Inducible phosphorylation of cortactin is not necessary for cortactin-mediated actin polymerisation.
Cortactin is an SH3 domain-containing protein that contributes to the formation of dynamic cortical actin-associated structures, such as lamellipodia and membrane ruffles. It was originally identified as a substrate for the protein kinase Src; however, the role of tyrosine phosphorylation in the translocation of cortactin to the cell periphery and in the subsequent actin polymerisation is still unclear. Recently, two serine/threonine kinases, Pak1 and Erk, have been implicated in the regulation of cortactin. Therefore, we systematically investigated whether phosphorylation on either tyrosine or serine/threonine residues is necessary for cortactin function. In COS7 cells over-expressing Vav2 or treated with EGF, we could not detect tyrosine phosphorylation, although cortactin was translocated to cell periphery and induced membrane ruffle formation. In addition, the selective MEK inhibitor, PD98059, did not influence in vivo the ability of cortactin to bind to and induce membrane ruffles upon Vav2 over-expression or short-term EGF treatment. Finally, using a constitutively active Pak1 mutant, Pak1 T423E, we showed that Pak1 is not capable of phosphorylating cortactin either in vitro or in COS7 cells. These results suggest that cortactin-mediated actin polymerisation at cell periphery requires only Rac activation but neither tyrosine nor serine/threonine phosphorylation.
Illés
András
A
Department of Medical Chemistry, Semmelweis University Medical School, 9 Puskin Street, 1088 Budapest, Hungary.
Enyedi
Balázs
B
Tamás
Péter
P
Balázs
Annamária
A
Bogel
Gábor
G
Buday
László
L
D000199:Actins / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D051356:Cortactin / Q000378:metabolism / Q000502:physiology*; D004815:Epidermal Growth Factor / Q000378:metabolism; D006801:Humans; D020935:MAP Kinase Signaling System; D010766:Phosphorylation; D011108:Polymers / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D051567:Proto-Oncogene Proteins c-vav / Q000378:metabolism; D054462:p21-Activated Kinases; D020744:rac GTP-Binding Proteins / Q000378:metabolism*
D000199:Actins; D051356:Cortactin; D011108:Polymers; D051567:Proto-Oncogene Proteins c-vav; C093651:VAV2 protein, human; D004815:Epidermal Growth Factor; C515903:PAK1 protein, human; D017346:Protein Serine-Threonine Kinases; D054462:p21-Activated Kinases; D020744:rac GTP-Binding Proteins