Abstract
A stable DNA/protein complex having an apparent molecular mass of approximately 150kDa was purified from nitrate-limited cultures of the cyanobacterium Synechococcus sp. strain PCC 7942. Amino-terminal peptide sequencing indicated that the polypeptide was structurally similar to the Dps protein of Escherichia coli; Dps is also known as the product of the starvation- and stationary-phase-inducible gene, pexB. The 150-kDa complex dissociated into a 22-kDa protein monomer after boiling in 2% SDS. The 150-kDa complex preparation had approximately a 10% nucleic acid content and upon dissociation released DNA fragments that were sensitive to S1 nuclease digestion. Immunoblot data indicated that the complex accumulates during stationary phase and during nitrogen, sulfur, and phosphorus limitation. DNA-binding assays indicated that the protein nonspecifically binds both linear and supercoiled DNA. Circular dichroism spectroscopy revealed that the Synechococcus sp. Dps-like protein contains extensive regions of alpha-helical secondary structure. We propose that the 150-kDa complex represents a hexameric aggregate of the Dps-like protein complexed with single-stranded DNA and serves to bind a portion of the chromosomal DNA under nutrient-limited conditions.
Similar content being viewed by others
References
Allen MM (1968) Simple conditions for growth of unicellular blue-green algae on plates. J Phycol 4:1–4
Almiron M, Link AJ, Furlong D, Kolter R (1992) A novel DNA-binding protein with regulatory and protective roles in starved Escherichia coli. Genes Dev 6:2646–2654
Altuvia S, Almiron M, Huisman G, Kolter R, Storz G (1994) The dps promoter is activated by OxyR during growth and by IHF and σs in stationary phase. Mol Microbiol 13:265–272
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Burkhart W (1993) In situ proteolytic digestions performed on proteins bound to the Hewlett-Packard hydrophobic sequencing column. In: Angeletti R (ed) Techniques in protein chemistry, vol 4. Academic Press, New York London, pp 399–406
Chen L, James LP, Helmann JD (1993) Metalloregulation in Bacillus subtilis: isolation and characterization of two genes differentially repressed by metal ions. J Bacteriol 175:5428–5437
Collier JL, Grossman AR (1994) A small polypeptide triggers complete degradation of light-harvesting phycobiliproteins in nutrient-deprived cyanobacteria. EMBO J 13:1039–1047
Copeland RA (1994) Methods for protein analysis, 1st edn. Chapman & Hall, London
Flores E, Guerrero MG, Losada M (1983) Photosynthetic nature of nitrate uptake and reduction in the cyanobacterium Anacystis nidulans. Biochim Biophys Acta 722:408–416
Foulds IJ, Carr NG (1977) A proteolytic enzyme degrading phycocyanin in the cyanobacterium Anabaena cylindrica. FEMS Microbiol Lett 2:117–119
Green LS, Grossman A (1988) Changes in sulfate transport characteristics and protein composition of Anacystis nidulans R2 during sulfur deprivation. J Bacteriol 170:583–587
Grossman AR, Schaefer MR, Chiang GG, Collier JL (1993) The phycobilisome, a light-harvesting complex responsive to environmental conditions. Microbiol Rev 57:725–749
Guikema J, Sherman LA (1981) Electrophoretic profiles of cyanobacterial membrane polypeptides showing heme-dependent peroxidase activity. Biochim Biophys Acta 637:189–201
Hartford OM, Dowds BCA (1994) Isolation and characterization of a hydrogen peroxide resistant mutant of Bacillus subtilis. Microbiology 140:297–304
Kulkarni RD, Schaefer MR, Golden SS (1992) Transcriptional and posttranscriptional components of psbA: response to high light intensity in Synechococcus sp. strain PCC 7942. J Bacteriol 174:3775–3781
Lau RH, MacKenzie MM, Doolittle WF (1977) Phycocyanin synthesis and degradation in the blue-green bacterium Anacystis nidulans. J Bacteriol 132:771–778
