Systematic Discovery and Analysis of Small Proteins and Small ORFs in Mycobacteria

分枝杆菌中小蛋白和小 ORF 的系统发现和分析

• 批准号: 10388045
• 负责人: KEITH M DERBYSHIRE
• 金额: $ 0.89万
• 依托单位: WADSWORTH CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10388045
• 关键词: Algorithms; Amino Acids; Basic Science; Biochemical; Birth; Cell Extracts; Cysteine; Eukaryota; Foxes; Friends; Future; Genes; Genome; Genomics; Genus Mycobacterium; Goals; Homeostasis; Laboratory Research; Learning; Mass Spectrum Analysis; Mediating; Mentors; Molecular Biology; Mutation; Open Reading Frames; Peptides; Preparation; Prokaryotic Cells; Property; Proteins; Proteomics; Regulation; Research; Science; Scientist; Training; Work; career; experience; genome annotation; mutant; skills; summer research; undergraduate research experience; undergraduate student; whole genome

The last few decades have seen the birth and maturation of the field of Molecular Biology. Initially, mutant genes were focal points of genome exploration. Now, entire genomes are routinely sequenced, and the resident genes are automatically identified by annotation algorithms. Alternatively, proteomic approaches prepare proteolytic peptides of whole-cell extracts for analysis by mass spectrometry. Each of these approaches are strongly biased for large genes: large genes are frequent targets for mutation, long-open- reading frames are easily discerned in genomic sequence, and large proteins generate many peptides for mass spectrometry identification. This unintended bias has also created a large gap in our understanding of molecular biology. Recent work in eukaryotes and prokaryotes alike have uncovered multitudes of small genes or their encoded proteins. The numbers of small proteins (considered as 50 aa or less) rival that of traditionally large proteins, yet only a handful have been ascribed a function. The goal of this proposal is to propel this nascent field forward by facilitating both small protein discovery and functional characterization. We will develop and apply our small protein approaches in mycobacteria. Mycobacteria offer many advantages for small protein study. Foremost is that they express >1000 small proteins in standard conditions. The goal of this supplement is to support an outstanding undergraduate student, Lauren Fox, who aspires to be a research scientist, with direct hands-on laboratory research experience. Lauren will focus on dissecting the mechanism of regulation mediated by a unique set of sproteins involved in cysteine homeostasis. She will learn new technical and analytical skills, and develop a greater understanding of what it takes to be a research scientist. Involvement in the Wadsworth summer research program for undergraduates will further expose Lauren to potential careers in science, future mentors and new friends and colleagues.

在过去的几十年中，分子生物学领域的出生和成熟。最初，突变体 基因是基因组探索的焦点。现在，整个基因组常规测序，并且 居民基因通过注释算法自动鉴定。或者，蛋白质组学方法 制备全细胞提取物的蛋白水解肽，以通过质谱法分析。每个 对于大基因而言，方法有很大的偏见：大基因是突变的频繁靶标，长期开放 读取帧很容易以基因组序列辨别，并且大蛋白会产生许多肽 用于质谱识别。这种意外的偏见也在我们的 了解分子生物学。 真核生物和原核生物的最新工作都发现了许多小基因或其它们 编码的蛋白质。传统上，小蛋白质的数量（被认为是50 aa或以下）与 大型蛋白质，但只有少数属性归因于功能。该提议的目的是推动 通过促进小蛋白质发现和功能表征，这一新生场的前进。我们 将在分枝杆菌中发展并应用我们的小蛋白质方法。分枝杆菌提供了许多 小蛋白质研究的优势。最重要的是它们在标准中表达> 1000个小蛋白 状况。 该补充的目的是支持一位杰出的本科生劳伦·福克斯（Lauren Fox） 成为一名具有直接动手实验室研究经验的研究科学家。劳伦将专注于 解剖由半胱氨酸参与的独特的链霉素介导的调节机制 稳态。她将学习新的技术和分析技能，并对 成为研究科学家需要什么。参与Wadsworth夏季研究计划 本科生将进一步使劳伦（Lauren）接触科学，未来的导师和新朋友的潜在职业 和同事。