Abstract
Ribulose bisphosphate carboxylase is essential for both photoautotrophic and photoheterotrophic growth of the cyanobacterium Synechocystis 6803. However, a mutant lacking cyanobacterial carboxylase could be obtained by replacing the natural carboxylase gene with the corresponding gene from Rhodospirillum rubrum, a photosynthetic anaerobe. This treatment produced an organism whose growth depended on the activity of the structurally and functionally dissimilar foreign carboxylase. As a further consequence of this mutagenic replacement, the mutant also lacked microscopically observable carboxysomes, the subcellular inclusion bodies in which the wild-type carboxylase naturally resides. The mutant, dependent on a carboxylase with an inferior relative specificity for CO2 versus O2 and apparently lacking carboxysomes, is extremely sensitive to the CO2/O2 ratio supplied during growth and is unable to grow at all in air. This response to the gas composition should prove useful for selection of various R. rubrum carboxylase mutants with altered specificities for CO2 and O2.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Chauvat F., Rouet P., Bottin H., Boussac A. Mutagenesis by random cloning of an Escherichia coli kanamycin resistance gene into the genome of the cyanobacterium Synechocystis PCC 6803: selection of mutants defective in photosynthesis. Mol Gen Genet. 1989 Mar;216(1):51–59. doi: 10.1007/BF00332230. [DOI] [PubMed] [Google Scholar]
- Curtis S. E., Haselkorn R. Isolation and sequence of the gene for the large subunit of ribulose-1,5-bisphosphate carboxylase from the cyanobacterium Anabaena 7120. Proc Natl Acad Sci U S A. 1983 Apr;80(7):1835–1839. doi: 10.1073/pnas.80.7.1835. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lambert D. H., Stevens S. E., Jr Photoheterotrophic growth of Agmenellum quadruplicatum PR-6. J Bacteriol. 1986 Feb;165(2):654–656. doi: 10.1128/jb.165.2.654-656.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Oka A., Sugisaki H., Takanami M. Nucleotide sequence of the kanamycin resistance transposon Tn903. J Mol Biol. 1981 Apr 5;147(2):217–226. doi: 10.1016/0022-2836(81)90438-1. [DOI] [PubMed] [Google Scholar]
- Pelroy R. A., Rippka R., Stanier R. Y. Metabolism of glucose by unicellular blue-green algae. Arch Mikrobiol. 1972;87(4):303–322. doi: 10.1007/BF00409131. [DOI] [PubMed] [Google Scholar]
- Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
- Tabita F. R., McFadden B. A. D-ribulose 1,5-diphosphate carboxylase from Rhodospirillum rubrum. II. Quaternary structure, composition, catalytic, and immunological properties. J Biol Chem. 1974 Jun 10;249(11):3459–3464. [PubMed] [Google Scholar]
- Takabe T., Nishimura M., Akazawa T. Presence of two subunit types in ribulose-1,5-bisphosphate carboxylase from blue-green algae. Biochem Biophys Res Commun. 1976 Jan 26;68(2):537–544. doi: 10.1016/0006-291x(76)91179-7. [DOI] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vermaas W. F., Williams J. G., Rutherford A. W., Mathis P., Arntzen C. J. Genetically engineered mutant of the cyanobacterium Synechocystis 6803 lacks the photosystem II chlorophyll-binding protein CP-47. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9474–9477. doi: 10.1073/pnas.83.24.9474. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]