Aiming at developing cyanobacterial-based biosensors for heavy metal detection, expression of heavy metal inducible genes of the cyanobacterium Synechocystis PCC 6803 was investigated by quantitative RT-PCR upon 15 minutes exposure to biologically relevant concentrations of Co 2+ , Zn 2+ , Ni 2+ , Cd 2+ , Cr 6+ , As 3+ and As 5+ . The ziaA gene, which encodes a Zn 2+ -transporting P-type ATPase showed a markedly increased mRNA level after incubation with Cd 2+ and arsenic ions, besides the expected induction by Zn 2+ ions. The Co 2+ efflux system-encoding gene coaT was strongly induced by Co 2+ and Zn 2+ ions, moderately induced by As 3+ ions, and induced at a relatively low level by Cd 2+ and As 5+ ions. Expression of nrsB , which encodes a part of a putative Ni 2+ efflux system was highly induced by Ni 2+ salts and at a low extent by Co 2+ and Zn 2+ salts. The arsB gene, which encodes a putative arsenite-specific efflux pump was highly induced by As 3+ and As 5+ ions, while other metal salts provoked insignificant transcript level increase. The transcript of chrA , in spite of the high sequence similarity of its protein product with several bacterial chromate transporters, shows no induction upon Cr 6+ salt exposure. We conclude that due to the largely unspecific heavy metal response of the studied genes only nrsB and arsB are potential candidates for biosensing applications for detection of Ni 2+ and arsenic pollutants, respectively.
Blum, L. J., Gautier, S. M., Coulet, P. R. (1989) Design of luminescence photobiosensors. J. Biolumin. Chemilumin. 4 , 543–550.
Coulet P. R. , 'Design of luminescence photobiosensors ' (1989 ) 4 J. Biolumin. Chemilumin. : 543 -550 .
Cervantes, C., Ohtake, H., Chu, L., Misra, T. K., Silver, S. (1990) Cloning, nucleotide-sequence, and expression of the chromate resistance determinant of Pseudomonas-Aeruginosa plasmid Pum505. J. Bacteriol. 172 , 287–291.
Silver S. , 'Cloning, nucleotide-sequence, and expression of the chromate resistance determinant of Pseudomonas-Aeruginosa plasmid Pum505 ' (1990 ) 172 J. Bacteriol. : 287 -291 .
García-Domínguez, M., Lopez-Maury, L., Florencio, F. J., Reyes, J. C. (2000) A gene cluster involved in metal homeostasis in the cyanobacterium Synechocystis sp. strain PCC 6803. J. Bacteriol. 182 , 1507–1514.
Reyes J. C. , 'A gene cluster involved in metal homeostasis in the cyanobacterium Synechocystis sp. strain PCC 6803 ' (2000 ) 182 J. Bacteriol. : 1507 -1514 .
Kaneko, T., Tanaka, A., Sato, S., Kotani, H., Sazuka, T., Miyajima, N., Sugiura, M., Tabata, S. (1995) Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803. I. Sequence features in the 1 Mb region from map positions 64% to 92% of the genome. DNA Res. 2 , 153–166.
Tabata S. , 'Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803. I. Sequence features in the 1 Mb region from map positions 64% to 92% of the genome ' (1995 ) 2 DNA Res. : 153 -166 .
Kaneko, T., Sato, S., Kotani, H., Tanaka, A., Asamizu, E., Nakamura, Y., Miyajima, N., Hirosawa, M., Sugiura, M., Sasamoto, S., Kimura, T., Hosouchi, T., Matsuno, A., Muraki, A., Nakazaki, N., Naruo, K., Okumura, S., Shimpo, S., Takeuchi, C., Wada, T., Watanabe, A., Yamada, M., Yasuda, M., Tabata, S. (1996) Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. Strain PCC6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions. DNA Res. 3 , 109–136.
Tabata S. , 'Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. Strain PCC6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions ' (1996 ) 3 DNA Res. : 109 -136 .
Liao, V. H. C., Chien, M. T., Tseng, Y. Y., Ou, K. L. (2006) Assessment of heavy metal bioavailability in contaminated sediments and soils using green fluorescent protein-based bacterial biosensors. Environ. Pollut. 142 , 17–23.
Ou K. L. , 'Assessment of heavy metal bioavailability in contaminated sediments and soils using green fluorescent protein-based bacterial biosensors ' (2006 ) 142 Environ. Pollut. : 17 -23 .
Lopez-Maury, L., Florencio, F. J., Reyes, J. C. (2003) Arsenic sensing and resistance system in the cyanobacterium Synechocystis sp. strain PCC 6803. J. Bacteriol. 185 , 5363–5371.
