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PMID:15548599

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Citation

Hotulainen, P, Paunola, E, Vartiainen, MK and Lappalainen, P (2005) Actin-depolymerizing factor and cofilin-1 play overlapping roles in promoting rapid F-actin depolymerization in mammalian nonmuscle cells. Mol. Biol. Cell 16:649-64

Abstract

Actin-depolymerizing factor (ADF)/cofilins are small actin-binding proteins found in all eukaryotes. In vitro, ADF/cofilins promote actin dynamics by depolymerizing and severing actin filaments. However, whether ADF/cofilins contribute to actin dynamics in cells by disassembling "old" actin filaments or by promoting actin filament assembly through their severing activity is a matter of controversy. Analysis of mammalian ADF/cofilins is further complicated by the presence of multiple isoforms, which may contribute to actin dynamics by different mechanisms. We show that two isoforms, ADF and cofilin-1, are expressed in mouse NIH 3T3, B16F1, and Neuro 2A cells. Depleting cofilin-1 and/or ADF by siRNA leads to an accumulation of F-actin and to an increase in cell size. Cofilin-1 and ADF seem to play overlapping roles in cells, because the knockdown phenotype of either protein could be rescued by overexpression of the other one. Cofilin-1 and ADF knockdown cells also had defects in cell motility and cytokinesis, and these defects were most pronounced when both ADF and cofilin-1 were depleted. Fluorescence recovery after photobleaching analysis and studies with an actin monomer-sequestering drug, latrunculin-A, demonstrated that these phenotypes arose from diminished actin filament depolymerization rates. These data suggest that mammalian ADF and cofilin-1 promote cytoskeletal dynamics by depolymerizing actin filaments and that this activity is critical for several processes such as cytokinesis and cell motility.

Links

PubMed PMC545901 Online version:10.1091/mbc.E04-07-0555

Keywords

Actins/drug effects; Actins/genetics; Actins/metabolism; Animals; Bicyclo Compounds, Heterocyclic/pharmacology; Blotting, Western; Cell Line, Tumor; Cell Movement/genetics; Cell Size; Cofilin 1; Cytokinesis/genetics; Destrin; Fibroblasts/metabolism; Fibronectins/metabolism; Fluorescein-5-isothiocyanate; Fluorescence Recovery After Photobleaching; Fluorescent Antibody Technique; Fluorescent Dyes; Gene Silencing; Indoles; Kinetics; Melanoma, Experimental/metabolism; Mice; Microfilament Proteins/genetics; Microfilament Proteins/metabolism; Microscopy, Video; NIH 3T3 Cells; Protein Isoforms/genetics; Protein Isoforms/metabolism; RNA, Small Interfering/metabolism; Stress Fibers/metabolism; Thiazoles/pharmacology; Thiazolidines

Significance

Annotations

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

MOUSE:G3UW82

located_in

GO:0005826: actomyosin contractile ring

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:I6L963

located_in

GO:0005826: actomyosin contractile ring

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:COF1

located_in

GO:0005737: cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:COF1

located_in

GO:0030864: cortical actin cytoskeleton

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:Q1WNQ9

located_in

GO:0005826: actomyosin contractile ring

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:DEST

located_in

GO:0005737: cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:DEST

located_in

GO:0030864: cortical actin cytoskeleton

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:DEST

acts_upstream_of_or_within

GO:0030836: positive regulation of actin filament depolymerization

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:Q9WUS5

located_in

GO:0005826: actomyosin contractile ring

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:COF1

acts_upstream_of_or_within

GO:0000281: mitotic cytokinesis

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:COF1

part_of

GO:0005737: cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

MOUSE:COF1

acts_upstream_of_or_within

GO:0030836: positive regulation of actin filament depolymerization

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

MOUSE:COF1

part_of

GO:0030864: cortical actin cytoskeleton

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

See also

References

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