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小鼠睾丸中一种新型Sox5转录本的鉴定。

Identification of a novel Sox5 transcript in mouse testis.

作者信息

Zheng Bangjin, Huang Chaoyang, Zhou Jian, Ye Lan

机构信息

State Key Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, China.

Department of Cardiology, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.

出版信息

Gene Expr Patterns. 2021 Sep;41:119197. doi: 10.1016/j.gep.2021.119197. Epub 2021 Jun 22.

Abstract

The transcription factor SOX5 is present in two distinct isoforms in both human and mouse, L-SOX5 and S-SOX5 (long and short isoforms of SOX5). Here, we identified and characterized a novel transcript of Sox5 (S-Sox5 variant) in mouse testis. eCLIP-based amplification of cDNA ends were performed to identify the potential Sox5 mRNA variant. This novel transcript shares a high similarity with the previously reported S-Sox5 in nucleotide sequence, but with a unique stretch of 5'UTR and an additional exon 9. Semi-quantitative PCR analysis revealed both S-Sox5 variant and S-Sox5 express specifically in mouse testis. Both transcripts increase significantly in mouse testis at postnatal day 21, when round spermatids appear. We further made a series of truncated Sox5 constructs and tagged them with eGFP in HeLa cells. In vitro transfection assay identified the N-terminus and the DNA-binding HMG domain are required for the nuclear localization of SOX5. Our results provides a basis for the future study to investigate the biological function of SOX5 in spermatogenesis.

摘要

转录因子SOX5在人类和小鼠中均以两种不同的异构体形式存在,即L-SOX5和S-SOX5(SOX5的长异构体和短异构体)。在此,我们在小鼠睾丸中鉴定并表征了一种新的Sox5转录本(S-Sox5变体)。通过基于eCLIP的cDNA末端扩增来鉴定潜在的Sox5 mRNA变体。这种新转录本在核苷酸序列上与先前报道的S-Sox5具有高度相似性,但具有独特的5'UTR延伸和一个额外的外显子9。半定量PCR分析显示,S-Sox5变体和S-Sox5均在小鼠睾丸中特异性表达。在出生后第21天圆形精子细胞出现时,这两种转录本在小鼠睾丸中的表达均显著增加。我们进一步构建了一系列截短的Sox5构建体,并在HeLa细胞中用eGFP标记它们。体外转染实验表明,SOX5的核定位需要N末端和DNA结合HMG结构域。我们的结果为未来研究SOX5在精子发生中的生物学功能提供了基础。

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