Murine Spam1 mRNA: involvement of AU-rich elements in the 3'UTR and antisense RNA in its tight post-transcriptional regulation in spermatids.
Murine Spam1 mRNA: involvement of AU-rich elements in the 3'UTR and antisense RNA in its tight post-transcriptional regulation in spermatids.
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小鼠 Spam1 mRNA：3UTR 中富含 AU 的元件和反义 RNA 参与精子细胞的严格转录后调节。
DOI：
10.1002/mrd.20400
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发表时间：
2006
影响因子：
2.5
通讯作者：
Martin-Deleon,PatriciaA
中科院分区：
文献类型：
--
作者：
Zhang,Hong;Barnoski,BarryL;Sol-Church,Katia;Stabley,DeborahL;Martin-Deleon,PatriciaA
精子粘附分子1（SPAM 1）是透明质酸酶基因中最具代表性的基因，在睾丸中大量表达。我们试图通过转基因技术，使用BAC中的内源启动子或aSpam 1 cDNA转基因的异源鱼精蛋白1启动子来过表达小鼠Spam 1。尽管转基因拷贝数范围为2至15，并且表达了转基因转录物，但所有7名创始人的睾丸中RNA和蛋白质的过表达普遍失败。此外，三个转基因株系显示出BAC上存在的Spam 1和Hyal 5的RNA的适度下调或共抑制。我们提供的证据表明，两个协调的转录后调控过程可能参与过度表达的失败：丰富的内源性反义RNA和腺苷尿苷（Au）丰富的元素介导的RNA周转调节。我们证明，富含Au的元件（战神）在mRNA的3′UTR，众所周知的相互作用与反式作用蛋白质靶向RNA的稳定性，存在于Spam 1 RNA和特异性结合6睾丸细胞质蛋白。这些Au结合蛋白（AUBP）在肾脏中几乎不存在，肾脏中的转录物很少，并且显示与细胞骨架相互作用，调节mRNA周转。除了在RNAi途径中的作用外，反义RNA还可以通过与战神杂交并特异性沉默其功能来调节ARE介导的mRNA调节。这可能将Spam 1表达调控中的两个过程联系起来。我们假设testicularSpam1RNA在转录后受3′UTR中的顺式作用ARE调控，ARE识别AUBP并受反义转录调控。Reprod. Dev.© 2005 Wiley利斯公司
Sperm adhesion molecule1(SPAM1), the best characterized hyaluronidase gene, is abundantly expressed in the testis. We attempted to overexpress mouseSpam1via transgenesis using either the endogenous promoter in a BAC or a heterologousProtamine1promoter for aSpam1cDNA transgene. Although transgene‐copy numbers ranged from 2 to 15 and transgenic transcripts were expressed, there was a general failure of overexpression of the RNA and protein in the testis of all seven founders. Also, three transgenic lines showed a modest downregulation or co‐suppression of the RNA forSpam1andHyal5, present on the BAC. We provide evidence for the potential involvement of two co‐ordinating post‐transcriptional regulatory processes in the failure of overexpression: abundant endogenous antisense RNA and adenosine‐uridine (AU)‐rich element‐mediated regulation of RNA turnover. We demonstrate that AU‐rich elements (AREs) in the 3′UTR of mRNAs, well‐known to interact withtrans‐acting proteins to target the RNA for (in)stability, are present inSpam1RNA and specifically bind to six testicular cytoplasmic proteins. These AU‐binding proteins (AUBPs) were virtually absent from the kidney where transcripts are rare, and were shown to interact with the cytoskeleton, which modulates mRNA turnover. In addition to a role in the RNAi pathway, antisense RNA can also modulate ARE‐mediated regulation of mRNA by hybridizing to the AREs and specifically silencing their function. This potentially links the two processes in the regulation ofSpam1expression. We hypothesize that testicularSpam1RNA is regulated post‐transcriptionally bycis‐acting ARE(s) in the 3′UTR which recognize AUBPs and which are modulated by antisense transcripts.Mol. Reprod. Dev.© 2005 Wiley‐Liss, Inc.