Felic (CIP4b), a novel binding partner with the Src kinase Lyn and Cdc42, localizes to the phagocytic cup.
Through its Src homology 3 (SH3) and SH2 domains, the Src kinase Lyn interacts with a small number of phosphoproteins, such as Shc, Cbl, and Vav, which regulate cell cycle and the cytoskeleton. Using Lyn's Unique, SH3, and SH2 domains as bait in a yeast 2-hybrid screen, we isolated a novel gene product with features of a scaffolding protein. We named it Felic because it contains a domain homologous to the tyrosine kinase Fes and the cytoskeletal protein ezrin and forms a Lyn interaction with the GTPase Cdc42 (Felic). Felic was expressed in both hematopoietic and nonhematopoietic tissues. Because it represents an alternative splice product related to the Cdc42-interacting protein 4, CIP4, we also refer to Felic as CIP4b. Felic contains an SH3 recognition site RXPXXP and multiple tyrosine residues. In insulin or serum-stimulated HEK293 cells, Felic became tyrosine phosphorylated. Like CIP4, Felic associated with Cdc42 in its activated form only. Unlike CIP4, Felic does not possess a C-terminal SH3 domain. Coprecipitation studies show that Felic bound to Lyn or activated forms of Cdc42. Overexpression of Felic or CIP4 inhibited NIH 3T3 cell invasiveness in a Matrigel assay. Because Lyn and Cdc42 are involved in phagocytosis, we examined the distribution of Felic in RAW macrophages during particle ingestion. Felic was recruited more efficiently than CIP4 to the phagocytic cups. Altogether, these data suggest that CIP4/Felic constitute a novel family of cytoskeletal scaffolding proteins, integrating Src and Cdc42 pathways. The absence of an SH3 domain in Felic provides a structural basis for functional differences.
Dombrosky-Ferlan
Patrice
P
Department of Pediatrics, University of Pittsburgh, PA, USA.
Grishin
Anatoly
A
Botelho
Roberto J
RJ
Sampson
Matthew
M
Wang
Lin
L
Rudert
William A
WA
Grinstein
Sergio
S
Corey
Seth J
SJ
D000595:Amino Acid Sequence; D002465:Cell Movement / Q000187:drug effects; D006801:Humans; D008264:Macrophages / Q000378:metabolism / Q000648:ultrastructure; D008869:Microtubule-Associated Proteins / Q000378:metabolism* / Q000502:physiology; D015778:Minor Histocompatibility Antigens; D008969:Molecular Sequence Data; D010588:Phagosomes / Q000378:metabolism; D011485:Protein Binding; D020033:Protein Isoforms / Q000378:metabolism / Q000502:physiology; D014407:Tumor Cells, Cultured; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism* / Q000502:physiology; D018909:src Homology Domains; D019061:src-Family Kinases / Q000378:metabolism*
D008869:Microtubule-Associated Proteins; D015778:Minor Histocompatibility Antigens; D020033:Protein Isoforms; C406285:TRIP10 protein, human; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases; D020764:cdc42 GTP-Binding Protein