PROJECT 4

项目4：体内蛋白质翻译后修饰的光学监测（Thorn）（#27） 我们开发了利用荧光共振能量研究蛋白质修饰和相互作用的工具 转移（FRET）显微术。主要的努力是检测wVo 12中的蛋白质-蛋白质相互作用，并使用这些 数据来预测大型蛋白质复合物的结构。我们开发了一种方法， 从酵母的可重复的FRET数据，并使用它们来获得septin复合物的低分辨率结构 （这是从酵母到哺乳动物的细胞分裂所必需的）。在Michael Laub（项目7）中， 在两种成分的信号蛋白之间，翻译后调节的主要参与者， 细菌，并与Daughterly和他的同事合作，开发高通量，定量，FRET为基础的 鉴定蛋白质相互作用的方法13.库尔特·索恩，PI，曾是鲍尔研究员，现任 加州大学旧金山分校尼康成像中心 新项目4：基因中的串联重复序列：适应的高变模块？（Verstrepen）（#28-30） 我们验证了一个假设，即在芽殖酵母中，串联重复序列是高度可变的遗传基因。 模块，其可变性有助于生物体适应不断变化的环境。大多数测序的基因组 含有串联重复序列;其中一些出现在编码序列中，重组可以改变编码序列的数量。 在翻译蛋白质中的肽重复序列。我们开发了一种算法， (http：//hulsweb1.cgr.harvard.edu/SERV/）来查找和注释重复，预测它们在数量上是否变化， 将它们与其他信息（表达水平，与转录因子结合位点的重叠， 核小体位点等），并将这些数据存款一个可公开访问的数据库 (http：//hulsweb1.cgr.harvard.edu/TandemRepeat/），可以快速自动地与现有的 数据库; 2）产生在启动子中具有不同数目的重复和开放阅读的酵母菌株 选定的基因框架，并研究了这种变化的表型后果（正在进行）; 3）研究 在拟南芥和水稻中（与C. Queitsch，new project 7）.我们用了两个 对基因组进行测序，以找到所有含有可变重复数量的ORF，选择150个基因， 描述了它们在12个A.从不同的地方分离到了70多株， 在不同分离株之间显示重复数变化的基因。凯文·弗斯特莱彭成为鲍尔家族的一员 2005年成为研究员，2006年加入CMB。

Project 4: Optical monitoring of post-translational modification of proteins in vivo (Thorn)(#27) We developed tools to study protein modification and interactions using fluorescence resonance energy transfer (FRET) microscopy. The main effort was to detect protein-protein interactions in wVo12 and use these data to predict the structure of large protein complexes. We developed methods to produce precise and reproducible FRET data from yeast, and used them to derive a low-resolution structure for the septin complex (which is required for cell division from yeast to mammals). With Michael Laub (project 7) we found interactions between two-component signaling proteins, the predominant participants in post-translational regulation in bacteria, and collaborated with Daugherty and his colleagues to develop high throughput, quantitative, FRETbased methods to identify protein interactions13. Kurt Thorn, the PI, was a Bauer Fellow and is now Director of the Nikon Imaging Center at UCSF. New project 4: Tandem repeats in genes: hyper-variable modules for adaptation? (Verstrepen) (#28-30) We tested the hypothesis35 that in budding yeast, tandemly repeated sequences are hyper-variable genetic modules, whose variability helps organisms adapt to changing environments. Most sequenced genomes contain tandem repeats; some of these occur in coding sequences, where recombination can alter the number of peptide repeats in translated proteins. We have, 1) developed an algorithm36 (http://hulsweb1.cgr.harvard.edu/SERV/) to find and annotate repeats, predict whether they vary in number, associate them with other information (expression level, overlap with transcription factor binding sites, nucleosome sites etc.), and deposit this data in a publicly accessible database (http://hulsweb1.cgr.harvard.edu/TandemRepeat/) that allows rapid and automated comparison to existing databases; 2) generated yeast strains with different numbers of repeats in the promoters and open reading frames of selected genes, and studied the phenotypic consequences of this variation (ongoing); 3) studied tandem repeats in Arabidopsis thaliana and Oryza sativa (with C. Queitsch, new project 7). We have used two sequenced genomes to find all ORFs containing variable numbers of repeats, selected 150 genes, characterized their variation across twelve A. thaliana isolates from different locations, and found more than 70 genes that show variation in repeat number between different isolates. Kevin Verstrepen became a Bauer Fellow in 2005 and joined the CMB in 2006.