LincRNAs Regulate Atp2b2, Potentially Determining PMCA2 Quantity in Stereocilia

 描述（由申请人提供）：C57 BL/6 J（B6）小鼠具有良好表征的年龄相关性听力损失（阿勒）表型。最近的研究表明，这种损失仅部分由编码尖端连接蛋白CDH 23的钙粘蛋白23（Cdh 23）基因中的突变引起（Kane等人，2011年）。在小鼠和人类中，Cdh 23的突变被调节细胞内Ca 2+水平的质膜Ca 2 + ATP酶2（PMCA 2）蛋白的突变加剧（Noben-Trauth et al.，1997和Schultz等人，2007年）。这种相互作用 这可能是由于Ca 2+在维持CDH 23的结构完整性中的必要性（Sotomayor等，2010年）。在编码PMCA 2的B6 Atp 2b 2基因中没有突变。然而，基因表达的两个谨慎的措施表明，有下调Atp 2b 2转录B6相比，良好的听力菌株CBA/CaJ（CBA）。对Atp 2b 2中突变的研究表明，听觉系统对Atp 2b 2的微小变化高度敏感。这些小鼠表现出与Atp 2b 2的调节、功能和表达变化相关的听力灵敏度变化（McCullough和Tempel，2004;沃森和Tempel，2013）。所有这些证据表明，B6中Atp 2b 2的下调可能是这些小鼠中年龄相关性听力损失表型的贡献者。由于Atp 2b 2的蛋白编码区没有突变，但转录表达发生了变化，因此转录过程可能参与了B6中Atp 2b 2的下调。Tempel实验室最近的实验已经证实了在小鼠Atp 2b 2基因的内含子区域中存在长的基因间非编码RNA（lincRNA-83）。表达研究表明，该基因在B6小鼠的脑干和耳蜗中失调。重要的是，lincRNA正在成为附近基因转录调控的关键参与者（Wang和Chang，2011）。该建议中的过度假设是B6中Atp 2b 2的失调导致了该菌株中的阿勒。我们提出了两个目的，以更好地了解：1）B6和CBA之间的表达差异程度，以及2）非编码RNA调节Atp 2b 2基因的程度。

 DESCRIPTION (provided by applicant): C57BL/6J (B6) mice have a well-characterized age-related hearing loss (AHL) phenotype. A recent study has shown that this loss is only partially caused by mutations in the Cadherin 23 (Cdh23) gene which encodes the tip link protein CDH23 (Kane et al., 2011). In mice and in humans, mutations in Cdh23 are exacerbated by mutations in the plasma membrane Ca2+ ATPase 2 (PMCA2) protein, which regulates intracellular Ca2+ levels (Noben-Trauth et al., 1997 and Schultz et al., 2007). This interaction is likely due to the necessity of Ca2+ in maintaining the structural integrity of CDH23 (Sotomayor et al., 2010). There are no mutations in the B6 Atp2b2 gene, which encodes PMCA2. However, two discreet measures of gene expression show that there is down-regulation of Atp2b2 transcript in B6 compared to the good-hearing strain CBA/CaJ (CBA). Studies of the deafwaddler mutations in Atp2b2 demonstrate that the auditory system is highly sensitive to small changes in Atp2b2. These mice exhibit changes in hearing sensitivity that can be correlated to changes in regulation, function and expression of Atp2b2 (McCullough and Tempel, 2004; Watson and Tempel, 2013). All of this evidence suggests that the down-regulation of Atp2b2 in B6 is a likely contributor to the age-related hearing loss phenotype in these mice. As there are no mutations in the protein coding region of Atp2b2 but changes in transcript expression, transcriptional processes are likely involved in the down-regulation of Atp2b2 in B6. Recent experiments in the Tempel lab have confirmed the presence of a long intergenic non-coding RNA (lincRNA-83) that is in the intronic regions of the mouse Atp2b2 gene. Expression studies indicate that this gene is misregulated in the brainstem and the cochlea of B6 mice. Importantly, lincRNAs are emerging as key players in transcriptional regulation of nearby genes (Wang and Chang, 2011). The over-arching hypothesis in this proposal is that Atp2b2 misregulation in B6 contributes to AHL in this strain. We propose two aims to better understand the: 1) degree of expression differences between B6 and CBA, and 2) the extent to which non-coding RNAs regulate the Atp2b2 gene.