The Study Of Lysine Specific Demethylase1(LSD1) Interact With Transcription Factor GATA2Regulate Hematopoiesis Differentiation

Lysine-specific demethylase1（LSD1） was recently identified as the first histonelysine demethylase that specifically demethylates histone H3at K4and K9. LSD1canonly demethylate monomethylated and dimethylated H3K4and H3K9because thereaction lead by LSD1need to form an imine intermediate. LSD1express itsepigentics function regulating chromatin structure through demethylation histoneprotein. Recent study shows LSD1can regulate the development of leukemia, but themechanism is still unknown.We found that LSD1expressed in most of leukemia cells. Following celldifferentiation, the expression of LSD1is decreased. While, there are studies foundthe significant hematopoiesis transcription factor GATA2also down regulate itsexpression. To test whether LSD1regulate hematopoietic cell differentiation thoughinteract with GATA2, we design the next experiment.We confirm LSD1interact with GATA2in murine erythroleukemia cell andhuman erythroleukemia cell by immunoprecipitation. To test whether LSD1directinteract with GATA2and the interaction domain, we construct, express and purifydifferent GST fusion protein with different domain of LSD1(GST-LSD1^N-term、GST-LSD1^N-term+SWIRM、GST-LSD1^SWIRM、GST-LSD1^SWIRM+AO、GST-LSD1^AO). Thenusing GST Pull-down assay finds LSD1can directly interact with GATA2through itsSWIRM domain. Meanwhile the N-term of LSD1affects the interaction betweenSWIRM domain and GATA2. Further experiment we test the function of LSD1inhematopoietic cell differentiation by using RNAi to knock down LSD1expression.We find that the transcription of GATA2is obvious increased in LSD1kd MEL cells,while the expression of GATA1is decreased. In our research, we also using celldifferentiation assay prove that LSD1kd inhibits Mel cell differentiation.In conclusion, our experiment certificates LSD1directly interacts with GATA2through SWIRM domain to inhibit the transcription of GATA2in the hematopoieticdifferentiation. By the interaction with GATA2, LSD1participate in GATA Switch andaffect hematopoietic differentiation. Our program makes the foundation of the study of hematopoiesis differentiation process, find new substrate for leukemia treatment.