Myosin Dynamics in Live Dictyostelium Cells

Conventional myosin plays a key role in the cytoskeletal reorganization necessary for cytokinesis, migration, and morphological changes associated with development in nonmuscle cells. We have made a fusion between the green fluorescent protein (GFP) and the Dictyostelium discoideum myosin heavy chai...

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Published inProceedings of the National Academy of Sciences - PNAS Vol. 93; no. 1; pp. 443 - 446
Main Authors Moores, Sheri L., Sabry, James H., Spudich, James A.
Format Journal Article
LanguageEnglish
Published United States National Academy of Sciences of the United States of America 09.01.1996
National Acad Sciences
National Academy of Sciences
Subjects
Adenosine triphosphatases
Animals
Base Sequence
Cell Division
Cell lines
Cell Movement
Cells
Cellular biology
Cellular immunity
Cytokinesis
Dictyostelium - physiology
Dictyostelium discoideum
DNA Primers - chemistry
Fluorescence
Gels
Green Fluorescent Proteins
Imaging
Luminescent Proteins
Lymphocytes
Microscopy, Fluorescence
Molecular Sequence Data
Muscular system
Myosins - physiology
Plasmids
Proteins
Recombinant Fusion Proteins
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Summary:Conventional myosin plays a key role in the cytoskeletal reorganization necessary for cytokinesis, migration, and morphological changes associated with development in nonmuscle cells. We have made a fusion between the green fluorescent protein (GFP) and the Dictyostelium discoideum myosin heavy chain (GFP-myosin). The unique Dictyostelium system allows us to test the GFP-tagged myosin for activity both in vivo and in vitro. Expression of GFP-myosin rescues all myosin null cell defects. Additionally, GFP-myosin purified from these cells exhibits the same ATPase activities and in vitro motility as wild-type myosin. GFP-myosin is concentrated in the cleavage furrow during cytokinesis and in the posterior cortex of migrating cells. Surprisingly, GFP-myosin concentration increases transiently in the tips of retracting pseudopods. Contrary to previous thinking, this suggests that conventional myosin may play an important role in the dynamics of pseudopods as well as filopodia, lamellipodia, and other cellular protrusions.
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ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.93.1.443