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Cited by 290Microtubule and mitosis dynamicsNuclear Structure and FunctionNeurogenesis and neuroplasticity mechanisms

Opposing motor activities are required for the organization of the mammalian mitotic spindle pole.

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Gaglio, T, Saredi, A, Bingham, J B, Hasbani, M J, Gill, S R, Schroer, T A, Compton, D A·1996·The Journal of cell biology
DOI 10.1083/jcb.135.2.399PMC PMC2121053
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Abstract

We use both in vitro and in vivo approaches to examine the roles of Eg5 (kinesin-related protein), cytoplasmic dynein, and dynactin in the organization of the microtubules and the localization of NuMA (Nu-clear protein that associates with the Mitotic Apparatus) at the polar ends of the mammalian mitotic spindle. Perturbation of the function of Eg5 through either immunodepletion from a cell free system for assembly of mitotic asters or antibody microinjection into cultured cells leads to organized astral microtubule arrays with expanded polar regions in which the minus ends of the microtubules emanate from a ring-like structure that contains NuMA. Conversely, perturbation of the function of cytoplasmic dynein or dynactin through either specific immunodepletition from the cell free system or expression of a dominant negative subunit of dynactin in cultured cells results in the complete lack of organization of microtubules and the failure to efficiently concentrate the NuMA protein despite its association with the microtubules. Simultaneous immunodepletion of these proteins from the cell free system for mitotic aster assembly indicates that the plus end-directed activity of Eg5 antagonizes the minus end-directed activity of cytoplasmic dynein and a minus end-directed activity associated with NuMA during the organization of the microtubules into a morphologic pole. Taken together, these results demonstrate that the unique organization of the minus ends of microtubules and the localization of NuMA at the polar ends of the mammalian mitotic spindle can be accomplished in a centrosome-independent manner by the opposing activities of plus end- and minus end-directed motors.

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  • The first role of minus end-directed kinesins is to generate counteracting forces against plus end-directed motors to maintain the spindle.

    “The first role is to generate counteracting forces against plus end-directed motors to maintain the spindle (1)(2)(3)(4).”

    From Diffusion and Directed Movement (2008) · cites this paper as [2]

    The paper reports that minus end-directed motor activity counteracts plus end-directed Eg5 during mitotic aster and spindle-pole organization (p. 399; Fig. 12, p. 410). It identifies cytoplasmic dynein as one minus end-directed motor and proposes, but does not establish, that another may be kinesin-related (p. 411). It therefore does not establish this as a role of minus end-directed kinesins generally or demonstrate spindle maintenance specifically. (LLM, conf=0.86)

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Authors
Gaglio, T., Saredi, A., Bingham, J. B., Hasbani, M. J., Gill, S. R., Schroer, T. A., & Compton, D. A.
Affiliations
Dartmouth College (US)
ORCID
Venue
The Journal of cell biology
Publisher
Rockefeller University Press
Published
1996-10-15
Volume
135
Issue
2
Pages
399-414
ISSN
  • 0021-9525 (ISSN-L)
  • 1540-8140
DOI
10.1083/jcb.135.2.399
PMC
PMC2121053
OpenAlex
W2006876135
Cited by
290
References
92
Citations by year
2025: 3 · 2024: 3 · 2023: 4 · 2022: 2 · 2021: 8 · 2020: 5 · 2019: 6 · 2018: 6 · 2017: 9 · 2016: 2 · 2015: 9 · 2014: 7 · 2013: 7 · 2012: 9
FWCI
9.19
Type
journal article
Language
en
Open access
Licence
cc-by-nc-sa
Locations
  • The Journal of Cell Biology (publishedVersion)
  • PubMed Central (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
Funders
National Institute of General Medical Sciences (R01GM044589)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nuclear Structure and Function
  • Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
Keywords
mitotic spindle poles, Eg5, kinesin-related protein, cytoplasmic dynein, dynactin, NUMA, microtubule organization, astral microtubules
MeSH terms
Dynactin Complex, Animals, Antibodies, Monoclonal, Cell Line, Cell-Free System, Chlorocebus aethiops, Chickens, Dyneins, HeLa Cells, Humans, Microtubule Proteins, Microtubule-Associated Proteins, Mitosis, Spindle Apparatus, Models, Biological, Nuclear Proteins, Nucleolus Organizer Region, Recombinant Proteins, Kinesins, Cell Cycle Proteins, Xenopus Proteins, Nuclear Matrix-Associated Proteins, Antigens, Nuclear

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