Arabadopsis 2010: Identification of New Plastid Division Genes in Arabidopsis and Comprehensive Analysis of Their Functions

植物的质体分裂是由一个复杂的大分子装置来协调的，其组成成分大多是未知的。将确定一组预测的和遗传上确定的可能参与质体分裂的拟南芥基因产物的功能。待研究的蛋白质来源于三个来源：1）与已知的质体分裂、细菌细胞分裂、线粒体分裂或融合、或基于微管的马达蛋白具有序列或结构同源性的预测的蛋白质; 2）由拟南芥弧定义的基因，（叶绿体的积累和复制）突变体;和3）编码与蓝细菌细胞分裂蛋白同源的蛋白质的基因，将通过聚球藻PCC 7942中的转座子诱变来鉴定。功能研究将包括通过显微镜检查叶绿体数量，大小，形状，超微结构和FtsZ定位模式的异常，蛋白质靶向和拓扑学的生化测定，使用GFP融合蛋白和蛋白质特异性抗体分析体内定位，以及使用免疫沉淀和酵母双杂交技术分析蛋白质相互作用。从功能调查获得的结果将被整合到一个模型，描述的大分子组织和生化活性的叶绿体分裂复合体，新的假设可以推导和测试。这项研究将与对各种生物学问题感兴趣的科学家有关，包括细胞器裂变和动力学，大分子组织，细胞器进化和植物发育。在这些研究中产生的cDNA克隆、质粒、抗体和其他有用的试剂将提供给更广泛的科学界。功能信息将通过公布结果和通过网站（http：//www.msu.edu/cnosteryou/2010.htm）进行交流，网站将定期更新，以反映获得的敲除和arc突变表型数据。随着获得新信息，将修订数据库注释。除了为研究生和博士后学者提供培训机会外，该项目预计还将为一些本科生，包括代表性不足群体的成员提供研究经验。

Plastid division in plants is orchestrated by a complex macromolecular apparatus, the components of which are mostly unknown. The functions of a set of predicted and genetically defined Arabidopsis gene products likely to participate in plastid division will be determined. The proteins to be investigated derive from three sources: 1) predicted proteins having sequence or structural homology to known plastid division, bacterial cell division, mitochondrial fission or fusion, or microtubule-based motor proteins; 2) genes defined by the Arabidopsis arc (accumulation and replication of chloroplasts) mutants; and 3) genes encoding proteins with homology to cyanobacterial cell division proteins, to be identified by transposon mutagenesis in Synechococcus PCC 7942. Functional studies will include examination of knockout lines by microscopy for abnormalities in chloroplast number, size, shape, ultrastructure and FtsZ localization pattern, biochemical assays of protein targeting and topology, analysis of in vivo localization using GFP fusion proteins and protein-specific antibodies, and analysis of protein interactions using immunoprecipitation and yeast two-hybrid techniques. The results obtained from functional investigations will be integrated to arrive at a model describing the macromolecular organization and biochemical activity of the chloroplast division complex from which new hypotheses can be derived and tested. This research will be relevant to scientists interested in a variety of biological problems, including organelle fission and dynamics, macromolecular organization, organelle evolution, and plant development. cDNA clones, plasmids, antibodies and other useful reagents generated during these studies will be made available to the broader scientific community. Functional information will be communicated through publication of results and through a website (http://www.msu.edu/~osteryou/2010.htm) to be updated regularly to reflect data obtained on knockout and arc mutant phenotypes. Database annotations will be revised as new information is obtained. In addition to providing training opportunities to graduate students and postdoctoral scholars, the project is expected to provide research experience to several undergraduate students, including members of underrepresented groups.