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PMID: 11756470 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Cytoplasmic dynein/dynactin drives kinetochore protein transport to the spindle poles and has a role in mitotic spindle checkpoint inactivation.

The Journal of cell biology ·Vol. 155 ·No. 7 ·2001-12-24 ·Pages 1159-72

Howell BJ, McEwen BF, Canman JC, Hoffman DB, Farrar EM, Rieder CL, Salmon ED

Abstract

We discovered that many proteins located in the kinetochore outer domain, but not the inner core, are depleted from kinetochores and accumulate at spindle poles when ATP production is suppressed in PtK1 cells, and that microtubule depolymerization inhibits this process. These proteins include the microtubule motors CENP-E and cytoplasmic dynein, and proteins involved with the mitotic spindle checkpoint, Mad2, Bub1R, and the 3F3/2 phosphoantigen. Depletion of these components did not disrupt kinetochore outer domain structure or alter metaphase kinetochore microtubule number. Inhibition of dynein/dynactin activity by microinjection in prometaphase with purified p50 "dynamitin" protein or concentrated 70.1 anti-dynein antibody blocked outer domain protein transport to the spindle poles, prevented Mad2 depletion from kinetochores despite normal kinetochore microtubule numbers, reduced metaphase kinetochore tension by 40%, and induced a mitotic block at metaphase. Dynein/dynactin inhibition did not block chromosome congression to the spindle equator in prometaphase, or segregation to the poles in anaphase when the spindle checkpoint was inactivated by microinjection with Mad2 antibodies. Thus, a major function of dynein/dynactin in mitosis is in a kinetochore disassembly pathway that contributes to inactivation of the spindle checkpoint.

MeSH Terms
Animals Cell Line Cell Polarity Chromosomes Dyneins/metabolism Kinetochores/physiology Metaphase Spindle Apparatus/physiology
Chemicals
Dyneins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Howell B J
Department of Biology, University of North Carolina, Chapel Hill, NC 27599, USA. Bhowell@email.unc.edu
McEwen B F
Canman J C
Hoffman D B
Farrar E M
Rieder C L
Salmon E D
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200105093
Email
bhowell@email.unc.edu
Published
2001-12-24
Epub
2001-00-24
Pages
1159-72
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2199338
Subset
IM
Grants
NIGMS NIH HHS · GM-24364 · United States
NIGMS NIH HHS · R37 GM040198-18 · United States
NIGMS NIH HHS · GMS-R01-40198 · United States
NIGMS NIH HHS · R37 GM040198 · United States
NIGMS NIH HHS · R37 GM024364 · United States
NIGMS NIH HHS · R01 GM024364 · United States
External Links
PubMed source DOI: 10.1083/jcb.200105093 PMC Full Text View journal details
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