LincRNAs Regulate Atp2b2, Potentially Determining PMCA2 Quantity in Stereocilia

LincRNA 调节 Atp2b2，可能决定立体纤毛中 PMCA2 的数量

• 批准号: 8974974
• 负责人: BRUCE L TEMPEL
• 金额: $ 18.43万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8974974
• 关键词: 5' Untranslated Regions; Auditory; Auditory system; Binding; Brain Stem; Ca(2+)-Transporting ATPase; Calcium; Cell Line; Cell membrane; Cells; Chromosomes; Cochlea; Complex; Congenic Mice; Congenic Strain; DNA Sequence Alteration; Data; Down-Regulation; Exhibits; Exons; Gene Expression; Gene Targeting; Genes; Genetic Transcription; Genetic study; Haplotypes; Hearing; Homologous Gene; Human; Link; Mammalian Cell; Measures; Monitor; Mus; Mutation; Open Reading Frames; Organ of Corti; Outer Hair Cells; Peripheral; Phenotype; Play; Presbycusis; Process; Proteins; RNA; Regulation; Regulator Genes; Regulatory Element; Reporter; Research; Role; Sampling; Scientist; Sensory; Stereocilium; Testing; Transcript; Transcriptional Regulation; Translating; Untranslated RNA; Western Blotting; Work; deafness; hearing impairment; human CDH23 protein; inhibitor/antagonist; interest; link protein; overexpression; promoter; protein expression; public health relevance; research study; response; vector

 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.

 描述(由申请人提供)：C57BL/6J(B6)小鼠具有典型的年龄相关性听力损失(AHL)表型。最近的一项研究表明，这种损失只是由编码尖端连接蛋白CDH23的钙粘连蛋白23(CDH23)基因的突变部分造成的(Kane等人，2011年)。在小鼠和人类中，调节细胞内钙水平的质膜钙ATPase 2(PMCA2)蛋白的突变加剧了CDH23的突变(Noben-Trauth等人，1997和Schultz等人，2007)。这种互动是 可能是由于钙离子在维持CDH23结构完整性方面的必要性(Sotomayor等人，2010年)。编码PMCA2的B6Atp2b2基因没有突变。然而，两种谨慎的基因表达测量表明，与听力良好的菌株CBA/CAJ(CBA)相比，B6中Atp2b2转录下调。对Atp2b2的deafwaddler突变的研究表明，听觉系统对Atp2b2的微小变化高度敏感。这些小鼠表现出听力敏感性的变化，这可能与Atp2b2的调节、功能和表达的变化有关(McCullough和Tempel，2004；Watson和Tempel，2013)。所有这些证据表明，B6中Atp2b2的下调可能是这些小鼠年龄相关性听力损失表型的一个可能因素。由于Atp2b2蛋白编码区没有突变，但转录表达发生了变化，转录过程可能参与了Atp2b2在B6中的下调。坦佩尔实验室最近的实验证实，在小鼠Atp2b2基因的内含子区域中存在一个长的基因间非编码RNA(lincRNA-83)。表达研究表明，该基因在B6小鼠的脑干和耳蜗中存在调控错误。重要的是，lincRNAs正在成为附近基因转录调控的关键角色(Wang和Chang，2011)。这一建议中的最重要的假设是，B6中的Atp2b2错误调节导致了该菌株的AHL。为了更好地理解B6和CBA之间的表达差异，我们提出了两个目标：1)B6和CBA之间的表达差异程度，以及2)非编码RNA对Atp2b2基因的调控程度。