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PMID: 1835460 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Two activators of microtubule-based vesicle transport.

The Journal of cell biology ·Vol. 115 ·No. 5 ·1991-12-00 ·Pages 1309-18

Schroer TA, Sheetz MP

Abstract

Cytoplasmic dynein purified by nucleotide dependent microtubule affinity has significant minus end-directed vesicle motor activity that decreases with each further purification step. Highly purified dynein causes membrane vesicles to bind but not move on microtubules. We exploited these observations to develop an assay for factors that, in combination with dynein, would permit minus end-directed vesicle motility. At each step of the purification, non-dynein fractions were recombined with dynein and assayed for vesicle motility. Two activating fractions were identified by this method. One, called Activator I, copurified with 20S dynein by velocity sedimentation but could be separated from it by ion exchange chromatography. Activator I increased only the frequency of dynein-driven vesicle movements. Activator II, sedimenting at 9S, increased both the frequency and velocity of vesicle transport and also supported plus end movements. Our results suggest that dynein-based motility is controlled at multiple levels and provide a preliminary characterization of two regulatory factors.

MeSH Terms
Animals Biological Transport Cell Membrane/metabolism Cells, Cultured Chick Embryo Dyneins/isolation & purification Electrophoresis, Polyacrylamide Gel Microtubules/metabolism
Chemicals
Dyneins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Schroer T A
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110.
Sheetz M P
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.115.5.1309
Published
1991-12-00
Pages
1309-18
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2289226
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.115.5.1309 PMC Full Text View journal details
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