Characterization of Small Open Reading Frames (sORFs) that Encode for Proteins

利用来自独立实验方法和计算分析的数据， 提出了IS中存在299个sORF。 cerevisiae/I基因组，占注释ORF的约5%。我们 确定在其他真核生物中注释了类似百分比的sORF，包括 人类和184个I S酿酒酵母/I sORFs 与其他生物的ORF相似。与杰夫的实验室合作， Boeke（约翰·霍普金斯大学）、罗恩·戴维斯（斯坦福大学）和迈克尔·斯奈德 (Yale我们构建了140个新的基因缺失突变体的集合， 鉴定了sORF并将其整合到现有的删除集合中。本实验室 对sORF缺失菌株进行了综合分析，鉴定出22个sORF 所需的单倍体生长，生长在高温下，生长在存在的一个 不可发酵的碳源，或在DNA损伤和复制停滞的存在下生长 剂.我们对sORF的有限分析是基于小范围的实验结果。 条件和与数据库中当前可用序列的比较。我们建议 随着数据库的扩展，将鉴定出另外的sORF。为了继续我们的 研究目的我们发现，三个新发现的sORF已被确定为 全球质谱研究中的动粒组分。这一结果表明，这些 sORFs可能在动粒功能中起作用，并提示 在动粒功能和基因组稳定性中的额外sORF。我们将分析sORF 缺失菌株的染色体丢失和检查点功能的缺陷，并进行二次 基因筛选以进一步确定sORF的分子作用。在努力定义其他 sORF的作用，我们有几个正在进行的合作，以检查是否sORF删除 菌株具有与细胞周期进程、核转运、基因表达和细胞周期缺陷相关的表型。 沉默和转录调节。我们还产生了表位标记的sORF菌株， 以及过量表达sORF的质粒供公众使用。我们对sORF的研究将识别和 确定sORF在不同生物途径中的作用。我们建议，额外的sORF 将随着数据库的扩展而被识别。我们将利用计算，生物化学， 基因组方法来验证sORF的存在并了解其生物学功能。

Utilizing data from independent experimental approaches and computational analyses we proposed the existence for 299 sORFs in the <I>S. cerevisiae</I> genome, representing about 5% of annotated ORFs. We determined that a similar percentage of sORFs are annotated in other eukaryotes, including humans, and 184 of the <I>S. cerevisiae</I> sORFs exhibit similarity with ORFs in other organisms. In collaboration with the laboratories of Jef Boeke (The Johns Hopkins University), Ron Davis (Stanford University), and Michael Snyder (Yale University), we constructed a collection of gene-deletion mutants of 140 newly identified sORFs and integrated them into the existing deletion collection. Our laboratory undertook a comprehensive analysis of the sORF deletion strains and identified 22 sORFs required for haploid growth, growth at high temperature, growth in the presence of a non-fermentable carbon source, or growth in the presence of DNA damage and replication arrest agents. Our limited analysis of the sORFs was based on a narrow range of experimental conditions and comparison to sequences currently available in the databases. We propose that additional sORFs will be identified as the databases expand. In continuation with our ongoing research objective we found that three of the newly identified sORFs have been identified as kinetochore components in global mass spectrometric studies. This result indicates that these sORFs likely have a role in kinetochore function and suggests the potential for the role of additional sORFs in kinetochore function and genome stability. We will analyze the sORF deletion strains for chromosome loss and defects in checkpoint function, and perform secondary genetic screens to further define the molecular role of the sORFs. In efforts to define other roles for sORFs, we have several ongoing collaborations to examine if the sORF deletion strains have phenotypes related to defects in cell cycle progression, nuclear transport, gene silencing and transcriptional regulation. We have also generated epitope tagged sORF strains and plasmids overexpressing the sORFs for public use. Our studies with sORFs will identify and establish the role of sORFs in diverse biological pathways. We propose that additional sORFs will be identified as the databases expand.. We will utilize computational, biochemical, and genomic approaches to validate the presence of sORFs and understand their biological function.