The kinesin-like ncd protein of Drosophila is a minus end-directed microtubule motor

McDonald, Heather B.; Stewart, Russell J.; Goldstein, Lawrence S.B.

doi:10.1016/0092-8674(90)90412-8

Cited by 329 publications

(227 citation statements)

References 24 publications

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Section: Resultssupporting

confidence: 77%

“…The rate of MT movement calculated by dividing the width of the bleached zone by the time of the linear increase of the fluorescence in the bleached zone was 0.105 Ϯ 0.001 m/s. The numbers obtained by two independent experimental approaches were consistent with each other and with the velocity of Ncd-based MT movement in vitro determined using a MT gliding assay (0.06 -0.16 m/s (McDonald et al, 1990;Walker et al, 1990;deCastro et al, 1999;Tao et al, 2006). These results strongly suggest that MT movement seen in GFP-Ncd-overexpressing cells is indeed generated by the motor activity of Ncd.…”

Section: Resultssupporting

confidence: 71%

“…Many of the filaments that incorporated fluorescently tagged tubulin were significantly brighter than others, as would be expected if they were composed of several MTs (Figure 2, right). Therefore, the labeling pattern of MTs in Ncd-expressing cells is consistent with the results of in vitro experiments (McDonald et al, 1990;Chandra et al, 1993) that indicate that Ncd causes the formation of MT bundles.To test directly whether Ncd-dependent MT cross-linking results in movement of MTs against each other, we examined the behavior of MTs in cells overexpressing GFP-Ncd. Live imaging of MT behavior in Ncd-transfected cells showed that in such cells, the shapes of MT cables constantly changed.…”

supporting

confidence: 77%

“…These motors have two MT binding sites, an ATP-dependent site in the motor domain and an ATP-independent site in the tail, and they have been shown to induce MT bundling (McDonald et al, 1990;Chandra et al, 1993;Karabay and Walker, 1999;Matuliene et al, 1999). Recent small interfering RNA (siRNA) studies demonstrate that knockdown of the kinesin-14 family member Ncd results in the formation of splayed mitotic spindle poles, suggesting that the activity of this motor is required for focusing of the MTs at the pole (Goshima and Vale, 2003;Morales-Mulia and Scholey, 2005).…”

Section: Introductionmentioning

confidence: 99%

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Microtubule Motor Ncd Induces Sliding of Microtubules In Vivo

Oladipo

Cowan

Rodionov

2007

MBoC

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The mitotic spindle is a microtubule (MT)-based molecular machine that serves for equal segregation of chromosomes during cell division. The formation of the mitotic spindle requires the activity of MT motors, including members of the kinesin-14 family. Although evidence suggests that kinesins-14 act by driving the sliding of MT bundles in different areas of the spindle, such sliding activity had never been demonstrated directly. To test the hypothesis that kinesins-14 can induce MT sliding in living cells, we developed an in vivo assay, which involves overexpression of the kinesin-14 family member Drosophila Ncd in interphase mammalian fibroblasts. We found that green fluorescent protein (GFP)-Ncd colocalized with cytoplasmic MTs, whose distribution was determined by microinjection of Cy3 tubulin into GFPtransfected cells. Ncd overexpression resulted in the formation of MT bundles that exhibited dynamic "looping" behavior never observed in control cells. Photobleaching studies and fluorescence speckle microscopy analysis demonstrated that neighboring MTs in bundles could slide against each other with velocities of 0.1 m/s, corresponding to the velocities of movement of the recombinant Ncd in in vitro motility assays. Our data, for the first time, demonstrate generation of sliding forces between adjacent MTs by Ncd, and they confirm the proposed roles of kinesins-14 in the mitotic spindle morphogenesis. INTRODUCTIONThe mitotic spindle is a molecular machine that serves for equal segregation of chromosomes during mitosis. The principal structural components of the mitotic spindle are cytoplasmic microtubules (MTs) organized into two polarized (minus ends at the center) radial arrays, located at a distance from each other. MT organization in the mitotic spindle is achieved by MT motors, kinesins and dyneins, which generate force for movement toward the plus or minus ends of MTs and that participate in the transport of chromosomes to the spindle poles, spindle elongation during anaphase, regulation of the MT turnover rates, and spindle morphogenesis (for review, see Sharp et al., 2000b;Karsenti and Vernos, 2001;McIntosh et al., 2002;Scholey et al., 2003;Gadde and Heald, 2004;Wadsworth and Khodjakov, 2004).A special role in the formation and maintenance of the mitotic spindle belongs to the minus-end-directed members of the kinesin-14 subfamily. Unlike conventional kinesin, kinesin-14 family members have motor domains at the carboxy terminus (for review, see Ovechkina and Wordeman, 2003). The best studied kinesin-14 members are Drosophila Ncd (Komma et al., 1991;Matthies et al., 1996) and Saccharomyces cerevisiae Kar3 (Meluh and Rose, 1990). These MT motors, and their mammalian (Matuliene et al., 1999;Mountain et al., 1999;Zhu et al., 2005) and plant (Vanstraelen et al., 2006) homologues, play essential roles in mitosis and meiosis. Their activities are important for the formation of the mitotic spindle poles (Goshima and Vale, 2003; MoralesMulia and Scholey, 2005;Zhu et al., 2005) and for regulation of the distance...

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Section: Resultssupporting

confidence: 77%

Section: Resultssupporting

confidence: 71%

supporting

confidence: 77%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Microtubule Motor Ncd Induces Sliding of Microtubules In Vivo

Oladipo

Cowan

Rodionov

2007

MBoC

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“…The amino terminal part of KCBP (2.5 N fusion protein) containing the globular tail and coiled-coil stalk region but lacking the motor domain did not show clear binding to tubulin subunits. Some KLPs, especially minus end motors, have been shown to contain a second microtubule binding site in the tail region that is insensitive to ATP (Chandra etal., 1993;Endow etaL, 1994;Kuriyama etaL, 1995;Liao et al, 1994;McDonald et al, 1990). Although KCBP aminoterminal fusion protein does not appear to interact with tubulin subunits, we can not'rule out the possibility that it may interact with microtubules.…”

Section: Discussionmentioning

confidence: 89%