Lipman DJ, Pearson WR (1985) Rapid and sensitive protein similarity searches. Science 227:1435–1440
Lomovskaya OL, Kidwell JP, Matin A (1994) Characterization of the σ38-dependent expression of a core Escherichia coli starvation gene, pexB. J Bacteriol 176:3928–3935
Matin A (1991) The molecular basis of carbon-starvation-induced general resistance in Escherichia coli. Mol Microbiol 5:3–10
Melgar E, Goldthwait DA (1968) Deoxyribonucleic acid nucleases. 2. The effects of metals on the mechanism of action of deoxyribonuclease I. J Biol Chem 243:4409–4417
Mittler R, Tel-Or E (1991) Oxidative stress responses and shock proteins in the unicellular cyanobacterium Synechococcus R2 (PCC 7942). Arch Microbiol 155:125–130
Neidhardt FC (1987) Chemical composition of Escherichia coli. In: Neidhardt FC (ed) Escherichia coli and Salmonella typhimurium. Am Soc Microbiol, Washington DC, pp 3–6
Noordhoek GT, Hermans PWM, Paul AN, Schouls LM, Sluis JJ van der, Embden JDA van (1989) Treponema pallidum subspecies pallidum (Nichols) and Treponema pallidum subspecies pertenue (CDC 2575) differ in at least one nucleotide: comparion of two homologous antigens. Microb Pathog 6: 29–42
Pfeifer O, Pelletier I, Altenbuchner J, Van Pee KH (1992) Molecular cloning and sequencing of a non-haem bromoperoxidase gene from Streptomyces aureofaciens ATCC 10762. J Gen Microbiol 138:1123–1131
Reddy KJ, Bullerjahn GS, Sherman DM, Sherman LA (1988) Cloning, nucleotide sequencing and mutagenesis of a gene (irpA) involved in iron-deficient growth of the cyanobacterium Synechococcus sp. strain PCC 7942. J Bacteriol 170:4466–4476
Reuter W, Muller C (1993) Adaptation of the photosynthetic apparatus of cyanobacteria to light and CO2. J Photochem Photobiol B 21:3–27
Reyes JC, Florencio FJ (1994) A new type of glutamine synthetase in cyanobacteria: the protein encoded by the glnN gene supports nitrogen assimilation in Synechocystis sp. strain PCC 6803. J Bacteriol 176:1260–1267
Riethman HC, Bullerjahn G, Reddy KJ, Sherman LA (1988) Regulation of cyanobacterial pigment-protein composition and organization by environmental factors. Photosynth Res 18:131–161
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual. 2nd edn, Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Suzuki I, Sugiyama T, Omata T (1993) Primary structure and transcriptional regulation of the gene for nitrite reductase from the cyanobacterium Synechococcus PCC 7942. Plant Cell Physiol 34:1311–1320
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Vega-Palas MA, Madueño F, Herrero A, Flores E (1990) Identification and cloning of a regulatory gene for nitrogen assimilation in the cyanobacterium Synechococcus sp. strain PCC 7942. J Bacteriol 172:643–647
Webb R, Reddy KJ, Sherman LA (1990) Regulation and sequence of the Synechococcus sp. strain PCC 7942 groESL operon, encoding a cyanobacterial chaperonin. J Bacteriol 172:5079–5088
Wray W, Boulikas T, Wray VP, Hancock R (1981) Silver-staining of proteins in polyacrylamide gels. Anal Biochem 118:197–203
Yamanaka G, Glazer AN (1980) Dynamic aspects of phycobilisome structure. Phycobilisome turnover during nitrogen starvation in Synechococcus sp. Arch Microbiol 124:39–47
Yang JT, Wu CSC, Martinez HM (1986) Calculation of protein conformation from circular dichroism. Methods Enzymol 130: 228–236
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Peña, M.M.O., Burkhart, W. & Bullerjahn, G.S. Purification and characterization of a Synechococcus sp. strain PCC 7942 polypeptide structurally similar to the stress-induced Dps/PexB protein of Escherichia coli . Arch. Microbiol. 163, 337–344 (1995). https://doi.org/10.1007/BF00404206
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00404206