Dataset Information

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

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Publications

A Potential Role for the Gsdf-eEF1α Complex in Inhibiting Germ Cell Proliferation: A Protein-Interaction Analysis in Medaka (Oryzias latipes) From a Proteomics Perspective.

Zhang Xinting X Chang Yuyang Y Zhai Wanying W Qian Feng F Zhang Yingqing Y Xu Shumei S Guo Haiyan H Wang Siyu S Hu Ruiqin R Zhong Xiaozhu X Zhao Xiaomiao X Chen Liangbiao L Guan Guijun G

Molecular & cellular proteomics : MCP 20210107

Gonadal soma-derived factor (gsdf) has been demonstrated to be 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 interacting partners for Gsdf, which included cytoplasmic dynein light chain 2, eukaryotic polypeptide elongation factor 1 alpha (eEF1α), and actin filaments in the mature medaka testis. As for ...[more]

PMID: 33293461

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Project description:Male song sparrows (Melospiza melodia) are territorial year-round; however, neuroendocrine responses to simulated territorial intrusion (STI) differ between breeding (spring) and non-breeding seasons (autumn). In spring, exposure to STI leads to increases in luteinizing hormone and testosterone, but not in autumn. These observations suggest that there are fundamental differences in the mechanisms driving neuroendocrine responses to STI between seasons. Microarrays, spotted with EST cDNA clones of zebra finch, were used to explore gene expression profiles in the hypothalamus after territorial aggression in two different seasons. Free-living territorial male song sparrows were exposed to either conspecific or heterospecific (control) males in an STI in spring and autumn. Behavioral data were recorded, whole hypothalami were collected, and microarray hybridizations were performed. Quantitative PCR was performed for validation. Our results show 262 cDNAs were differentially expressed between spring and autumn in the control birds. There were 173 cDNAs significantly affected by STI in autumn; however, only 67 were significantly affected by STI in spring. There were 88 cDNAs that showed significant interactions in both season and STI. Results suggest that STI drives differential genomic responses in the hypothalamus in the spring vs. autumn. The number of cDNAs differentially expressed in relation to season was greater than in relation to social interactions, suggesting major underlying seasonal effects in the hypothalamus which may determine the differential response upon social interaction. Functional pathway analyses implicated genes that regulate thyroid hormone action and neuroplasticity as targets of this neuroendocrine regulation. For condition experiment, SC (spring control), AC (autumn control), SE (spring STI), AE (autumn STI) Biological replicates: 7 in SC, 8 in AC, SE, AE. One Melospiza melodia subject and one universal SoNG reference (pooled Taeniopygia guttata brain) per array.

Dataset Information

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

Publications

A Potential Role for the Gsdf-eEF1α Complex in Inhibiting Germ Cell Proliferation: A Protein-Interaction Analysis in Medaka (Oryzias latipes) From a Proteomics Perspective.

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