Key Laboratory of Epigenetics and Oncology, the Research Center for Preclinical Medicine, Southwest Medical University, 3-319, Zhongshan Rd, Luzhou, 646000, Sichuan, China.
Department of Gynecology and Obstetrics, Guangzhou Women and Children's Hospital, Guangzhou, Guangdong, China.
Mol Biol Rep. 2019 Aug;46(4):3827-3834. doi: 10.1007/s11033-019-04825-4. Epub 2019 Apr 20.
This study aimed to clone and characterize novel isoforms of the human SPATA3 gene. The isoforms of SPATA3 gene was cloned into pGMT vector using human testis cDNA as template, and Sanger sequencing was performed. Their characterizations and tissue-specific expression profiles were analyzed. The two novel isoforms were successfully cloned and deposited into GenBank as MG029442 (AYP71042) and MG029443 (AYP71043) respectively. Isoforms SPATA3-I1 and SPATA3-I2 were found with higher identity, where only 7 amino acids missed at N-terminus in SPATA3-I2, whereas SPATA3-I3 and SPATA3-I4 had more C-terminus deletion but in SPATA3-I3 no amino acid missed at N-terminus. Importantly, we found the characterization of QQPSPESTP domain with two repeats for isoforms SPATA3-I1 and SPATA3-I4, whereas three repeats for isoforms SPATA3-I1 and SPATA3-I2. The SPATA3 family of genes is orthologous conserved; the similar core PEST domain was also revealed with variable repeats, indicating that this domain may pay roles in the spermatogenesis and male development differently. Furthermore, RNA-seq data indicated that the SPATA3 gene is only expressed in testis. This further suggests that SPATA3 plays potential roles only in male development, spermatogenesis or spermatogenesis cell apoptosis. Thus, in this study we cloned the two novel isoforms of SPATA3, SPATA3-I3 and SPATA3-I4, and found interesting characteristic PEST domain (QQPSPESTP) conserved in different isoforms as well as in different species. SPATA3 is an essential gene and may functions in male reproductive system, specifically in spermatogenesis.
本研究旨在克隆和鉴定人类 SPATA3 基因的新型异构体。使用人睾丸 cDNA 作为模板，将 SPATA3 基因的异构体克隆到 pGMT 载体中，并进行 Sanger 测序。分析它们的特征和组织特异性表达谱。成功克隆了两种新型异构体，并分别将其提交到 GenBank 作为 MG029442（AYP71042）和 MG029443（AYP71043）。发现 SPATA3-I1 和 SPATA3-I2 具有更高的同源性，其中 SPATA3-I2 的 N 端仅缺失 7 个氨基酸，而 SPATA3-I3 和 SPATA3-I4 的 C 端缺失更多，但 SPATA3-I3 的 N 端没有氨基酸缺失。重要的是，我们发现具有两个重复的 QQPSPESTP 结构域的 SPATA3-I1 和 SPATA3-I4 异构体，而具有三个重复的 SPATA3-I1 和 SPATA3-I2 异构体。SPATA3 基因家族是直系同源保守的；也揭示了具有可变重复的类似 PEST 核心结构域，表明该结构域可能在精子发生和男性发育中发挥不同的作用。此外，RNA-seq 数据表明 SPATA3 基因仅在睾丸中表达。这进一步表明 SPATA3 仅在男性发育、精子发生或精母细胞凋亡中发挥潜在作用。因此，在这项研究中，我们克隆了 SPATA3 的两种新型异构体 SPATA3-I3 和 SPATA3-I4，并发现了有趣的特征 PEST 结构域（QQPSPESTP）在不同异构体以及不同物种中保守。SPATA3 是一个必需基因，可能在男性生殖系统中发挥作用，特别是在精子发生中。
