Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Novel gene clusters involved in arsenite oxidation and resistance in two arsenite oxidizers: Achromobacter sp. SY8 and Pseudomonas sp. TS44


This study describes three gene clusters involved in arsenic redox transformation of two arsenite oxidizers: Achromobacter sp. SY8 and Pseudomonas sp. TS44. A 17.5-kb sequence containing the arsenite oxidase (aox) gene cluster (aoxX-aoxS-aoxR and aoxA-aoxB-aoxC-aoxD) was isolated from SY8 using a fosmid library approach. Similarly, a 14.6-kb sequence including the aox cluster (arsD-arsA-aoxA-aoxB) and the arsenic resistance (ars) gene cluster (arsC1-arsR-arsC2-ACR3-arsH-dual specificity phosphatase (DSP)-glyceraldehyde-3-phosphate dehydrogenase (GAPDH)-major facilitator superfamily (MFS)) was obtained from TS44 by inverse polymerase chain reaction (PCR). According to reverse transcription (RT) PCR experiments, SY8 aoxXSR and aoxABCD transcribed as two different transcripts in opposite directions, and TS44 aox and ars clusters transcribed as a single transcript in their respective cluster. All of these genes were found to be upregulated by the addition of arsenite [As(III)], arsenate [As(V)], and antimonite [Sb(III)], except that TS44 arsC1-arsR appeared to be expressed constitutively. The SY8 aox cluster was predicted to be regulated by a two-component signal transduction system and a potential regulatory model was proposed. The TS44 aox cluster is unusual since it contains structural genes only and arsDA in its upstream. The TS44 ars cluster includes several genes previously identified not associated with arsenic resistance or transformation. This study showed novel structures and arrangements of arsenic gene clusters associated with bacterial As(III) oxidation and As(V) reduction.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6


  1. Achour AR, Bauda P, Billard P (2007) Diversity of arsenite transporter genes from arsenic-resistant soil bacteria. Res Microbiol 158:128–137

  2. Cai L, Liu G, Rensing C, Wang G (2009) Genes involved in arsenic transformation and resistance associated with different levels of arsenic-contaminated soils. BMC Microbiol 9:4

  3. Chen Y, Rosen BP (1997) Metalloregulatory properties of the ArsD repressor. J Biol Chem 272:14257–14262

  4. Dey S, Rosen BP (1995) Dual mode of energy coupling by the oxyanion-translocating ArsB protein. J Bacteriol 177:385–389

  5. Duquesne K, Lieutaud A, Ratouchniak J, Muller D, Lett MC, Bonnefoy V (2008) Arsenite oxidation by a chemoautotrophic moderately acidophilic Thiomonas sp.: from the strain isolation to the gene study. Environmental microbiology 10:228–237

  6. Ellis PJ, Conrads T, Hille R, Kuhn P (2001) Crystal structure of the 100 kDa arsenite oxidase from Alcaligenes faecalis in two crystal forms at 1.64 angstrom and 2.03 angstrom. Structure 9:125–132

  7. Gihring TM, Druschel GK, McCleskey RB, Hamers RJ, Banfield JF (2001) Rapid arsenite oxidation by Thermus aquaticus and Thermus thermophilus: field and laboratory investigations. Environ Sci & Technol 35:3857–3862

  8. Hara MR, Agrawal N, Kim SF, Cascio MB, Fujimuro M, Ozeki Y, Takahashi M, Cheah JH, Tankou SK, Hester LD, Ferris CD, Hayward SD, Snyder SH, Sawa A (2005) S-nitrosylated GAPDH initiates apoptotic cell death by nuclear translocation following Siah1 binding. Nat Cell Biol 7:665–674

  9. Huang Y, Lemieux MJ, Song J, Auer M, Wang DN (2003) Structure and mechanism of the glycerol-3-phosphate transporter from Escherichia coli. Science 301:616–620

  10. Inskeep WP, Macur RE, Hamamura N, Warelow TP, Ward SA, Santini JM (2007) Detection, diversity and expression of aerobic bacterial arsenite oxidase genes. Environmental microbiology 9:934–943

  11. Kashyap DR, Botero LM, Franck WL, Hassett DJ, McDermott TR (2006) Complex regulation of arsenite oxidation in Agrobacterium tumefaciens. J Bacteriol 188:1081–1088

  12. Lehr CR, Kashyap DR, McDermott TR (2007) New insights into microbial oxidation of antimony and arsenic. Appl Environ Microbiol 73:2386–2389

  13. Lenoble V, Deluchat V, Serpaud B, Bollinger JC (2003) Arsenite oxidation and arsenate determination by the molybdene blue method. Talanta 61:267–276

  14. Li X, Krumholz LR (2007) Regulation of arsenate resistance in Desulfovibrio desulfuricans G20 by an arsRBCC operon and an arsC gene. J Bacteriol 189:3705–3711

  15. Lin YF, Walmsley AR, Rosen BP (2006) An arsenic metallochaperone for an arsenic detoxification pump. Proc Natl Acad Sci U S A 103:15617–15622

  16. Mukhopadhyay R, Rosen BP, Phung LT, Silver S (2002) Microbial arsenic: from geocycles to genes and enzymes. FEMS Microbiol Rev 26:311–325

  17. Muller D, Medigue C, Koechler S, Barbe V, Barakat M, Talla E, Bonnefoy V, Krin E, Arsene-Ploetze F, Carapito C, Chandler M, Cournoyer B, Cruveiller S, Dossat C, Duval S, Heymann M, Leize E, Lieutaud A, Lievremont D, Makita Y, Mangenot S, Nitschke W, Ortet P, Perdrial N, Schoepp B, Siguier P, Simeonova DD, Rouy Z, Segurens B, Turlin E, Vallenet D, Van Dorsselaer A, Weiss S, Weissenbach J, Lett MC, Danchin A, Bertin PN (2007) A tale of two oxidation states: bacterial colonization of arsenic-rich environments. PLoS Genet 3:518–530

  18. Ordonez E, Letek M, Valbuena N, Gil JA, Mateos LM (2005) Analysis of genes involved in arsenic resistance in Corynebacterium glutamicum ATCC 13032. Appl Environ Microbiol 71:6206–6215

  19. Oremland RS, Stolz JF, Hollibaugh JT (2004) The microbial arsenic cycle in Mono Lake, California. FEMS Microbiol Ecol 48:15–27

  20. Pao SS, Paulsen IT, Saier MH Jr (1998) Major facilitator superfamily. Microbiol Mol Biol Rev 62:1–34

  21. Philips SE, Taylor ML (1976) Oxidation of arsenite to arsenate by Alcaligenes faecalis. Appl Environ Microbiol 32:392–399

  22. Pils B, Schultz J (2004) Evolution of the multifunctional protein tyrosine phosphatase family. Mol Biol Evol 21:625–631

  23. Qin J, Rosen BP, