TENT-5 regulates the expression of male-specific genes in Caenorhabditis elegans

Polyadenylation is an important post-transcriptional process that governs mRNA stability and expression. Advancements in direct RNA sequencing in recent years have clarified many aspects of this intricate regulation, revealing the influence of various factors. Here, we used Nanopore Direct RNA Sequencing to investigate the association between genome-wide mRNA poly(A) tail profiles and sexual dimorphism in Caenorhabditis elegans. Our results demonstrate sex-dependent differences in both gene expression and poly(A) tail metabolism. Notably, we discovered that cytoplasmic poly(A) polymerase TENT-5 regulates multiple male-specific transcripts, predominantly encoding putative seminal fluid components with predicted extracellular localization. TENT-5 expression in male-specific tissues, such as seminal vesicle and vas deference, corroborates its functional significance. Intriguingly, despite extensive TENT-5-mediated polyadenylation of male-specific transcripts, males devoid of TENT-5 show no abnormalities in mating behavior, sperm morphology, or fertility. Our findings suggest that TENT-5 plays a role in regulating sex-related processes in males, although the physiological consequences remain to be fully elucidated. Overall design: We implemented Nanopore Direct RNA Sequencing to investigate sex-related differences in gene expression and poly(A) tail metabolism in C. elegans. We performed analysis of wild type (N2) hermaphrodites and males at the young adult stage. Additionally, we compared wild type (N2) males and tent-5(tm3504) males and showed that male-specific genes in C. elegans are regulated by TENT-5.

聚腺苷酸化（Polyadenylation）是调控信使RNA（mRNA）稳定性与表达的关键转录后过程。近年来，直接RNA测序（direct RNA sequencing）技术的突破阐明了这一复杂调控机制的诸多细节，揭示了多种调控因素的作用。本研究借助纳米孔（Nanopore）直接RNA测序技术，探究秀丽隐杆线虫（Caenorhabditis elegans）全基因组范围内mRNA的poly(A)尾谱与性别二态性之间的关联。研究结果表明，基因表达与poly(A)尾代谢均存在性别依赖性差异。值得注意的是，本研究发现细胞质聚腺苷酸聚合酶TENT-5可调控多种雄性特异性转录本，这些转录本主要编码推定的精液成分，且经预测具有细胞外定位特性。TENT-5在雄性特异性组织（如精囊与输精管）中的表达，进一步佐证了其功能重要性。有趣的是，尽管TENT-5可广泛介导雄性特异性转录本的聚腺苷酸化，但缺失TENT-5的雄性个体在交配行为、精子形态或生育能力上均未出现异常。本研究结果提示，TENT-5在雄性性别相关过程中发挥调控作用，但其具体生理效应仍有待全面阐明。总体实验设计：本研究采用纳米孔直接RNA测序技术，探究秀丽隐杆线虫年轻成虫阶段野生型（N2）雌雄同体与雄性个体在基因表达及poly(A)尾代谢方面的性别差异。我们对年轻成虫阶段的野生型（N2）雌雄同体与雄性个体进行了分析；此外，通过对比野生型（N2）雄性与tent-5(tm3504)突变雄性，证实秀丽隐杆线虫的雄性特异性基因受TENT-5调控。