Gonadotropin regulation of ankyrin-repeat and SOCS-box protein 9 (ASB9) in ovarian follicles and identification of binding partners

PloS One
Gabriel BenoitKalidou Ndiaye

Abstract

Ankyrin-repeat and SOCS-box protein 9 (ASB9) is a member of the large SOCS-box containing proteins family and acts as the specific substrate recognition component of E3 ubiquitin ligases in the process of ubiquitination and proteasomal degradation. We previously identified ASB9 as a differentially expressed gene in granulosa cells (GC) of bovine ovulatory follicles. This study aimed to further investigate ASB9 mRNA and protein regulation, identify binding partners in GC of bovine ovulatory follicles, and study its function. GC were obtained from small follicles (SF: 2-4 mm), dominant follicles at day 5 of the estrous cycle (DF), and ovulatory follicles, 24 hours following hCG injection (OF). Analyses by RT-PCR showed a 104-fold greater expression of ASB9 in GC of OF than in DF. Steady-state levels of ASB9 in follicular walls (granulosa and theca cells) analyzed at 0, 6, 12, 18 and 24 hours after hCG injection showed a significant induction of ASB9 expression at 12 and 18 hours, reaching a maximum induction of 10.2-fold at 24 hours post-hCG as compared to 0 hour. These results were confirmed in western blot analysis showing strongest ASB9 protein amounts in OF. Yeast two-hybrid screening of OF-cDNAs library resulted in the ident...Continue Reading

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Citations

Mar 12, 2020·Biology of Reproduction·Aly Warma, Kalidou Ndiaye
Feb 7, 2021·International Journal of Molecular Sciences·Aly WarmaKalidou Ndiaye
Sep 4, 2021·Molecular Reproduction and Development·Soma Nosratpour, Kalidou Ndiaye

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Methods Mentioned

BETA
two-hybrid
PCR
co-immunoprecipitation assay
transfection
co-immunoprecipitation
co-immunoprecipitations
ubiquitination

Software Mentioned

GraphPad prism
ProLabel
BLAST

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