Opposite effects of overexpressed myosin Va or heavy meromyosin Va on vesicle distribution, cytoskeleton organization, and cell motility in nonmuscle cells

Robbin D Eppinga; I-Feng Peng; Jenny Li-Chun Lin; Chun-Fang Wu; Jim Jung-Ching Lin

doi:10.1002/cm.20255

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Journal article

Opposite effects of overexpressed myosin Va or heavy meromyosin Va on vesicle distribution, cytoskeleton organization, and cell motility in nonmuscle cells

Robbin D Eppinga, I-Feng Peng, Jenny Li-Chun Lin, Chun-Fang Wu and Jim Jung-Ching Lin

Cell motility and the cytoskeleton, Vol.65(3), pp.197-215

03/2008

DOI: 10.1002/cm.20255

PMID: 18044718

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Abstract

Myosin Va, an actin-based motor protein that transports intracellular cargos, can bundle actin in vitro. Whether myosin Va regulates cellular actin dynamics or cell migration remains unclear. To address this, we compared Chinese Hamster Ovary (CHO) cells that stably express GFP fused to either full length mouse myosin Va (GFP-M5) or heavy meromyosin Va (GFP-M5Delta). GFP-M5 and GFP-M5Delta co-immunoprecipitate with CHO myosin Va and serve as overexpression of wild-type and dominant negative mutants of myosin Va. Compared to non-expressing control cells, GFP-M5-overexpressing cells have peripheral endocytic vesicles, spread slowly after plating, as well as produce robust interior actin stress fibers, myosin II bundles, and focal adhesions. However, these cells display normal cell migration and lamellipodial dynamics. In contrast, GFP-M5Delta-expressing cells have perinuclear endocytic vesicles, produce thin interior actin and myosin bundles and contain no interior focal adhesions. In addition, these cells spread rapidly, migrate slowly and display reduced lamellipodial dynamics. Similarly, neurite outgrowth is compromised in neurons cultured from transgenic Drosophila that express M5Delta-dsRed and in neurons cultured from Drosophila that produce a tailless version of endogenous myosin V. Together, these data suggest that myosin Va overexpression induces actin bundles in vivo whereas the tailless version fails to bundle actin and disrupts cell motility.

Cell Proliferation

Immunoprecipitation

Cricetulus

Actins - metabolism

Neurons - cytology

Myosin Heavy Chains - metabolism

Recombinant Fusion Proteins - metabolism

Endocytosis

Myosin Type II - metabolism

Cell Division

Cell Shape

Neurons - metabolism

CHO Cells

Green Fluorescent Proteins - metabolism

Cricetinae

Myosin Type V - metabolism

Myosin Subfragments - metabolism

Mutant Proteins - metabolism

Protein Transport

Animals

Transport Vesicles - metabolism

Cytoskeleton - metabolism

Mice

Drosophila melanogaster

Cell Movement

Details

Title: Subtitle: Opposite effects of overexpressed myosin Va or heavy meromyosin Va on vesicle distribution, cytoskeleton organization, and cell motility in nonmuscle cells
Creators: Robbin D Eppinga - Department of Biological Sciences, University of Iowa, Iowa City, Iowa 52242-1324, USA
I-Feng Peng
Jenny Li-Chun Lin
Chun-Fang Wu
Jim Jung-Ching Lin
Resource Type: Journal article
Publication Details: Cell motility and the cytoskeleton, Vol.65(3), pp.197-215
Publisher: United States
DOI: 10.1002/cm.20255
PMID: 18044718
ISSN: 0886-1544
eISSN: 1097-0169
Grant note: HD18577 / NICHD NIH HHS
Language: English
Date published: 03/2008
Academic Unit: Iowa Neuroscience Institute; Biology
Record Identifier: 9984070414602771

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