…”

Section: Discussionmentioning

confidence: 99%

Microtubule binding of the kinesin-4 KIF7 and its regulation by autoinhibition

Blasius

Yue

Verhey

2019

Preprint

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KIF7 is a member of the kinesin-4 family and plays critical roles in Hedgehog signaling in vertebrate cells. KIF7 is an atypical kinesin as it binds to microtubules but is immotile. We demonstrate that, like conventional kinesins, KIF7 is regulated by autoinhibition as the full-length motor cannot bind to microtubules whereas truncated versions bind statically to microtubules in cells. Previous work suggested that truncated KIF7 motors bind preferentially to the plus ends of microtubules in vitro, however, we find that truncated KIF7 does not bind preferentially to or track the plus ends of growing microtubules in mammalian cells or in cell extracts. Although the truncated KIF7 did alter microtubule dynamics in cells, this property is not specific to KIF7 as expression of an active kinesin-1 motor also altered microtubule growth rates. The immotile behavior of KIF7 is not due to the extended neck linker domain as its deletion does not activate KIF7 for motility and its presence in a KIF5C/KIF7 chimeric motor does not prevent processive motility. Together this work indicates that the atypical kinesin KIF7 is regulated by autoinhibition to prevent binding to microtubules and alteration of microtubule dynamics in cells.

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Binding of the CYK-4 Subunit of the Centralspindlin Complex Induces a Large Scale Conformational Change in the Kinesin Subunit

Cited by 20 publications

References 43 publications

CYK4 Promotes Antiparallel Microtubule Bundling by Optimizing MKLP1 Neck Conformation

CYK4 Promotes Antiparallel Microtubule Bundling by Optimizing MKLP1 Neck Conformation

The kinesin-like protein Pavarotti functions non-canonically to regulate actin dynamics

Microtubule binding of the kinesin-4 KIF7 and its regulation by autoinhibition

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