The transcription factor Pf-POU3F4 regulates expression of the matrix protein genes Aspein and Prismalin-14 in pearl oyster (Pinctada fucata)
The transcription factor Pf-POU3F4 regulates expression of the matrix protein genes Aspein and Prismalin-14 in pearl oyster (Pinctada fucata)
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转录因子 Pf-POU3F4 调节珍珠贝（Pinctada fucata）基质蛋白基因 Aspein 和 Prismalin-14 的表达
DOI：
10.1111/febs.13716
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发表时间：
2016-05-01
期刊：
影响因子：
5.4
通讯作者：
Zhang, Rongqing
中科院分区：
文献类型：
--
作者：
Gao, Jing;Chen, Yan;Zhang, Rongqing
基质蛋白在珍珠贝的贝壳形成中起着关键作用，但人们对这些蛋白是如何调控的知之甚少。在此，我们克隆并鉴定了POU结构域家族的两个成员：PF-POU2F1和PF-POU3F4。功能域分析表明，它们都有保守的POU和POUH结构域，这些结构域对转录因子的功能是重要的。Pf-POU2F1和Pf-POU3F4mRNAs在珍珠贝体内的组织分布呈现不同的表达模式，且Pf-POU3F4mRNAs在外套膜中的表达相对较高。利用基因组行走聚合酶链式反应技术克隆了基质蛋白基因Aspin和Prismarin-14的启动子。在HEK-293T细胞中检测到这些启动子具有较高的转录活性。在瞬时共转染实验中，PF-POU3F4以剂量依赖的方式显著上调Aspin和Prismarin-14基因的启动子活性。PF-POU3F4的结构完整性是其激活功能所必需的。Pf-POU3F4的激活需要Aspecin基因启动子-181~-77bp的一个区域和Prismarin-14基因启动子的-359~-337bp和-100~-73bp两个结合位点。电泳迁移率改变分析表明，PF-POU3F4直接结合了这些位点。Pf-POU3F4基因敲除导致Aspin和Prismarin-14基因表达下降。此外，Pf-POU3F4基因在五个发育阶段的表达水平与Aspecin和PrisMalin-14基因的表达水平相似。综上所述，这些结果表明转录因子PF-POU3F4调控珍珠牡蛎基质蛋白基因Aspin和Prismarin-14的表达。Pf-POU3F4的核苷酸序列数据可在GenBank数据库中找到，登录号为：Aspin基因启动子的核苷酸序列数据可在GenBank数据库中找到，登录号为。Prismarin-14基因启动子的核苷酸序列数据可在GenBank数据库中找到，登录号为。
Matrix proteins play key roles in shell formation in the pearl oyster, but little is known about how these proteins are regulated. Here, two POU domain family members, Pf-POU2F1 and Pf-POU3F4, were cloned and characterized. Functional domain analysis revealed that both them have conserved POUS and POUH domains; these domains are important for transcription factor function. The tissue distributions of Pf-POU2F1 and Pf-POU3F4 mRNAs in pearl oyster revealed different expression patterns, and the expression of Pf-POU3F4 mRNA was relatively high in the mantle. The promoters of the matrix protein genes Aspein and Prismalin-14 were cloned using genome-walking PCR. Relatively high transcriptional activities of these promoters were detected in HEK-293T cells. In transient co-transfection assays, Pf-POU3F4 greatly up-regulated the promoter activities of the Aspein and Prismalin-14 genes in a dose-dependent manner. Structural integrity of Pf-POU3F4 was essential for its activation function. One region of the Aspein gene promoter, -181 to -77bp, and two binding sites in the Prismalin-14 gene promoter, -359 to -337 bp and -100 to -73bp, were required for activation of Pf-POU3F4. An electrophoresis mobility shift assay demonstrated that Pf-POU3F4 directly bound these sites. Pf-POU3F4 knockdown led to a decrease in Aspein and Prismalin-14 gene expression. Furthermore, expression levels for the Pf-POU3F4 gene were similar to those of the Aspein and Prismalin-14 genes during five development stages. Taken together, these results suggest that the transcription factor Pf-POU3F4 regulates expression of the matrix protein genes Aspein and Prismalin-14 in pearl oyster.DatabaseThe nucleotide sequence data of Pf-POU2F1 is available in the GenBank databases under the accession number . The nucleotide sequence data of Pf-POU3F4 is available in the GenBank databases under the accession number . The nucleotide sequence data of Aspein gene promoter is available in the GenBank databases under the accession number . The nucleotide sequence data of Prismalin-14 gene promoter is available in the GenBank databases under the accession number .