New Hematopoietic A Preliminary Study Of The Mechanism Of The Transcriptional Regulator Edag Role

Erythroid differentiation-associated gene (EDAG), which is homologous to mouse hemogen and rat RP59, is considered to be a human haematopoiesis-specific gene and mapped to chromosome 9q22. It was identified by representational difference analysis (RDA) between fetal and adult liver. We acquired the whole cDNA of 2166 base pairs, which encodes 484 amino acides. Studies indicate that EDAG gene is highly expressed in embryonic and haematopoietic tissues and overexpression of EDAG in NIH3T3 cells results in malignant transformation. EDAG gene is relative to maintaining undifferentiation of cells. Overexpression of EDAG in pro-B Ba/F3 cells prolongs survival and increases the expression of c-Myc, Bcl-2 and Bcl-xl in the absence of interleukin-3(IL-3) by enhancing the transcriptional activity of nuclear factor-kappa B. Studies of transgenetic mouse show that overexpression of EDAG in haematopoietic cells of mouse leads to disturbance of proliferation and differentiation of haemopoietic stem cell (HSC).Taken together, these data suggest that the EDAG gene might play an important role in haematopoietic development and neoplasm.Nowadays, the mechanisms of EDAG regulating proliferation, differentiation and apoptosis of haematopoietic cells remain largely unknown. Bioinformatics analysis suggests that N-terminus of EDAG protein contains a typical coiled-coiled domain. Coiled-coiled domain is highly conserved in monomer protein and mediate interactions of proteins .So EDAG maybe play an important role in haematogenesis through interaction with other important proteins.We prepare the polyclonal antibodies of EDAG and THAP11, detect the expression of EDAG and THAP11 at protein levels, which show the similar expression between EDAG and THAP11, and test the interaction of EDAG and THAP11 by co-localization and co-immunoprecipitation. Preliminary study shows that THAP11 can enhance the transcriptional activity of EDAG. We detect the presence of EDAG-GFP protein in the nucleus, which identified by Western blot furthermore. We discover that HSP70 is one of the EDAG interaction proteins by...