A Complex of NuMA and Cytoplasmic Dynein Is Essential for Mitotic Spindle Assembly

Merdes, Andreas; Ramyar, Kasra X.; Vechio, Janet D.; Cleveland, Don W.

doi:10.1016/s0092-8674(00)81365-3

Cited by 554 publications

(559 citation statements)

References 43 publications

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“…NuMA is required to tether centrosomes to spindle poles and to organize microtubule minus ends at spindle poles in vertebrate cells (Gaglio et al, 1995;Merdes et al, 1996;Gordon et al, 2001). Surprisingly, we show that perturbation of Kid function permits centrosomes to remain tethered to the spindle and microtubule minus ends to remain focused at spindle poles despite the perturbation of NuMA function.…”

Section: Spindle Organizationmentioning

confidence: 72%

“…However, several observations support the view that antibody injection completely and specifically inhibited the activities of both Kid and NuMA. First, defects in chromosome alignment and spindle morphology generated by injection of antibodies to either Kid or NuMA are equivalent to the effects observed upon the complete depletion of either of these proteins from extracts prepared from frog eggs (Merdes et al, 1996;Antonio et al, 2000;Funabiki and Murray, 2000). Second, the relationship between Kid and NuMA is specific because simultaneous injection of antibodies to Kid and either HSET or CENP-E showed no significant differences from injection of each antibody alone.…”

Section: Discussionmentioning

confidence: 88%

“…First, electron microscopy, used alone to track individual microtubules or coupled to chromosome micromanipulation, has shown that minus ends of nonkinetochore microtubules are physically linked to kinetochore microtubules near spindle poles (Nicklas et al, 1982;Mastronarde et al, 1993). Next, immunogold electron microscopy indicates that NuMA cross-links spindle microtubules at spindle poles (Dionne et al, 1999) and in vitro experiments show that NuMA can bind directly to microtubules and cross-link adjacent microtubules into bundles (Merdes et al, 1996;Haren and Merdes, 2002). Finally, it was recently shown that kinetochore fiber minus ends can be recruited to spindle poles through a mechanism that relies on the activity of NuMA to cross-link kinetochore microtubule minus ends to nonkinetochore astral microtubules (Khodjakov et al, 2003).…”

Section: Spindle Organizationmentioning

confidence: 99%

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A Functional Relationship between NuMA and Kid Is Involved in Both Spindle Organization and Chromosome Alignment in Vertebrate Cells

Levesque

Howard

Gordon

et al. 2003

MBoC

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We examined spindle morphology and chromosome alignment in vertebrate cells after simultaneous perturbation of the chromokinesin Kid and either NuMA, CENP-E, or HSET. Spindle morphology and chromosome alignment after simultaneous perturbation of Kid and either HSET or CENP-E were no different from when either HSET or CENP-E was perturbed alone. However, short bipolar spindles with organized poles formed after perturbation of both Kid and NuMA in stark contrast to splayed spindle poles observed after perturbation of NuMA alone. Spindles were disorganized if Kid, NuMA, and HSET were perturbed, indicating that HSET is sufficient for spindle organization in the absence of Kid and NuMA function. In addition, chromosomes failed to align efficiently at the spindle equator after simultaneous perturbation of Kid and NuMA despite appropriate kinetochore-microtubule interactions that generated chromosome movement at normal velocities. These data indicate that a functional relationship between the chromokinesin Kid and the spindle pole organizing protein NuMA influences spindle morphology, and we propose that this occurs because NuMA forms functional linkages between kinetochore and nonkinetochore microtubules at spindle poles. In addition, these data show that both Kid and NuMA contribute to chromosome alignment in mammalian cells.

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Section: Spindle Organizationmentioning

confidence: 72%

Section: Discussionmentioning

confidence: 88%

Section: Spindle Organizationmentioning

confidence: 99%

See 1 more Smart Citation

A Functional Relationship between NuMA and Kid Is Involved in Both Spindle Organization and Chromosome Alignment in Vertebrate Cells

Levesque

Howard

Gordon

et al. 2003

MBoC

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“…However, their ability to directly anchor components of the ␥ TuRC and thus serve as molecular scaffolds for tethering these complexes to centrosomes has not been demonstrated. These include the centrosome proteins Asp (do Carmo Avides and Glover, 1999), NuMA (Merdes et al, 1996), TPX-2 (Wittmann et al, 2000;Garrett et al, 2002), SPD-5 (Hamill et al, 2002), PCM-1 (Dammermann and Merdes, 2002), Sas-4 (Kirkham et al, 2003) centrosomin (Megraw et al, 1999;Terada et al, 2003), and several regulatory molecules, including Aurora A (Hannak et al, 2001;Giet et al, 2002), Polo (Lane and Nigg, 1996;Barbosa et al, 2000), PP1 (Katayama et al, 2001), and PP4 (Sumiyoshi et al, 2002).…”

Section: Other Proteins Involved In Centrosomal ␥ Turc Anchoring and mentioning

confidence: 99%

Mitosis-specific Anchoring of γ Tubulin Complexes by Pericentrin Controls Spindle Organization and Mitotic Entry

