Characterization of small open reading frames (sORFs) that encode for proteins 2

我们确定<I>S. 酿酒</I>。与Jef Boeke（The Johns）的实验室合作， 霍普金斯大学）、罗恩戴维斯（斯坦福大学）和迈克尔斯奈德（耶鲁大学），我们 构建了140个新鉴定的sORF的基因缺失突变体集合，并整合 将它们添加到现有的删除集合中。我们的实验室进行了全面分析 的sORF缺失菌株，并确定了22个sORF所需的单倍体生长，生长在 高温、在不可发酵的碳源存在下生长、或在可发酵的碳源中生长。 DNA损伤和复制抑制剂的存在。我们对sORF的有限分析 基于窄范围的实验条件和与当前序列的比较， 在数据库中可用。我们建议将额外的sORF确定为 数据库扩展。在继续我们正在进行的研究目标，我们发现， 新发现的sORF被确定为全球质量中的动粒组分 光谱研究。该结果表明这些sORF可能在以下方面发挥作用： 动粒功能，并表明额外的sORFs在 动粒功能和基因组稳定性。我们将分析sORF缺失菌株， 染色体丢失和检查点功能缺陷，并进行二次遗传筛查， 进一步确定sORF的分子作用。根据目前的结果，我们发现 鞘脂生物合成与 <I>MEC 1</I>介导的对DNA损伤的反应，并已确定了 通过细胞周期的G1-S期进展的新调节剂。在努力定义 sORF的其他作用，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。

We determined that there are 299 sORFs in<I>S. cerevisiae</I>. 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. Based on our results so far, we have uncovered a novel relationship between sphingolipid biosynthesis and <I>MEC1</I>-mediated response to DNA damage and have identified a new regulator for progression through the G1-S phase of the cell cycle. 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.