ASSOCIATION AND DIRECT ACTIVATION OF SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION1-ALPHA BY PLATELET-DERIVED GROWTH-FACTOR RECEPTOR

PDGF stimulates tyrosine phosphorylation of Janus kinase 1 (JAK1) and the signal transducer and activator of transcription 1 (STAT1 alpha), However, it is not known whether JAKs are required for STAT1 alpha pho sphorylation or if the PDGF receptor itself can directly tyrosine phos phorylate and activate STAT1 alpha, In vitro immunecomplex kinase assa y of PDGF beta receptor (PDGFR) or STAT1 alpha immunoprecipitates from lysates of mesangial cells treated with PDGF showed phosphorylation o f a 91- and an 185-kD protein. Incubation of lysates prepared from qui escent mesangial cells with purified PDGFR resulted in STAT1 alpha act ivation. Immunodepletion of Janus kinases from the cell lysate before incubation with the purified PDGFR showed no effect on STAT1 alpha act ivation. Moreover, lysates from mesangial cells treated with JAK2 inhi bitor, retained significant STAT1 alpha activity. To confirm that STAT 1 alpha is a substrate for PDGFR, STAT1 alpha protein was prepared by in vitro transcription and translation. The addition of purified PDGFR to the translated STAT1 alpha resulted in its phosphorylation, This i n vitro phosphorylated and activated protein a:lso forms a specific pr otein-DNA complex. Dimerization of the translated STAT1 alpha protein was also required for its DNA binding. Incubation of pure STAT1 alpha with autophosphorylated PDGFR resulted in physical association of the two proteins. These data indicate that activated PDGFR may be sufficie nt to tyrosine phosphorylate and thus directly activate STAT1 alpha.