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PMID: 10811826 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Formation of spindle poles by dynein/dynactin-dependent transport of NuMA.

The Journal of cell biology ·Vol. 149 ·No. 4 ·2000-05-15 ·Pages 851-62

Merdes A, Heald R, Samejima K, Earnshaw WC, Cleveland DW

Abstract

NuMA is a large nuclear protein whose relocation to the spindle poles is required for bipolar mitotic spindle assembly. We show here that this process depends on directed NuMA transport toward microtubule minus ends powered by cytoplasmic dynein and its activator dynactin. Upon nuclear envelope breakdown, large cytoplasmic aggregates of green fluorescent protein (GFP)-tagged NuMA stream poleward along spindle fibers in association with the actin-related protein 1 (Arp1) protein of the dynactin complex and cytoplasmic dynein. Immunoprecipitations and gel filtration demonstrate the assembly of a reversible, mitosis-specific complex of NuMA with dynein and dynactin. NuMA transport is required for spindle pole assembly and maintenance, since disruption of the dynactin complex (by increasing the amount of the dynamitin subunit) or dynein function (with an antibody) strongly inhibits NuMA translocation and accumulation and disrupts spindle pole assembly.

MeSH Terms
Biological Transport Cell Compartmentation Dynactin Complex Dyneins/isolation & purification,metabolism Fluorescent Antibody Technique Green Fluorescent Proteins Luminescent Proteins/genetics,isolation & purification,metabolism Metaphase Microtubule-Associated Proteins/isolation & purification,metabolism Microtubules/metabolism Models, Biological Nuclear Envelope/metabolism Nuclear Proteins/genetics,isolation & purification,metabolism Recombinant Fusion Proteins/isolation & purification,metabolism Spindle Apparatus/metabolism
Chemicals
Dynactin Complex Luminescent Proteins Microtubule-Associated Proteins Nuclear Proteins Recombinant Fusion Proteins Green Fluorescent Proteins Dyneins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Merdes A
ICMB, University of Edinburgh, Scotland. [email protected]
Heald R
Samejima K
Earnshaw W C
Cleveland D W
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.149.4.851
Email
[email protected]
Published
2000-05-15
Pages
851-62
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2174573
Subset
IM
Grants
Wellcome Trust · 073915 · United Kingdom
NIGMS NIH HHS · GM 29513 · United States
External Links
PubMed source DOI: 10.1083/jcb.149.4.851 PMC Full Text View journal details
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