KANATA Shigehiko Nara Institute of Science and Technology, Graduate School of Information Science, Professor, 情報科学研究科, 教授 (90224584)
KAWAMURA Fujio Rikkyo University, Faculty of Science, Professor, 理学部, 教授 (10126039)
YOSHIKAWA Hirofumi Tokyo University of Agriculture, Department of Bioscience, Professor, 応用生物科学部, 教授 (50175676)
WATABE Kazuhito Setsunan Univaersity, Faculty of Pharmaceutical Sciences, Professor, 薬学部, 教授 (20107105)
SATO Thutomu Tokyo University of Agriculture and Technology, Graduate School of Agriculture, Associate Professor, 共生科学技術研究部, 助教授 (70215812)
1) We newly constructed about 300 mutants of the genes identified by genome sequencing, to complete the international mutant construction project, and reported that 271 genes are essential for the B. subtilis growth at 37℃ in LB medium.
2) Among essential genes without known function, we have found that yacA encodes an RNA-modifying enzyme responsible for lysidine formation and yneS, together with plsX, is essential for the first step of phospholipids biosynthesis. In addition, one of the essential GTP binding proteins, YlqF, has been demonstrated to participate in the last step of 50S ribosome subunit assembly.
3) We have attempted to construct knock out mutant of each of 57 ribosome protein genes, and found that 22 are dispensable for the growth.
4) We have systematically analyzed the synthetic lethality of double knockout of paralogous gene pairs, and found that the polA-yacP double mutant is lethal due to the defect in removal of primer RNA of Okazaki fragments.
5) By combination of proteome, transcriptome and genetic analysis, in addition to 119 genes previously reported, we have newly identified 175 genes that are expressed specifically during sporulation, and determined sigma factors responsible for their expression. We have also analyzed the localization of GFP fusions of about 90 spore proteins, and found at least 10 different the localization patterns.
6) We have attempted the analysis of protein-protein interaction network by using yeast two-hybrid analysis and mass spectroscopic analysis of in vivo protein complex, and identified several interesting interactions; anti-sigma proteins regulating activities of ECF sigma factors, Rap proteins regulating ComK responsible for competence development, YheIH involved in regulation of phosphor-relay inducing initiation of sporulation, YlqF interacting with FtsZ and involved in cell division, and YabA interacting with DnaA and DnaA and regulating initiation of genome replication.