Src family tyrosine kinases phosphorylate Flt3 on juxtamembrane tyrosines and interfere with receptor maturation in a kinase-dependent manner.
The receptor tyrosine kinase (RTK) Flt3 is expressed in early hematopoietic progenitor cells and stimulates their growth. Due to frequent mutations in the Flt3 gene in patients with acute myeloid leukemia (AML), Flt3 is regarded as a potential therapeutic target, but the underlying mechanisms are still poorly understood. Therefore, we investigated interactions of Flt3 and some Src family tyrosine kinases (SFKs), which are expressed predominantly or exclusively in hematopoietic cells and known to be involved in signal transduction by various RTKs. Employing sets of wt and mutant Flt3 and Hck, we analyzed protein binding as well as Flt3 phosphorylation and maturation in HEK-293 cells cotransfected with expression constructs encoding both binding partners. Kinase-inactive Hck-K269R was recruited to phosphotyrosine residues located in the juxtamembrane (JM) region of activated Flt3 via its SH2 domain. Several of the JM domain tyrosines were phophorylated by Hck and other SFKs. As apparent from the distribution of mature and hypoglycosylated Flt3, SFKs interfered with Flt3 maturation in a kinase-dependent manner. Together, these findings show a complex role of SFKs in Flt3 signaling and reveal a new function of SFKs in the maturation of RTKs.
Clinical Cooperation Group Gene Therapy, GSF-National Research Center for Environment and Health, Marchioninistr 25, 81377, Munich, Germany.
Olga
O
Mitina
Markus
M
Warmuth
Günter
G
Krause
Michael
M
Hallek
Axel
A
Obermeier
D048708:Cell Growth Processes / Q000502:physiology*; D002460:Cell Line; D006410:Hematopoiesis / Q000502:physiology; D006801:Humans; D051572:Proto-Oncogene Proteins c-hck / Q000378:metabolism; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism*; D015398:Signal Transduction; D014162:Transfection; D051941:fms-Like Tyrosine Kinase 3 / Q000378:metabolism*; D019061:src-Family Kinases / Q000378:metabolism*
C497970:FLT3 protein, human; D020794:Receptor Protein-Tyrosine Kinases; D051941:fms-Like Tyrosine Kinase 3; C496631:HCK protein, human; D051572:Proto-Oncogene Proteins c-hck; D019061:src-Family Kinases