Regulation of Ankyrin‐repeat and SOCS‐box protein 9 (ASB9) in ovarian follicles and identification of binding partners
Bibliographic record
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. ASB9 is known to interact with creatine kinase B and negatively regulates cell growth. 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 and to identify its binding partners in GC of bovine ovulatory follicles. 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). RT‐PCR analyses showed a 104‐fold induction of ASB9 expression in GC of OF as compared to DF (P < 0.0001). Steady‐state mRNA levels of ASB9 in follicular walls (granulosa and theca cells) analyzed at 0, 6, 12, 18 and 24 h after hCG injection showed a significant induction of ASB9 expression at 12 and 18 h (P<0.001), reaching a maximum induction at 24 h post‐hCG (P<0.0001) as compared to 0 h. These results were confirmed in western blot analysis showing highest ASB9 protein amounts in OF. Yeast two‐hybrid screening of OF‐cDNAs library resulted in the identification of five potential ASB9 binding partners in GC, confirmed by in vitro co‐immunoprecipitation analyses. Overall, these results support a physiologically relevant role of ASB9 in the ovulation process, and provide, for the first time, insights into the regulation and mode of action of ASB9 in GC. Support or Funding Information This work was supported in part by internal funds from Université de Montréal to KN and by a Discovery Grant from Natural Sciences and Engineering Research Council of Canada (NSERC) to JGL.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.001 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".