Substrate specificities and identification of a putative binding site for PI3K in the carboxy tail of the murine Flt3 receptor tyrosine kinase.
Flt3 is a receptor tyrosine kinase (RTK) structurally related to the CSF-1R encoded by the c-fms locus, Kit and the PDGFR which is restricted in its expression to hematopoietic precursor populations and several distinct cell types within the central nervous system. Although the ligand for Flt3 has recently been identified, the developmental function of Flt3 within these tissues has not yet been described. In order to examine the signalling properties of this receptor, we previously constructed a chimeric molecule containing the extracellular domain of CSF-1R fused to the transmembrane and cytoplasmic domain of mouse Flt3 (FF3). The ability of the FF3 to directly associate with or tyrosine phosphorylate specific cytoplasmic signalling molecules in vivo was examined. GAP, Vav, Shc, and to a lesser extent PLC gamma become tyrosine-phosphorylated but no in vivo association with the receptor was detectable. FF3 associates with PI3K activity and the SH2 domains of p85 and Grb-2. Phosphopeptide competition experiments suggest that the PI3K binding site is located outside of the kinase insert in the carboxy tail of the receptor.
Rottapel
R
R
Molecular Hematology Laboratory, Unite 119 INSERM, Marseille, France.
Turck
C W
CW
Casteran
N
N
Liu
X
X
Birnbaum
D
D
Pawson
T
T
Dubreuil
P
P
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D020690:GTPase-Activating Proteins; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D019869:Phosphatidylinositol 3-Kinases; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020794:Receptor Protein-Tyrosine Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016186:Receptor, Macrophage Colony-Stimulating Factor / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013379:Substrate Specificity; D010738:Type C Phospholipases / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism; D051941:fms-Like Tyrosine Kinase 3
D020690:GTPase-Activating Proteins; D011506:Proteins; D011518:Proto-Oncogene Proteins; D011993:Recombinant Fusion Proteins; D014443:Tyrosine; D016173:Macrophage Colony-Stimulating Factor; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor); C497971:Flt3 protein, mouse; D020794:Receptor Protein-Tyrosine Kinases; D016186:Receptor, Macrophage Colony-Stimulating Factor; D051941:fms-Like Tyrosine Kinase 3; D010738:Type C Phospholipases