Abstract
The optically active form of tritium-labeled A-factor (2-isocapryloyl-3R-hydroxymethyl-gamma-butyrolactone), a pleiotropic autoregulator responsible for streptomycin production, streptomycin resistance, and sporulation in Streptomyces griseus, was chemically synthesized. By using the radioactive A-factor, a binding protein for A-factor was detected in the cytoplasmic fraction of this organism. The binding protein had an apparent molecular weight of approximately 26,000, as determined by gel filtration. Scatchard analysis suggested that A-factor bound the protein in the molar ratio of 1:1 with a binding constant, Kd, of 0.7 nM. The number of the binding protein was roughly estimated to be 37 per genome. The "inducing material" virginiae butanolide C (VB-C), which has a structure very similar to that of A-factor and is essential for virginiamycin production in Streptomyces virginiae, did not inhibit binding. In addition, no protein capable of specifically binding 3H-labeled VB-C was found in S. griseus. Together with the observation that VB-C had almost no biological activity on the restoration of streptomycin production or sporulation in an A-factor-deficient mutant of S. griseus, these results indicated that the binding protein had a strict ligand specificity. Examination for an A-factor-binding protein in Streptomyces coelicolor A3(2) and Streptomyces lividans showed the absence of any specifically binding protein.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Distler J., Ebert A., Mansouri K., Pissowotzki K., Stockmann M., Piepersberg W. Gene cluster for streptomycin biosynthesis in Streptomyces griseus: nucleotide sequence of three genes and analysis of transcriptional activity. Nucleic Acids Res. 1987 Oct 12;15(19):8041–8056. doi: 10.1093/nar/15.19.8041. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gräfe U., Schade W., Eritt I., Fleck W. F., Radics L. A new inducer of anthracycline biosynthesis from Streptomyces viridochromogenes. J Antibiot (Tokyo) 1982 Dec;35(12):1722–1723. doi: 10.7164/antibiotics.35.1722. [DOI] [PubMed] [Google Scholar]
- Hara O., Beppu T. Induction of streptomycin-inactivating enzyme by A-factor in Streptomyces griseus. J Antibiot (Tokyo) 1982 Sep;35(9):1208–1215. doi: 10.7164/antibiotics.35.1208. [DOI] [PubMed] [Google Scholar]
- Hara O., Beppu T. Mutants blocked in streptomycin production in Streptomyces griseus - the role of A-factor. J Antibiot (Tokyo) 1982 Mar;35(3):349–358. doi: 10.7164/antibiotics.35.349. [DOI] [PubMed] [Google Scholar]
- Hara O., Horinouchi S., Uozumi T., Beppu T. Genetic analysis of A-factor synthesis in Streptomyces coelicolor A3(2) and Streptomyces griseus. J Gen Microbiol. 1983 Sep;129(9):2939–2944. doi: 10.1099/00221287-129-9-2939. [DOI] [PubMed] [Google Scholar]
- Horinouchi S., Hara O., Beppu T. Cloning of a pleiotropic gene that positively controls biosynthesis of A-factor, actinorhodin, and prodigiosin in Streptomyces coelicolor A3(2) and Streptomyces lividans. J Bacteriol. 1983 Sep;155(3):1238–1248. doi: 10.1128/jb.155.3.1238-1248.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Horinouchi S., Kumada Y., Beppu T. Unstable genetic determinant of A-factor biosynthesis in streptomycin-producing organisms: cloning and characterization. J Bacteriol. 1984 May;158(2):481–487. doi: 10.1128/jb.158.2.481-487.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Horinouchi S., Suzuki H., Nishiyama M., Beppu T. Nucleotide sequence and transcriptional analysis of the Streptomyces griseus gene (afsA) responsible for A-factor biosynthesis. J Bacteriol. 1989 Feb;171(2):1206–1210. doi: 10.1128/jb.171.2.1206-1210.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Khokhlov A. S., Anisova L. N., Tovarova I. I., Kleiner E. M., Kovalenko I. V., Krasilnikova O. I., Kornitskaya E. Y., Pliner S. A. Effect of A-factor on the growth of asporogenous mutants of Streptomyces griseus, not producing this factor. Z Allg Mikrobiol. 1973;13(8):647–655. doi: 10.1002/jobm.3630130803. [DOI] [PubMed] [Google Scholar]
- Khokhlov A. S., Tovarova I. I., Borisova L. N., Pliner S. A., Shevchenko L. N., Kornitskaia E. Ia, Ivkina N. S., Rapoport I. A. A-faktor, obespechivaiushchii biosintez streptomitsina mutantnym shtammom Actinomyces streptomycini. Dokl Akad Nauk SSSR. 1967 Nov-Dec;177(1):232–235. [PubMed] [Google Scholar]
- Nihira T., Shimizu Y., Kim H. S., Yamada Y. Structure-activity relationships of virginiae butanolide C, an inducer of virginiamycin production in Streptomyces virginiae. J Antibiot (Tokyo) 1988 Dec;41(12):1828–1837. doi: 10.7164/antibiotics.41.1828. [DOI] [PubMed] [Google Scholar]
- SAITO H., MIURA K. I. PREPARATION OF TRANSFORMING DEOXYRIBONUCLEIC ACID BY PHENOL TREATMENT. Biochim Biophys Acta. 1963 Aug 20;72:619–629. [PubMed] [Google Scholar]
- Yamada Y., Sugamura K., Kondo K., Yanagimoto M., Okada H. The structure of inducing factors for virginiamycin production in Streptomyces virginiae. J Antibiot (Tokyo) 1987 Apr;40(4):496–504. doi: 10.7164/antibiotics.40.496. [DOI] [PubMed] [Google Scholar]