Abstract
The ggtA gene was sequenced during the analysis of a mutant of Synechocystis sp. strain PCC 6803 with impaired salt tolerance. It showed striking sequence similarities to ATP-binding proteins of binding-protein-dependent transport systems (ABC transporters). Mutants of ggtA and three neighboring reading frames were constructed by inserting an aphII gene cassette and were physiologically and genetically characterized. The ggtA insertion mutant lost its glucosylglycerol (GG) uptake ability, but its salt tolerance did not change. Therefore, it was concluded that active transport of the osmoprotective compound GG in Synechocystis is mediated by an ABC transporter. The genes for the GG-specific ABC transporter are not organized in an operon as usually found for comparable transporters, since the other insertion mutants showed normal GG transport activity. After cultivation of the ggtA mutant at high salt concentrations, significant amounts of GG were found in the cultivation medium, indicating that GG transport is mainly necessary for recovery of GG leaked through the cytoplasmic membrane. The Northern blot technique revealed increased transcription of the ggtA gene in cells adapted to higher salt concentrations, whereas in cells from basal medium, its transcription was weak.
Full Text
The Full Text of this article is available as a PDF (2.4 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ames G. F. Bacterial periplasmic transport systems: structure, mechanism, and evolution. Annu Rev Biochem. 1986;55:397–425. doi: 10.1146/annurev.bi.55.070186.002145. [DOI] [PubMed] [Google Scholar]
- Apte S. K., Haselkorn R. Cloning of salinity stress-induced genes from the salt-tolerant nitrogen-fixing cyanobacterium Anabaena torulosa. Plant Mol Biol. 1990 Nov;15(5):723–733. doi: 10.1007/BF00016122. [DOI] [PubMed] [Google Scholar]
- Bartsevich V. V., Pakrasi H. B. Molecular identification of an ABC transporter complex for manganese: analysis of a cyanobacterial mutant strain impaired in the photosynthetic oxygen evolution process. EMBO J. 1995 May 1;14(9):1845–1853. doi: 10.1002/j.1460-2075.1995.tb07176.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brown A. D. Microbial water stress. Bacteriol Rev. 1976 Dec;40(4):803–846. doi: 10.1128/br.40.4.803-846.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Csonka L. N., Hanson A. D. Prokaryotic osmoregulation: genetics and physiology. Annu Rev Microbiol. 1991;45:569–606. doi: 10.1146/annurev.mi.45.100191.003033. [DOI] [PubMed] [Google Scholar]
- Dayhoff M. O., Barker W. C., Hunt L. T. Establishing homologies in protein sequences. Methods Enzymol. 1983;91:524–545. doi: 10.1016/s0076-6879(83)91049-2. [DOI] [PubMed] [Google Scholar]
- Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fry D. C., Kuby S. A., Mildvan A. S. ATP-binding site of adenylate kinase: mechanistic implications of its homology with ras-encoded p21, F1-ATPase, and other nucleotide-binding proteins. Proc Natl Acad Sci U S A. 1986 Feb;83(4):907–911. doi: 10.1073/pnas.83.4.907. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Giaever H. M., Styrvold O. B., Kaasen I., Strøm A. R. Biochemical and genetic characterization of osmoregulatory trehalose synthesis in Escherichia coli. J Bacteriol. 1988 Jun;170(6):2841–2849. doi: 10.1128/jb.170.6.2841-2849.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gilson E., Nikaido H., Hofnung M. Sequence of the malK gene in E.coli K12. Nucleic Acids Res. 1982 Nov 25;10(22):7449–7458. doi: 10.1093/nar/10.22.7449. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gowrishankar J. Nucleotide sequence of the osmoregulatory proU operon of Escherichia coli. J Bacteriol. 1989 Apr;171(4):1923–1931. doi: 10.1128/jb.171.4.1923-1931.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Green L. S., Laudenbach D. E., Grossman A. R. A region of a cyanobacterial genome required for sulfate transport. Proc Natl Acad Sci U S A. 1989 Mar;86(6):1949–1953. doi: 10.1073/pnas.86.6.1949. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hagemann M., Richter S., Zuther E., Schoor A. Characterization of a glucosylglycerol-phosphate-accumulating, salt-sensitive mutant of the cyanobacterium Synechocystis sp. strain PCC 6803. Arch Microbiol. 1996 Aug;166(2):83–91. doi: 10.1007/s002030050360. [DOI] [PubMed] [Google Scholar]
