Cortactin is required for integrin-mediated cell spreading.
Cortactin is an SH3 domain-containing protein that contributes to the formation of dynamic cortical actin-associated structures, such as lamellipodia and membrane ruffles. Here we show that expression of either the GFP-tagged N-terminal or the C-teminal halves of cortactin inhibits significantly the spreading of COS7 cells on fibronectin. Introducing inactivating point mutation into the SH3 domain of the C-terminal half of cortactin suspends the dominant negative effect of the construct. In addition, a vector-based RNA interference was used to knock-down endogenous level of cortactin in cells. We demonstrate that cortactin deficient cells were not able to spread. These results suggest that cortactin is required for integrin-mediated signalling pathways.
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
Melinda
Lukács
Buday
László
L
D000818:Animals; D019556:COS Cells; D002448:Cell Adhesion; D002465:Cell Movement / Q000235:genetics; D002522:Chlorocebus aethiops; D051356:Cortactin / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D005353:Fibronectins / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D016023:Integrins / Q000378:metabolism*; D017354:Point Mutation; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics; D015533:Transcriptional Activation
D051356:Cortactin; D005353:Fibronectins; D016023:Integrins; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins