A potential role of Gsdf-eEF1 complex in inhibiting germ cell proliferation: A protein interaction analysis in medaka (Oryzias latipes) from a proteomics perspective

ABSTRACT: Gonadal soma-derived factor (gsdf) has been demonstrated as essential for testicular differentiation in medaka (Oryzias latipes). To understand the protein dynamics of Gsdf in spermatogenesis regulation, we used a His-tag “pull-down” assay coupled with shotgun LC-MS/MS to identify a group of potential interaction partners for Gsdf, which included cytoplasmic dynein light chain 2, eukaryotic polypeptide elongation factor 1 alpha (eEF1), and actin filaments in mature medaka testis. Alike the interaction with TGF-dynein critical for spermatogonial division in Drosophila melanogaster, the physical interaction of Gsdf & dynein and Gsdf & eEF1 were identified through a yeast 2-hybrid (Y2H) screening of an adult testis cDNA library using Gsdf as bait, and were verified by a paired Y2H assay. Co-immunoprecipitation of Gsdf and eEF1was defined in adult testes, as supporting the requirement of Gsdf and eEF1 interaction in testis development. Proteomics analysis and ultrastrutural observation showed that Gsdf deficiency activated eEF1-mediated protein synthesis and ribosome biogenesis, which in turn led to the differentiation of undifferentiated germ cell. Thus, our results provide a framework and new insight into the coordination of Gsdf (TGF and eEF1complex in the basic processes of germ cell proliferation, and transcriptional and translational control of sexual RNA, which may be fundamentally conserved across phyla during sex differentiation.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Oryzias Latipes (japanese Rice Fish) (japanese Killifish)

SUBMITTER: Guijun Guan  

LAB HEAD: Guijun Guan

PROVIDER: PXD022153 | Pride | 2020-12-09

REPOSITORIES: Pride