[Construction of FANCA mutant protein from Fanconi anemia patient and analysis of its function].
OBJECTIVE
To study FANCA protein expression in Fanconi anemia patient's (FA) cells and explore its function.


METHODS
FANCA protein expression was analyzed in 3 lymphoblast cell lines derived from 3 cases of type A FA (FA-A) patients using Western blot. Nucleus and cytoplasm localization of FANCA protein was analyzed in one case of FA-A which contained a truncated FANCA (exon 5 deletion). The FANCA mutant was constructed from the same patient and its interaction with FANCG was evaluated by mammalian two-hybrid (M2H) assay.


RESULTS
FANCA protein was not detected in the 3 FA-A patients by rabbit anti-human MoAb, but a truncated FANCA protein was detected in 1 of them by mouse anti-human MoAb. The truncated FANCA could not transport from cytoplasm into nucleus. The disease-associated FANCA mutant was defective in binding to FANCG in M2H system.


CONCLUSIONS
FANCA proteins are defective in the 3 FA-A patients. Disfunction of disease-associated FANCA mutant proved to be the pathogenic mutations in FANCA gene. Exon 5 of FANCA gene was involved in the interaction between FANCA and FANCG.
Chen
Fei
F
Department of Hematology, Zhongnan Hospital of Wuhan University, Wuhan 430071, China.
Zhang
Ke-Jian
KJ
Zuo
Xue-Lan
XL
Zeng
Xian-Chang
XC
D002460:Cell Line; D002648:Child; D005091:Exons; D005199:Fanconi Anemia / Q000235:genetics* / Q000378:metabolism; D052217:Fanconi Anemia Complementation Group A Protein / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism; D008297:Male; D009154:Mutation
D052217:Fanconi Anemia Complementation Group A Protein