A cytoplasmic serine protein kinase binds and may regulate the Fanconi anemia protein FANCA.
Fanconi anemia (FA) is an autosomal recessive disease with congenital anomalies, bone marrow failure, and susceptibility to leukemia. Patient cells show chromosome instability and hypersensitivity to DNA cross-linking agents. At least 8 complementation groups (A-G) have been identified and 6 FA genes (for subtypes A, C, D2, E, F, and G) have been cloned. Increasing evidence indicates that a protein complex assembly of multiple FA proteins, including FANCA and FANCG, plays a crucial role in the FA pathway. Previously, it was reported that FANCA was phosphorylated in lymphoblasts from normal controls, whereas the phosphorylation was defective in those derived from patients with FA of multiple complementation groups. The present study examined phosphorylation of FANCA ectopically expressed in FANCA(-) cells. Several patient-derived mutations abrogated in vivo phosphorylation of FANCA in this system, suggesting that FANCA phosphorylation is associated with its function. In vitro phosphorylation studies indicated that a physiologic protein kinase for FANCA (FANCA-PK) forms a complex with the substrate. Furthermore, at least a part of FANCA-PK as well as phosphorylated FANCA were included in the FANCA/FANCG complex. Thus, FANCA-PK appears to be another component of the FA protein complex and may regulate function of FANCA. FANCA-PK was characterized as a cytoplasmic serine kinase sensitive to wortmannin. Identification of the protein kinase is expected to elucidate regulatory mechanisms that control the FA pathway.
Yagasaki
H
H
Division of Genetic Diagnosis, Institute of Medical Science, University of Tokyo, Japan.
Adachi
D
D
Oda
T
T
Garcia-Higuera
I
I
Tetteh
N
N
D'Andrea
A D
AD
Futaki
M
M
Asano
S
S
Yamashita
T
T
D000730:Androstadienes / Q000494:pharmacology; D015153:Blotting, Western; D002461:Cell Line, Transformed; D003593:Cytoplasm / Q000201:enzymology*; D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005199:Fanconi Anemia / Q000201:enzymology*; D052217:Fanconi Anemia Complementation Group A Protein; D005347:Fibroblasts / Q000378:metabolism; D006098:Granulocytes / Q000378:metabolism; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D007163:Immunosorbent Techniques; D019169:Jurkat Cells / Q000378:metabolism; D008214:Lymphocytes / Q000378:metabolism; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D014162:Transfection; D000077191:Wortmannin
D000730:Androstadienes; D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; C498471:FANCA protein, human; D052217:Fanconi Anemia Complementation Group A Protein; D011506:Proteins; D017346:Protein Serine-Threonine Kinases; D000077191:Wortmannin