Reyes J. C. , 'Arsenic sensing and resistance system in the cyanobacterium Synechocystis sp. strain PCC 6803 ' (2003 ) 185 J. Bacteriol. : 5363 -5371 .
Lopez-Maury, L., Garcia-Dominguez, M., Florencio, F. J., Reyes, J. C. (2002) A two-component signal transduction system involved in nickel sensing in the cyanobacterium Synechocystis sp. PCC 6803. Mol. Microbiol. 43 , 247–256.
Reyes J. C. , 'A two-component signal transduction system involved in nickel sensing in the cyanobacterium Synechocystis sp. PCC 6803 ' (2002 ) 43 Mol. Microbiol. : 247 -256 .
Mohamed, A., Janson, C. (1989) Influence of light on accumulation of photosynthesis-specific transcripts in the cyanobacterium Synechocystis 6803. Plant Mol. Biol. 13 , 693–700.
Janson C. , 'Influence of light on accumulation of photosynthesis-specific transcripts in the cyanobacterium Synechocystis 6803 ' (1989 ) 13 Plant Mol. Biol. : 693 -700 .
Mukhopadhyay, R., Rosen, B. P., Pung, L. T., Silver, S. (2002) Microbial arsenic: from geocycles to genes and enzymes. FEMS Microbiol. Rev. 26 , 311–325.
Silver S. , 'Microbial arsenic: from geocycles to genes and enzymes ' (2002 ) 26 FEMS Microbiol. Rev. : 311 -325 .
Neyt, C., Iriarte, M., Thi, V. H., Cornelis, G. R. (1997) Virulence and arsenic resistance in yersiniae. J. Bacteriol. 179 , 612–619.
Cornelis G. R. , 'Virulence and arsenic resistance in yersiniae ' (1997 ) 179 J. Bacteriol. : 612 -619 .
Nicholson, M. L., Laudenbach, D. E. (1995) Genes encoded on a cyanobacterial plasmid are transcriptionally regulated by sulfur availability and cysR. J. Bacteriol. 177 , 2143–2150.
Laudenbach D. E. , 'Genes encoded on a cyanobacterial plasmid are transcriptionally regulated by sulfur availability and cysR ' (1995 ) 177 J. Bacteriol. : 2143 -2150 .
Nies, A., Nies, D. H., Silver, S. (1990) Nucleotide sequence and expression of a plasmid-encoded chromate resistance determinant from Alcaligenes eutrophus . J. Biol. Chem. 265 , 5648–5653.
Silver S. , 'Nucleotide sequence and expression of a plasmid-encoded chromate resistance determinant from Alcaligenes eutrophus ' (1990 ) 265 J. Biol. Chem. : 5648 -5653 .
Nies, D. H., Koch, S., Wachi, S., Peitzsch, N., Saier, M. H. (1998) CHR, a novel family of prokaryotic proton motive force-driven transporters probably containing chromate/sulfate antiporters. J. Bacteriol. 180 , 5799–5802.
Saier M. H. , 'CHR, a novel family of prokaryotic proton motive force-driven transporters probably containing chromate/sulfate antiporters ' (1998 ) 180 J. Bacteriol. : 5799 -5802 .
Nies, D. H., Nies, A., Chu, L., Silver, S. (1989) Expression and nucleotide sequence of a plasmid-determined divalent cation efflux system from Alcaligenes eutrophus . Proc. Natl. Acad. Sci. USA 86 , 7351–7355.
Silver S. , 'Expression and nucleotide sequence of a plasmid-determined divalent cation efflux system from Alcaligenes eutrophus ' (1989 ) 86 Proc. Natl. Acad. Sci. USA : 7351 -7355 .
Nucifora, G., Chu, L., Misra, T. K., Silver, S. (1989) Cadmium resistance from Staphylococcus aureus plasmid pI258 cadA gene results from a cadmium-efflux ATPase. Proc. Natl. Acad. Sci. USA 86 , 3544–3548.
Silver S. , 'Cadmium resistance from Staphylococcus aureus plasmid pI258 cadA gene results from a cadmium-efflux ATPase ' (1989 ) 86 Proc. Natl. Acad. Sci. USA : 3544 -3548 .
Ramanathan, S., Shi, W. P., Rosen, B. P., Daunert, S. (1997) Sensing antimonite and arsenite at the subattomole level with genetically engineered bioluminescent bacteria. Anal. Chem. 69 , 3380–3384.
Daunert S. , 'Sensing antimonite and arsenite at the subattomole level with genetically engineered bioluminescent bacteria ' (1997 ) 69 Anal. Chem. : 3380 -3384 .
Rippka, R., Deruelles, J., Waterbury, J. B., Herdman, M., Stanier, R. Y. (1979) Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J. Gen. Microbiol. 111 , 1–61.
