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PMID: 8241568 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Gradients in the concentration and assembly of myosin II in living fibroblasts during locomotion and fiber transport.

Molecular biology of the cell ·Vol. 4 ·No. 8 ·1993-08-00 ·Pages 819-36

Kolega J, Taylor DL

Abstract

Assembly and motor activity of myosin II affect shape, contractility, and locomotion of nonmuscle cells. We used fluorescent analogues and imaging techniques to elucidate the state of assembly and three-dimensional distribution of myosin II in living Swiss 3T3 fibroblasts. An analogue of myosin II that was covalently cross-linked in the 10S conformation and unable to assemble served as an indicator of the cytoplasmic volume accessible to 10S myosin II. Ratio-imaging of an analogue that can undergo 10S-->6S conversion versus the volume indicator revealed localized concentration of assembly-competent myosin II. In stationary serum-deprived cells and in cells locomoting at the edge of a wound, it was most concentrated in the peripheral cytoplasm, where fibers containing myosin II assemble, and least concentrated in the perinuclear cytoplasm, where they disassemble. Furthermore, fluorescence photobleaching recovery showed myosin II to be less mobile in the periphery than in perinuclear cytoplasm. These results indicate a gradient in the assembly of myosin II. Three-dimensional microscopy of living cells revealed that fibers containing myosin II were localized in the cortical cytoplasm, whereas myosin II was diffusely distributed in the deeper cytoplasm, suggesting that myosin II is assembled preferentially near the cell surface. Localized protein phosphorylation may play a role, because a kinase inhibitor, staurosporine, abolished the gradient of myosin II assembly.

MeSH Terms
3T3 Cells Alkaloids/pharmacology Animals Biological Transport Cell Movement Chickens Fibroblasts/cytology,metabolism,ultrastructure Mice Microscopy, Fluorescence Myosins/chemistry,metabolism Protein Conformation Staurosporine
Chemicals
Alkaloids Myosins Staurosporine
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Kolega J
Center for Light Microscope Imaging and Biotechnology, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213.
Taylor D L
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.4.8.819
Published
1993-08-00
Pages
819-36
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC300995
Subset
IM
Grants
NIAMS NIH HHS · AR-32461 · United States
External Links
PubMed source DOI: 10.1091/mbc.4.8.819 PMC Full Text View journal details
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