ADAD2 functions in spermiogenesis and piRNA biogenesis in mice
ADAD2 functions in spermiogenesis and piRNA biogenesis in mice
复制标题
DOI：
10.1111/andr.13400
复制
发表时间：
2023-02-14
期刊：
影响因子：
4.5
通讯作者：
Ikawa，Masahito
中科院分区：
文献类型：
--
作者：
Lu，Yonggang;Nagamori，Ippei;Ikawa，Masahito
研究背景腺苷脱氨酶结构域2（Adenosine Deaminase domain containing 2，ADAD 2）是一种睾丸特异性蛋白，由一个双链RNA结合结构域和一个非催化性腺苷脱氨酶结构域组成。最近的一项研究表明，ADAD 2对于小鼠的雄性生殖是不可或缺的。目的本研究旨在探讨Adad 2突变小鼠雄性不育的原因，并了解ADAD 2的分子功能。材料与方法利用CRISPR/Cas9技术构建Adad 2纯合突变小鼠品系Adad 2-/-和Adad 2 Δ/Δ。Western blotting和免疫组化检测ADAD 2的表达和亚细胞定位。采用免疫共沉淀串联质谱法测定小鼠睾丸中ADAD 2相互作用蛋白。结果Adad 2-/-和Adad 2 Δ/Δ小鼠由于精子发生异常而表现出雄性特异性不育。ADAD 2与参与皮尔纳生物发生的多种RNA结合蛋白相互作用，包括MILI、MIWI、RNF 17和YTHDC 2。ADAD 2与RNF 17在精母细胞中共定位并形成新颗粒。ADAD 2的消融损害RNF 17颗粒的形成，减少簇衍生的粗线期piRNA的数量，并增加乒乓衍生的piRNAs.Discussion和conclusionIn与RNF 17和精母细胞中的其他RNA结合蛋白协作，ADAD 2直接或间接地在皮尔纳生物发生中起作用。
BackgroundAdenosine deaminase domain containing 2 (ADAD2) is a testis‐specific protein composed of a double‐stranded RNA binding domain and a non‐catalytic adenosine deaminase domain. A recent study showed that ADAD2 is indispensable for the male reproduction in mice. However, the detailed functions of ADAD2 remain elusive.ObjectivesThis study aimed to investigate the cause of male sterility inAdad2mutant mice and to understand the molecular functions of ADAD2.Materials and methodsAdad2homozygous mutant mouse lines,Adad2–/–andAdad2Δ/Δ, were generated by CRISPR/Cas9. Western blotting and immunohistochemistry were used to reveal the expression and subcellular localization of ADAD2. Co‐immunoprecipitation tandem mass spectrometry was employed to determine the ADAD2‐interacting proteins in mouse testes. RNA‐sequencing analyses were carried out to analyze the transcriptome and PIWI‐interacting RNA (piRNA) populations in wildtype andAdad2mutant testes.ResultsAdad2–/–andAdad2Δ/Δmice exhibit male‐specific sterility because of abnormal spermiogenesis. ADAD2 interacts with multiple RNA‐binding proteins involved in piRNA biogenesis, including MILI, MIWI, RNF17, and YTHDC2. ADAD2 co‐localizes and forms novel granules with RNF17 in spermatocytes. Ablation of ADAD2 impairs the formation of RNF17 granules, decreases the number of cluster‐derived pachytene piRNAs, and increases expression of ping‐pong‐derived piRNAs.Discussion and conclusionIn collaboration with RNF17 and other RNA‐binding proteins in spermatocytes, ADAD2 directly or indirectly functions in piRNA biogenesis.