The Role of Transcription Factor Isoforms in Eye and Neural Crest Development

转录因子亚型在眼睛和神经嵴发育中的作用

• 批准号: 7594712
• 负责人: HEINZ ARNHEITER
• 金额: $ 82.07万
• 依托单位: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7594712
• 关键词: Affect; Alternative Splicing; Arginine; Back; Biological; Biology; Cell Differentiation process; Cell Line; Cell Proliferation; Cells; Defect; Development; Exons; Eye; Genes; Genetic; Goals; Modeling; Mus; Mutant Strains Mice; Neural Crest; Neural Crest Cell; Numbers; Optics; Pathogenesis; Pigments; Post-Translational Protein Processing; Process; Protein Isoforms; Protein Splicing; Proteins; RNA Splicing; Regulation; Role; Serine; System; Tissues; Transcription factor genes; base; feeding; mutant; neuroepithelium; promoter; transcription factor

Alternative splicing, by which protein isoforms differing in sequence and in function can be generated from a single gene, is a mechanism important for the regulation of most if not all aspects of biology. Not surprisingly, splicing alterations are critical contributors to pathogenesis. We are currently studying alternative splicing in the transcription factor gene Mitf. This gene regulates the formation of pigment cells from the neural crest and the neuroepithelium of the eye and serves as an excellent model for assessing the biological significance of alternative splicing. We have generated a line of mice in which a change in an exonic sequence leads to a splicing alteration that eliminates the corresponding exon. Interestingly, the altered proteins affect cell proliferation differently compared to wild type. To dissect the underlying mechanisms, we have analyzed the cell proliferation parameters by FACS analysis and are studying the splicing defects in transfected minigenes. The studies indicate that the mutant minigenes recapitulate the splicing defects seen in the mutant mice and show that serine-arginine-rich splicing proteins, called SR proteins, enhance or suppress the splicing defect not only in the minigenes but also in the endogenous gene. Hence, by focusing on MITF and SR proteins, we have identified an interplay between splicing and cell proliferation, and we are currently focusing on the question of potential feed back loops that might exist between the two processes.

替代剪接，通过该剪接可以通过单个基因产生序列和功能不同的蛋白质同工型，这对于调节生物学的大多数（即使不是所有方面）都很重要。毫不奇怪，剪接改变是发病机理的关键因素。我们目前正在研究转录因子基因MITF中的替代剪接。该基因调节来自神经rest和眼睛神经上皮的色素细胞的形成，是评估替代剪接的生物学意义的绝佳模型。我们已经生成了一系列小鼠，其中外显子序列的变化会导致剪接变化，从而消除了相应的外显子。有趣的是，与野生型相比，改变的蛋白质影响细胞增殖。为了剖析潜在机制，我们通过FACS分析分析了细胞增殖参数，并正在研究转染的微型烯中的剪接缺陷。研究表明，突变体小组概括了突变小鼠中看到的剪接缺陷，并表明丝氨酸 - 精氨酸的剪接蛋白（称为SR蛋白）不仅在微型元素中，而且在内源基因中增强或抑制剪接缺陷。因此，通过关注MITF和SR蛋白，我们已经确定了剪接和细胞增殖之间的相互作用，并且目前我们正在关注两个过程之间可能存在的潜在馈回循环的问题。