- Kempf B., Bremer E. OpuA, an osmotically regulated binding protein-dependent transport system for the osmoprotectant glycine betaine in Bacillus subtilis. J Biol Chem. 1995 Jul 14;270(28):16701–16713. doi: 10.1074/jbc.270.28.16701. [DOI] [PubMed] [Google Scholar]
- Koonin E. V. A common set of conserved motifs in a vast variety of putative nucleic acid-dependent ATPases including MCM proteins involved in the initiation of eukaryotic DNA replication. Nucleic Acids Res. 1993 Jun 11;21(11):2541–2547. doi: 10.1093/nar/21.11.2541. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Larsen P. I., Sydnes L. K., Landfald B., Strøm A. R. Osmoregulation in Escherichia coli by accumulation of organic osmolytes: betaines, glutamic acid, and trehalose. Arch Microbiol. 1987 Feb;147(1):1–7. doi: 10.1007/BF00492896. [DOI] [PubMed] [Google Scholar]
- Lucht J. M., Bremer E. Adaptation of Escherichia coli to high osmolarity environments: osmoregulation of the high-affinity glycine betaine transport system proU. FEMS Microbiol Rev. 1994 May;14(1):3–20. doi: 10.1111/j.1574-6976.1994.tb00067.x. [DOI] [PubMed] [Google Scholar]
- Luque I., Herrero A., Flores E., Madueño F. Clustering of genes involved in nitrate assimilation in the cyanobacterium Synechococcus. Mol Gen Genet. 1992 Mar;232(1):7–11. doi: 10.1007/BF00299130. [DOI] [PubMed] [Google Scholar]
- Merchán F., Kindle K. L., Llama M. J., Serra J. L., Fernández E. Cloning and sequencing of the nitrate transport system from the thermophilic, filamentous cyanobacterium Phormidium laminosum: comparative analysis with the homologous system from Synechococcus sp. PCC 7942. Plant Mol Biol. 1995 Jul;28(4):759–766. doi: 10.1007/BF00021199. [DOI] [PubMed] [Google Scholar]
- Mikkat S., Hagemann M., Schoor A. Active transport of glucosylglycerol is involved in salt adaptation of the cyanobacterium Synechocystis sp. strain PCC 6803. Microbiology. 1996 Jul;142(Pt 7):1725–1732. doi: 10.1099/13500872-142-7-1725. [DOI] [PubMed] [Google Scholar]
- Omata T., Andriesse X., Hirano A. Identification and characterization of a gene cluster involved in nitrate transport in the cyanobacterium Synechococcus sp. PCC7942. Mol Gen Genet. 1993 Jan;236(2-3):193–202. doi: 10.1007/BF00277112. [DOI] [PubMed] [Google Scholar]
- Overduin P., Boos W., Tommassen J. Nucleotide sequence of the ugp genes of Escherichia coli K-12: homology to the maltose system. Mol Microbiol. 1988 Nov;2(6):767–775. doi: 10.1111/j.1365-2958.1988.tb00088.x. [DOI] [PubMed] [Google Scholar]
- Perroud B., Le Rudulier D. Glycine betaine transport in Escherichia coli: osmotic modulation. J Bacteriol. 1985 Jan;161(1):393–401. doi: 10.1128/jb.161.1.393-401.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reddy K. J., Webb R., Sherman L. A. Bacterial RNA isolation with one hour centrifugation in a table-top ultracentrifuge. Biotechniques. 1990 Mar;8(3):250–251. [PubMed] [Google Scholar]
- Strøm A. R., Kaasen I. Trehalose metabolism in Escherichia coli: stress protection and stress regulation of gene expression. Mol Microbiol. 1993 Apr;8(2):205–210. doi: 10.1111/j.1365-2958.1993.tb01564.x. [DOI] [PubMed] [Google Scholar]
- Umesono K., Inokuchi H., Shiki Y., Takeuchi M., Chang Z., Fukuzawa H., Kohchi T., Shirai H., Ohyama K., Ozeki H. Structure and organization of Marchantia polymorpha chloroplast genome. II. Gene organization of the large single copy region from rps'12 to atpB. J Mol Biol. 1988 Sep 20;203(2):299–331. doi: 10.1016/0022-2836(88)90002-2. [DOI] [PubMed] [Google Scholar]
- Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]