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PMID:11950598

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Citation

Paris, S, Za, L, Sporchia, B and de Curtis, I (2002) Analysis of the subcellular distribution of avian p95-APP2, an ARF-GAP orthologous to mammalian paxillin kinase linker. Int. J. Biochem. Cell Biol. 34:826-37

Abstract

We describe here the identification and characterization of avian p95-APP2, a multi-domain protein of a recently identified family of ADP-ribosylation factor (ARF)-GTPase-activating proteins (GAPs) including mammalian G protein-coupled receptor kinases (GRK)-interactor 1 (GIT1), paxillin kinase linker (PKL), and GIT2, as well as avian p95-APP1. The p95-APP2 is eluted from Rac-GTP-gamma-S, but not from Rac-GDP-beta-S columns. As other members of the family, p95-APP2 has binding regions for the focal adhesion protein paxillin, and for the Rac exchanging factor PIX. Sequence comparison indicates that p95-APP2 is the avian orthologue of mammalian PKL. Expression studies showed a largely diffuse distribution of the full length p95-APP2, without evident effects on cell morphology. We observed a dramatic difference between the localization of the amino-terminal portion of the protein, including the ARF-GAP domain and the three ankyrin repeats, and the carboxy-terminal portion including the paxillin-binding site. Moreover, the expression of truncated carboxy-terminal polypeptides including both the PIX- and paxillin-binding regions leads to a marked localization of the protein together with paxillin at large vesicles. Comparison of the expression of corresponding ARF-GAP-deficient constructs from p95-APP2 and p95-APP1 shows their distribution at distinct endocytic compartments. Altogether, these data support a role of distinct members of this family of ARF-GAPs in the regulation of different steps of membrane traffic during cell motility, and suggest that p95-APP2 may shuttle between an intracellular compartment and the cell periphery, although, further work will be needed to address this point.

Links

PubMed

Keywords

ADP-Ribosylation Factors/chemistry; ADP-Ribosylation Factors/genetics; ADP-Ribosylation Factors/metabolism; Adaptor Proteins, Signal Transducing; Amino Acid Sequence; Animals; Base Sequence; Carrier Proteins/chemistry; Carrier Proteins/genetics; Carrier Proteins/metabolism; Cell Cycle Proteins; Cell Movement/physiology; Cells, Cultured; Chick Embryo; Chickens; Cloning, Molecular; DNA, Complementary/genetics; Fibroblasts/metabolism; GTPase-Activating Proteins/chemistry; GTPase-Activating Proteins/genetics; GTPase-Activating Proteins/metabolism; Humans; Macromolecular Substances; Molecular Sequence Data; Phosphoproteins; Protein Structure, Tertiary; Sequence Deletion; Species Specificity; Subcellular Fractions/metabolism

Significance

Annotations

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

RAT:ARHG7

contributes_to

GO:0031267: small GTPase binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q13153,UniProtKB:Q9DE64

F

  • has_input:(UniProtKB:O93466)

Seeded From UniProt

complete

RAT:ARHG7

part_of

GO:0032991: protein-containing complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:PAK1

contributes_to

GO:0031267: small GTPase binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O55043,UniProtKB:Q9DE64

F

  • has_input:(UniProtKB:O93466)

Seeded From UniProt

complete

CHICK:Q9DE64

contributes_to

GO:0031267: small GTPase binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O55043,UniProtKB:Q13153

F

  • has_input:(UniProtKB:O93466)

Seeded From UniProt

complete

CHICK:O93466

enables

GO:0044877: protein-containing complex binding

ECO:0000314: direct assay evidence used in manual assertion

F

  • has_input:(UniProtKB:O55043)
  • has_input:(UniProtKB:Q13153)
  • has_input:(UniProtKB:Q9DE64)

Seeded From UniProt

complete

CHICK:Q9DE64

contributes_to

GO:0048365: Rac GTPase binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O55043,UniProtKB:Q13153

F

has_input:(UniProtKB:O93466)

Seeded From UniProt

complete

CHICK:Q9DE64

part_of

GO:0032991: protein-containing complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:PAK1

part_of

GO:0032991: protein-containing complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

HUMAN:PAK1

contributes_to

GO:0048365: Rac GTPase binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O55043,UniProtKB:Q9DE64

F

has_input:(UniProtKB:O93466)

Seeded From UniProt

complete


See also

References

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