Impact of Non-canonical Decoding on the Proteome

项目概述/摘要重编码描述mRNA翻译的局部重编程，抛弃标准翻译规则，进行非规范解码。这一刺激过程的产物，包含阅读框移位的蛋白质，绕过的核苷酸片段，和/或意想不到的氨基酸，已经在所有3个结构域中观察到，并且已知对基因表达的转录后调控有重要贡献。我们的假设是，重新编码和错误编码的蛋白质比先前预期的更重要，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镜子。我们相信，改变蛋白质组学工作流程可以揭示体内重新编码和错误编码的蛋白质，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。它们可以影响遗传疾病治疗的发展，并可以解释一些基因产物的不寻常的活性差异，这些差异仅通过同义多态性来观察。它们也可能暗示了伪信使RNA的功能。本提案描述了一种方法来检验合成生命基本成分的另一种途径的重要性。如果发现该通路比先前预期的更重要，它将提示一些疾病的原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.