| 1.Research Institution | University of Tokyo | |
| 2.Research Area | Integrated Fields | |
| 3.Research Field | Production of Novel Useful Substances through Integration of Life Sciences and Chemistry | |
| 4.Term of Project | FY1996〜FY2000 | |
| 5.Project Number | 96I00303 | |
| 6.Title of Project | Molecular analysis of the initiation of secondary metabolite formation and its application to molecular breeding of producers |
| Name | Institution,Department | Title of Position |
| Horinouchi, Sueharu | Univ. of Tokyo, Graduate School of Agricultural & Life Sciences | Professor |
8.Core Members
| Names | Institution,Department | Title of Position |
| Yamada, Yasuhiro | Osaka Univ., Graduate School of Engineering | Professor |
| Nihira, Takuya | Osaka Univ., Graduate School of Engineering | Assoc. Professor |
| Beppu, Teruhiko | Nihon Univ., College of Bioresource Sciences | Professor |
9.Cooperating Researchers
| Names | Institution,Department | Title of Position |
| Ohnishi, Yasuo | Univ. of Tokyo, Graduate School of Agricultural & Life Sciences | Assistant |
| Ueda, Kenji | Nihon Univ., College of Bioresource Sciences | Assistant |
| Ihara, Fumio | Osaka Univ., Graduate School of Engineering | Assistant |
10.Summary of Research Results
|
(1) The A-factor-specific receptor served as a repressor-type regulator for adpA encoding a transcriptional activator of many genes required for secondary metabolism and morphological differentiation. The regulatory cascade leading to streptomycin biosynthesis has been elucidated. An A-factor-dependent sigma factor of RNA polymerase essential for morphogenesis was also identified. Additional regulatory systems including eukaryotic-type protein phosphorylation, cAMP, and calmodulin were found to control secondary metabolism and morphogenesis and cross-talk with the A-factor regulatory cascade. (2) Virginiae butanolide-specific receptor protein (BarA) and IM-2 specific receptor protein (FarA) were purified from Streptomyces virginiae and Streptomyces lavendulae FRI-5, respectively. With the corresponding genes, recombinant receptors were overexpressed in E. coli, and in vitro analyses using the purified receptor revealed the high affinity (Kd= 〜lnM) and the high ligand specificity of the receptors. Phenotypic analyses of null mutants of the receptor genes as well as the functional analyses of genes surrounding the VB receptor gene (barA) clarified the detailed regulation mechanism on the VB-mediated expression of antibiotic resistance together with the overall signalling cascade leading to the antibiotic biosynthesis. (3) As environmental factors affecting the initiation of secondary metabolism and morphological differentiation in Streptomyces, we characterized not only the involvement of carbon-sources and copper, but also the interspecific stimulation of those events mediated by putative diffusible signal substances, which is widely distributed among Streptomyces. As for the former subject, we cloned related genes and generated knockdown mutants, which revealed the role of those genes. For the latter, we purified several chemical substances that possibly work as the interspecific signals. Furthermore, we identified and inactivated other novel regulatory genes for secondary metabolism or morphogenesis. All results enabled us to establish a fundamental standpoint for studying the effect by various environmental factors on secondary metabolism and cellular differentiation in Streptomyces. |
11.Key Words
(1)Streptomyces、(2)autoregulatory factors、(3)antibiotic production
(4)morphogenesis、(5)control of gene expression、(6)microbial hormone
(7)A-factor
12.References
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| Journal | Volume | Year | Pages Concerned | |
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| Microbiology | 145 | 1999 | 2265-2272 | |
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| Journal | Volume | Year | Pages Concerned | |
| Actinomycetologica | 13 | 1999 | 89-93 | |
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| Ueda, K. | Carbon-source-dependent transcriptional control involved in the initiation of cellular differentiation in Streptomyces griseus. | |||
| Journal | Volume | Year | Pages Concerned | |
| Antonie van Leeuwenhoek | 2000 | in press | ||
| Author | Title of Book | ||
| Horinouchi, S. | Dynamic Aspects of Natural Products Chemistry: Molecular Biological Approaches | ||
| Publisher | Year | Pages | |
| Kodansha and Harwood Academic Publishers | 1997 | 165-186 | |
| Author | Title of Book | ||
| Horinouchi, S. | Proceedings of the International Symposium on Bioactive Compounds and Strain Development | ||
| Publisher | Year | Pages | |
| The Korean Society for Applied Microbiology | 1998 | 66-72 | |
| Author | Title of Book | ||
| Horinouchi, S. | Cell-Cell Signaling in Bacteria | ||
| Publisher | Year | Pages | |
| American Society for Microbiology | 1999 | 93-207 | |
| Author | Title of Book | ||
| Yamada, Y. | Biotechnology of Antibiotics | ||
| Publisher | Year | Pages | |
| Marcel Dekker, Inc | 1999 | 63-79 | |
| Author | Title of Book | ||
| Yamada, Y. | Comprehensive Natural Products Chemistry | ||
| Publisher | Year | Pages | |
| Elsevier Science | 1998 | 377-413 | |