Zimmerman

Sillibourne

Rosa

et al. 2004

MBoC

275

258

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Microtubule nucleation is the best known function of centrosomes. Centrosomal microtubule nucleation is mediated primarily by ␥ tubulin ring complexes (␥ TuRCs). However, little is known about the molecules that anchor these complexes to centrosomes. In this study, we show that the centrosomal coiled-coil protein pericentrin anchors ␥ TuRCs at spindle poles through an interaction with ␥ tubulin complex proteins 2 and 3 (GCP2/3). Pericentrin silencing by small interfering RNAs in somatic cells disrupted ␥ tubulin localization and spindle organization in mitosis but had no effect on ␥ tubulin localization or microtubule organization in interphase cells. Similarly, overexpression of the GCP2/3 binding domain of pericentrin disrupted the endogenous pericentrin-␥ TuRC interaction and perturbed astral microtubules and spindle bipolarity. When added to Xenopus mitotic extracts, this domain uncoupled ␥ TuRCs from centrosomes, inhibited microtubule aster assembly, and induced rapid disassembly of preassembled asters. All phenotypes were significantly reduced in a pericentrin mutant with diminished GCP2/3 binding and were specific for mitotic centrosomal asters as we observed little effect on interphase asters or on asters assembled by the Ran-mediated centrosome-independent pathway. Additionally, pericentrin silencing or overexpression induced G2/antephase arrest followed by apoptosis in many but not all cell types. We conclude that pericentrin anchoring of ␥ tubulin complexes at centrosomes in mitotic cells is required for proper spindle organization and that loss of this anchoring mechanism elicits a checkpoint response that prevents mitotic entry and triggers apoptotic cell death. INTRODUCTIONThe centrosome is the primary microtubule-organizing center in animal cells. At the centrosome core is a pair of barrel-shaped microtubule assemblies, the centrioles (Doxsey, 2001). Centrioles are capable of self-assembly (Marshall et al., 2001;Khodjakov et al., 2002) and can serve as templates for recruitment and organization of the surrounding pericentriolar matrix (Bobinnec et al., 1998;Kirkham et al., 2003). The pericentriolar material or centrosome matrix contains a high proportion of coiled coil proteins and is the site of microtubule nucleation. Within the matrix are large protein complexes of ␥ tubulin and associated proteins that have a ring-like structure and mediate the nucleation of microtubules called ␥ tubulin ring complexes or ␥ TuRCs (Moritz et al., 1995a;Zheng et al., 1995). Other proteins may share the ability to nucleate microtubules because centrosomes can organize microtubules in the absence of functional ␥ tubulin (Sampaio et al., 2001;Strome et al., 2001;Hannak et al., 2002).During cell cycle progression, centrosomes "mature" by recruiting additional ␥ TuRCs and several other proteins, resulting in an increase in the nucleation capacity of the centrosome (reviewed in Blagden and Glover, 2003). However, we still know very little about proteins that directly anchor ␥ TuRCs to centrosomes in vertebrate cells. ...

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“…A number of scaffold proteins have been identified that associate with the mitotic spindle and play important roles in spindle assembly and cell cycle control. Examples of these proteins include γ-tubulin, NuMA, TPX2, dynactin and nucleolar spindle-associated protein (NuSAP) (Merdes et al, 1996;Hetzer et al, 2000). NuSAP is a wellknown microtubule-associated protein.…”

Section: Introductionmentioning

confidence: 99%

Nusap1 is essential for neural crest cell migration in zebrafish

Nie

Wang

et al. 2010

Protein Cell

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Microtubules play important roles in mitotic spindle assembly and chromosome segregation to maintain normal cell cycle progression. A number of microtubule-associated proteins have been identified in epithelial and neural cell cultures; however, their physiological significance is not well characterized due to the lack of appropriate in vivo animal models. Nucleolar spindleassociated protein (NuSAP) is a microtubule-binding protein and is reported to be involved in mitosis by cell culture studies. In this report, we identified the zebrafish homologue of human NuSAP and investigated its expression profile and functions. Using in situ hybridization, we demonstrated that transcripts of zebrafish nusap1 are specifically expressed in the retina, forebrain, hindbrain and neural crest. When the in vivo expression of nusap1 was knocked down through antisense oligonucleotide morpholino technology, the morphants of nusap1 showed impaired morphogenesis in the trunk and yolk extension, implying the involvement of Nusap1 in cell migration. Mechanistic studies revealed that nusap1 morphants have an altered expression pattern of neural crest markers crestin and sox9b, but normal expression of blood vessel and notochord markers gata1 and shh. In addition, nusap1 mRNA injection caused serious apoptosis in retina and hindbrain tissue, and these phenotypes can be rescued by co-injection of morpholino against nusap1. These observations not only suggest a role for Nusap1 in connecting apoptosis with cell migration, but also provide strong evidences that Nusap1 is potentially involved in morphogenesis in vertebrates.

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A Complex of NuMA and Cytoplasmic Dynein Is Essential for Mitotic Spindle Assembly

Cited by 554 publications

References 43 publications

A Functional Relationship between NuMA and Kid Is Involved in Both Spindle Organization and Chromosome Alignment in Vertebrate Cells

A Functional Relationship between NuMA and Kid Is Involved in Both Spindle Organization and Chromosome Alignment in Vertebrate Cells

Mitosis-specific Anchoring of γ Tubulin Complexes by Pericentrin Controls Spindle Organization and Mitotic Entry

Nusap1 is essential for neural crest cell migration in zebrafish

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