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PMID:16968699

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Citation

Han, JW, Leeper, L, Rivero, F and Chung, CY (2006) Role of RacC for the regulation of WASP and phosphatidylinositol 3-kinase during chemotaxis of Dictyostelium. J. Biol. Chem. 281:35224-34

Abstract

WASP family proteins are key players for connecting multiple signaling pathways to F-actin polymerization. To dissect the highly integrated signaling pathways controlling WASP activity, we identified a Rac protein that binds to the GTPase binding domain of WASP. Using two-hybrid and FRET-based functional assays, we identified RacC as a major regulator of WASP. RacC stimulates F-actin assembly in cell-free systems in a WASP-dependent manner. A FRET-based microscopy approach showed local activation of RacC at the leading edge of chemotaxing cells. Cells overexpressing RacC exhibit a significant increase in the level of F-actin polymerization upon cAMP stimulation, which can be blocked by a phosphatidylinositol (PI) 3-kinase inhibitor. Membrane translocation of PI 3-kinase and PI 3,4,5-trisphosphate reporter is absent in racC null cells. Cells overexpressing dominant negative RacC mutants and racC null cells move at a significantly slower speed and show a poor directionality during chemotaxis. Our results suggest that RacC plays an important role in PI 3-kinase activation and WASP activation for dynamic regulation of F-actin assembly during Dictyostelium chemotaxis.

Links

PubMed PMC2853593 Online version:10.1074/jbc.M605997200

Keywords

Animals; Chemotaxis/physiology; Dictyostelium/cytology; Dictyostelium/metabolism; Humans; Phosphatidylinositol 3-Kinases/metabolism; Phosphatidylinositol Phosphates/metabolism; Wiskott-Aldrich Syndrome Protein/metabolism; rac GTP-Binding Proteins/metabolism

Significance

Annotations

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

DICDI:RAC1A

involved_in

GO:0030838: positive regulation of actin filament polymerization

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:RAC1B

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9GSG9

F

Seeded From UniProt

complete

DICDI:RAC1B

involved_in

GO:0030838: positive regulation of actin filament polymerization

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:RACA

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9GSG9

F

Seeded From UniProt

complete

DICDI:RACB

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9GSG9

F

Seeded From UniProt

complete

DICDI:RACC

acts_upstream_of_or_within

GO:0000281: mitotic cytokinesis

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:RACC

acts_upstream_of_or_within

GO:0016477: cell migration

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:RACC

enables

GO:0019904: protein domain specific binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

DICDI:RACC

part_of

GO:0031410: cytoplasmic vesicle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DICDI:RACC

involved_in

GO:0030838: positive regulation of actin filament polymerization

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:RACC

acts_upstream_of_or_within

GO:0030838: positive regulation of actin filament polymerization

ECO:0000316: genetic interaction evidence used in manual assertion

UniProtKB:Q9GSG9

P

Seeded From UniProt

complete

DICDI:RACC

acts_upstream_of_or_within

GO:0014065: phosphatidylinositol 3-kinase signaling

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:RACC

acts_upstream_of_or_within

GO:0006935: chemotaxis

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:RACC

is_active_in

GO:0031252: cell leading edge

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DICDI:RACC

acts_upstream_of_or_within

GO:0045184: establishment of protein localization

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:KRAC

located_in

GO:0005938: cell cortex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DICDI:PI3K2

located_in

GO:0005938: cell cortex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:CDC42

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9GSG9

F

Seeded From UniProt

complete

HUMAN:RAC1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9GSG9

F

Seeded From UniProt

complete

DICDI:Q9GSG9

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P34148

F

Seeded From UniProt

complete

DICDI:Q9GSG9

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P34147

F

Seeded From UniProt

complete

DICDI:Q9GSG9

enables

GO:0031267: small GTPase binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P34145

F

Seeded From UniProt

complete

DICDI:Q9GSG9

enables

GO:0031267: small GTPase binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P63000

F

Seeded From UniProt

complete

DICDI:Q9GSG9

enables

GO:0031267: small GTPase binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P60953

F

Seeded From UniProt

complete

DICDI:KRAC

part_of

GO:0005938: cell cortex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

See also

References

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