Impact of Non-canonical Decoding on the Proteome

描述（由申请人提供）：A。项目摘要/摘要翻译描述了mRNA翻译的局部重编程，以丢弃标准翻译规则并非规范地解码。在所有3个结构域中都观察到了这种刺激过程的产物，即在阅读框中掺入移位的蛋白质、绕过的核苷酸片段和/或意外的氨基酸，并且已知其对基因表达的转录后调节有重要贡献。我们的假设是，重新编码和错误编码的蛋白质比以前预期的更重要，但它们一直被高通量蛋白质组学方法所忽视，这些方法将蛋白质组视为基因组的一面镜子。我们相信，改变蛋白质组学工作流程可以揭示蛋白质在体内重新编码和错误编码，并且这些方法可以提供一些重新编码产物的氨基酸序列。如果成功，这些努力可能会影响蛋白质翻译，微生物学和蛋白质组学的研究。它们可以影响遗传性疾病治疗的发展，并可以解释某些基因产物的异常活性差异，这些基因产物仅因同义多态性而不同。它们也可能暗示了假信使RNA的功能。B。项目叙述本提案描述了一种方法，以检查合成生命基本成分的替代途径的重要性。如果发现该途径比以前预期的更重要，它将为某些疾病提供病因和潜在的治疗方法。

DESCRIPTION (provided by applicant): A. Project Summary/Abstract Recoding describes the local reprogramming of mRNA translation to discard standard translational rules and decode non-canonically. The products of this stimulated process, proteins incorporating shifts in reading frame, bypassed nucleotide segments, and/or unexpected amino acids have been observed in all 3 domains and are known to contribute importantly to post-transcriptional regulation of gene expression. Our hypothesis is that recoded and miscoded proteins are more important than previously anticipated, but that they are consistently overlooked by high-throughput proteomic methods that regard the proteome as a mirror of the genome. We believe that an altered proteomic workflow can reveal proteins recoded and miscoded in vivo, and that these methods can provide amino acid sequences for some of the recoded products. If successful, these efforts can potentially impact research in protein translation, microbiology, and proteomics. They can impact the development of therapies for genetic diseases, and can suggest explanations for the unusual activity differences observed for some gene products differing only by a synonymous polymorphism. They may also suggest a function for pseudo-messenger RNA. B. Project Narrative This proposal describes a method to examine the importance of an alternate pathway for synthesizing essential components of life. Should the pathway be found to be more important than previously anticipated, it will suggest causes and potential therapies for some disorders.