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Molecular Mechanism of HINTW (HINT Gene in W-Chromosome)-Mediated UBE2I (Ubiquitin Conjugating Enzyme E2 I) Interaction in Female Differentiation of Chicken Embryos

文献类型： 外文期刊

作者： Sun, Changhua 1 ; Song, Jiuzhou 5 ; Ali, Malik Ahsan 2 ; Sun, Hongyan 2 ; Niu, Yingjie 2 ; Zuo, Qisheng 2 ; Han, Wei 7 ; Li, Bichun 2 ; Jin, Kai 2 ;

作者机构： 1.Yangzhou Polytech Coll, Coll Biol & Chem Engn, Yangzhou 225009, Peoples R China

2.Yangzhou Univ, Joint Int Res Lab Agr & Agriprod Safety, Minist Educ China, Yangzhou 225009, Peoples R China

3.Yangzhou Univ, Coll Anim Sci & Technol, Key Lab Anim Breeding Reprod & Mol Design Jiangsu, Yangzhou 225009, Peoples R China

4.Yangzhou Univ, Inst Agr Sci & Technol Dev, Yangzhou 225009, Peoples R China

5.Univ Maryland, Dept Anim & Avian Sci, College Pk, MD 20742 USA

6.Yangzhou Univ, Coll Biosci & Biotechnol, Yangzhou 225009, Peoples R China

7.Chinese Acad Agr Sci, Poultry Inst, Jiangsu Inst Poultry Sci, Yangzhou 225003, Peoples R China

8.Jiangsu Univ Sci & Technol, Coll Biotechnol, Zhenjiang 212100, Peoples R China

关键词： HINTW; UBE2I; female differentiation; chicken embryo; pull-down assay

期刊名称：GENES （ 影响因子：2.8； 五年影响因子：3.2 ）

ISSN：

年卷期： 2025 年 16 卷 4 期

页码：

收录情况： SCI

摘要： Background/Objective: It has been established that HINTW plays a pivotal role in the female differentiation of chickens; nevertheless, the underlying molecular mechanism remains to be fully elucidated. Method: To investigate the role of HINTW in avian sex determination, a prokaryotic expression vector containing its key structural domain was constructed, and its in vitro expression was achieved. Pull-down assays were performed to capture interacting proteins from male and female gonadal tissues, followed by a silver staining analysis to compare interaction profiles between ovaries and testes. Mass spectrometry was utilized to identify differentially bound proteins. Additionally, functional characterization and co-immunoprecipitation (Co-IP) assays were conducted to validate the interaction between HINTW and its candidate binding partner. Result: A total of 1590 differentially bound proteins were identified between ovarian and testicular tissues. Functional analysis and Co-IP assays confirmed a specific interaction between HINTW and UBE2I in the ovary, suggesting that HINTW may facilitate female differentiation in chicken embryos through its interaction with UBE2I. Conclusions: This study provides novel insights into the molecular mechanisms underlying HINTW-mediated female differentiation in chickens and contributes to a deeper understanding of avian sex determination.

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