Snyder Elizabeth M, Licht Konstantin, Braun Robert E
The Jackson Laboratory, Bar Harbor, Maine, USA.
Department of Chromosome Biology, Max F. Perutz Laboratories, University of Vienna, Vienna, Austria.
Biol Reprod. 2017 Jan 1;96(1):244-253. doi: 10.1095/biolreprod.116.145151.
Adenosine to inosine (A-to-I) RNA editing occurs in a wide range of tissues and cell types and can be catalyzed by one of the two adenosine deaminase acting on double-stranded RNA enzymes, ADAR and ADARB1. Editing can impact both coding and noncoding regions of RNA, and in higher organisms has been proposed to function in adaptive evolution. Neither the prevalence of A-to-I editing nor the role of either ADAR or ADARB1 has been examined in the context of germ cell development in mammals. Computational analysis of whole testis and cell-type specific RNA-sequencing data followed by molecular confirmation demonstrated that A-to-I RNA editing occurs in both the germ line and in somatic Sertoli cells in two targets, Cog3 and Rpa1. Expression analysis demonstrated both Adar and Adarb1 were expressed in both Sertoli cells and in a cell-type dependent manner during germ cell development. Conditional ablation of Adar did not impact testicular RNA editing in either germ cells or Sertoli cells. Additionally, Adar ablation in either cell type did not have gross impacts on germ cell development or male fertility. In contrast, global Adarb1 knockout animals demonstrated a complete loss of A-to-I RNA editing in spite of normal germ cell development. Taken together, these observations demonstrate ADARB1 mediates A-to-I RNA editing in the testis and these editing events are dispensable for male fertility in an inbred mouse strain in the lab.
腺苷到次黄苷(A到I)的RNA编辑发生在广泛的组织和细胞类型中,可由作用于双链RNA的两种腺苷脱氨酶之一ADAR和ADARB1催化。编辑可影响RNA的编码区和非编码区,在高等生物中,有人提出其在适应性进化中发挥作用。在哺乳动物生殖细胞发育的背景下,尚未研究A到I编辑的普遍性以及ADAR或ADARB1的作用。通过对全睾丸和细胞类型特异性RNA测序数据进行计算分析,随后进行分子确认,结果表明在两个靶点Cog3和Rpa1中,A到I的RNA编辑发生在生殖系和体细胞支持细胞中。表达分析表明,在生殖细胞发育过程中,Adar和Adarb1在支持细胞中均有表达,且呈细胞类型依赖性。Adar的条件性敲除对生殖细胞或支持细胞中的睾丸RNA编辑均无影响。此外,在任何一种细胞类型中敲除Adar对生殖细胞发育或雄性生育能力均无明显影响。相比之下,尽管生殖细胞发育正常,但全局敲除Adarb1的动物显示A到I的RNA编辑完全丧失。综上所述,这些观察结果表明ADARB1介导睾丸中的A到I RNA编辑,并且在实验室近交小鼠品系中,这些编辑事件对于雄性生育能力是可有可无的。