Integrative Proteomic and Transcriptomic Analyses Reveal Multiple Post-transcriptional Regulatory Mechanisms of Mouse Spermatogenesis
Integrative Proteomic and Transcriptomic Analyses Reveal Multiple Post-transcriptional Regulatory Mechanisms of Mouse Spermatogenesis
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综合蛋白质组学和转录组学分析揭示了小鼠精子发生的多种转录后调节机制
DOI：
10.1074/mcp.m112.020123
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发表时间：
2013-05-01
影响因子：
7
通讯作者：
Han, Chunsheng
中科院分区：
文献类型：
--
作者：
Gan, Haiyun;Cai, Tanxi;Han, Chunsheng
哺乳动物精子发生包括多种细胞类型和生物学过程，是研究转录和转录后水平基因调控的理想模型。许多关键蛋白、miRNAs和piRNAs都参与了精子发生的转录后调控。然而，一个系统的方法来评估蛋白质和mRNA表达之间的关系还没有可用于研究转录后调控机制。在本研究中，我们使用iTRAQ为基础的定量蛋白质组学的方法来确定2008蛋白质在小鼠A型精原细胞，粗线期精母细胞，圆形精子细胞，和伸长精子细胞具有高的信心。在这些蛋白质中，1194个组成了四个动态变化的簇，它们反映了生殖细胞的有丝分裂扩增、减数分裂和减数分裂后的发育。我们根据有丝分裂/减数分裂过渡期和减数分裂/减数分裂后发育过渡期蛋白质水平相对于mRNA水平的变化确定了五种主要的调控机制，称为“仅转录本”、“转录本降解”、“翻译抑制”、“翻译去抑制”和“蛋白质降解”。我们发现转录后调节机制与piRNA和反义转录物的产生有关。我们的研究结果提供了一个有价值的库存蛋白在小鼠精子发生过程中，并有助于阐明哺乳动物精子发生中的基因表达的转录后调控机制。
Mammalian spermatogenesis consists of many cell types and biological processes and serves as an excellent model for studying gene regulation at transcriptional and post-transcriptional levels. Many key proteins, miRNAs, and perhaps piRNAs have been shown to be involved in post-transcriptional regulation of spermatogenesis. However, a systematic method for assessing the relationship between protein and mRNA expression has not been available for studying mechanisms of post-transcriptional regulation. In the present study, we used the iTRAQ-based quantitative proteomic approach to identify 2008 proteins in mouse type A spermatogonia, pachytene spermatocytes, round spermatids, and elongative spermatids with high confidence. Of these proteins, 1194 made up four dynamically changing clusters, which reflect the mitotic amplification, meiosis, and post-meiotic development of germ cells. We identified five major regulatory mechanisms termed "transcript only," "transcript degradation," "translation repression," "translation de-repression," and "protein degradation" based on changes in protein level relative to changes in mRNA level at the mitosis/meiosis transition and the meiosis/post-meiotic development transition. We found that post-transcriptional regulatory mechanisms are related to the generation of piRNAs and antisense transcripts. Our results provide a valuable inventory of proteins produced during mouse spermatogenesis and contribute to elucidating the mechanisms of the post-transcriptional regulation of gene expression in mammalian spermatogenesis.