Molecular Mechanism Of Asz1to Promote PGC Formation From Murine Embryonic Stem Cell

All germ cells including sperms and eggs are derived from primordial germ cell in mammals. It is thus very important to understand the differentiation process and mechanism of PGC formation during germ cell development,but the number of Primordial Germ Cell from early embryo is limited. In recent years, stem cell research developed rapidly and this provides a very useful tool for the research of developmental biology. The embryonic stem cell (ESCs) can differentiate into almost any cell types include germ cells. We try to explore the molecular mechanism of PGC differentiation from ESCs and screen many genes relevant to reproduction by SSEA1staining and AP staining. We found that ASZ1can promote the process of differentiation from ESCs to PGCs.In this study, we found that asz1was localized in the mitochondria, asz1contained four ankyrin (ANK) repeats, a sterile-a motif (SAM) and a putative basic leucine zipper (bZIP) domain. The c-terminal signal peptide was very important for its localization in mitochodria.ASZ1lost its function when it changed its localization.We also identified DAZL as the interaction protein of ASZ1by mass spectrum analysis.We further confirmed DAZL can bind to ASZ1by western-blot and co-IP experiments. Also We found that DAZL bind to ASZ1through its SAM domain. We confirmed that ASZ1and DAZL colocalizised by bimolecular fluorescence complementation.We found that ASZ1lost its function when DAZ1was knockdowning. We also observed that overexpression of ASZ1and DAZL could synergistically promotr the PGC formation.In conclusion, that ASZ1can promote PGC formation and asz1was localized in the mitochondria. In addition, ASZ1can bind to DAZL. ASZ1will lose its function upon the change of its subcellular localization or the knowdowning of DAZL.