Stanier R. Y. , 'Generic assignments, strain histories and properties of pure cultures of cyanobacteria ' (1979 ) 111 J. Gen. Microbiol. : 1 -61 .
Rosen, B. P. (2002) Biochemistry of arsenic detoxification. FEBS Lett. 529 , 86–92.
Rosen B. P. , 'Biochemistry of arsenic detoxification ' (2002 ) 529 FEBS Lett. : 86 -92 .
Rutherford, J. C., Cavet, J. S., Robinson, N. J. (1999) Cobalt-dependent transcriptional switching by a dual-effector MerR-like protein regulates a cobalt-exporting variant CPx-type ATPase. J. Biol. Chem. 274 , 25827–25832.
Robinson N. J. , 'Cobalt-dependent transcriptional switching by a dual-effector MerR-like protein regulates a cobalt-exporting variant CPx-type ATPase ' (1999 ) 274 J. Biol. Chem. : 25827 -25832 .
Shi, W. P., Wu, J. H., Rosen, B. P. (1994) Identification of a putative metal-binding site in a new family of metalloregulatory proteins. J. Biol. Chem. 269 , 19826–19829.
Rosen B. P. , 'Identification of a putative metal-binding site in a new family of metalloregulatory proteins ' (1994 ) 269 J. Biol. Chem. : 19826 -19829 .
Silver, S., Phung, L. T. (1996) Bacterial heavy metal resistance: New surprises. Annu. Rev. Microbiol. 50 , 753–789.
Phung L. T. , 'Bacterial heavy metal resistance: New surprises ' (1996 ) 50 Annu. Rev. Microbiol. : 753 -789 .
Silver, S., Phung, L. T. (2005) Genes and enzymes involved in bacterial oxidation and reduction of inorganic arsenic. Appl. Environ. Microbiol. 71 , 599–608.
Phung L. T. , 'Genes and enzymes involved in bacterial oxidation and reduction of inorganic arsenic ' (2005 ) 71 Appl. Environ. Microbiol. : 599 -608 .
Tauriainen, S., Karp, H., Chang, W., Virta, M. (1997) Recombinant luminescent bacteria for measuring bioavailable arsenite and antimonite. Appl. Environ. Microbiol. 63 , 4456–4461.
Virta M. , 'Recombinant luminescent bacteria for measuring bioavailable arsenite and antimonite ' (1997 ) 63 Appl. Environ. Microbiol. : 4456 -4461 .
Thelwell, C., Robinson, N. J., Turner-Cavet, J. S. (1998) An SmtB-like repressor from Synechocystis PCC 6803 regulates a zinc exporter. Proc. Natl. Acad. Sci. USA 95 , 10728–10733.
Turner-Cavet J. S. , 'An SmtB-like repressor from Synechocystis PCC 6803 regulates a zinc exporter ' (1998 ) 95 Proc. Natl. Acad. Sci. USA : 10728 -10733 .
Tong, L., Nakashima, S., Shibasaka, M., Katsuhara, M., Kasamo, K. (2002) A novel histidine-rich CPx-ATPase from the filamentous cyanobacterium Oscillatoria brevis related to multiple-heavymetal cotolerance. J. Bacteriol. 184 , 5027–5035.
Kasamo K. , 'A novel histidine-rich CPx-ATPase from the filamentous cyanobacterium Oscillatoria brevis related to multiple-heavymetal cotolerance ' (2002 ) 184 J. Bacteriol. : 5027 -5035 .
Trevors, J. T., Stratton, G. W., Gadd, G. M. (1986) Cadmium transport, resistance, and toxicity in bacteria, algae, and fungi. Can. J. Microbiol. 32 , 447–464.
Gadd G. M. , 'Cadmium transport, resistance, and toxicity in bacteria, algae, and fungi ' (1986 ) 32 Can. J. Microbiol. : 447 -464 .
Turner, J. S., Robinson, N. J. (1995) Cyanobacterial metallothioneins-biochemistry and molecular genetics. J. Ind. Microbiol. 14 , 119–125.
Robinson N. J. , 'Cyanobacterial metallothioneins-biochemistry and molecular genetics ' (1995 ) 14 J. Ind. Microbiol. : 119 -125 .
Wysocki, R., Bobrowicz, P., Ulaszewski, S. (1997) The Saccharomyces cerevisiae ACR3 gene encodes a putative membrane protein involved in arsenite transport. J. Biol. Chem. 272 , 30061–30066.
Ulaszewski S. , 'The Saccharomyces cerevisiae ACR3 gene encodes a putative membrane protein involved in arsenite transport ' (1997 ) 272 J. Biol. Chem. : 30061 -30066 .