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PMID:9852149

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Citation

Buss, F, Kendrick-Jones, J, Lionne, C, Knight, AE, Côté, GP and Paul Luzio, J (1998) The localization of myosin VI at the golgi complex and leading edge of fibroblasts and its phosphorylation and recruitment into membrane ruffles of A431 cells after growth factor stimulation. J. Cell Biol. 143:1535-45

Abstract

Myosin VI is an unconventional myosin that may play a role in vesicular membrane traffic through actin rich regions of the cytoplasm in eukaryotic cells. In this study we have cloned and sequenced a cDNA encoding a chicken intestinal brush border myosin VI. Polyclonal antisera were raised to bacterially expressed fragments of this myosin VI. The affinity purified antibodies were highly specific for myosin VI by immunoblotting and immunoprecipitation and were used to study the localization of the protein by immunofluorescence and immunoelectron microscopy. It was found that in NRK and A431 cells, myosin VI was associated with both the Golgi complex and the leading, ruffling edge of the cell as well as being present in a cytosolic pool. In A431 cells in which cell surface ruffling was stimulated by EGF, myosin VI was phosphorylated and recruited into the newly formed ruffles along with ezrin and myosin V. In vitro experiments suggested that a p21-activated kinase (PAK) might be the kinase responsible for phosphorylation in the motor domain. These results strongly support a role for myosin VI in membrane traffic on secretory and endocytic pathways.

Links

PubMed PMC2132970

Keywords

Actins/metabolism; Amino Acid Sequence; Animals; Cell Line; Cell Membrane/drug effects; Cell Membrane/metabolism; Cell Membrane/ultrastructure; Chickens; Endocytosis; Epidermal Growth Factor/pharmacology; Fibroblasts/metabolism; Fibroblasts/ultrastructure; Golgi Apparatus/drug effects; Golgi Apparatus/metabolism; Golgi Apparatus/ultrastructure; Humans; Intracellular Membranes/drug effects; Intracellular Membranes/metabolism; Intracellular Membranes/ultrastructure; Kidney/metabolism; Kidney/ultrastructure; Liver/metabolism; Liver/ultrastructure; Microscopy, Immunoelectron; Microvilli/metabolism; Microvilli/ultrastructure; Molecular Sequence Data; Myosins/chemistry; Myosins/genetics; Myosins/metabolism; Phosphorylation; Protein-Serine-Threonine Kinases/metabolism; Rats; Sequence Alignment; Sequence Homology, Amino Acid; Tumor Cells, Cultured; p21-Activated Kinases

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

RAT:D4A5I9

located_in

GO:0032587: ruffle membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:D4A5I9

located_in

GO:0005794: Golgi apparatus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:D4A5I9

located_in

GO:0043231: intracellular membrane-bounded organelle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:EZRI

located_in

GO:0001726: ruffle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:EZRI

located_in

GO:0005829: cytosol

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MYO6

located_in

GO:0001726: ruffle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MYO6

located_in

GO:0048471: perinuclear region of cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MYO6

located_in

GO:0005737: cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MYO5A

located_in

GO:0001726: ruffle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MYO5A

located_in

GO:0005737: cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:PAK1

involved_in

GO:0006468: protein phosphorylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

HUMAN:PAK1

enables

GO:0004672: protein kinase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:EZRI

part_of

GO:0005829: cytosol

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:EZRI

part_of

GO:0001726: ruffle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MYO6

part_of

GO:0001726: ruffle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MYO6

part_of

GO:0005737: cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MYO6

part_of

GO:0031941: filamentous actin

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MYO6

part_of

GO:0048471: perinuclear region of cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:MYO6

enables

GO:0051015: actin filament binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete


See also

References

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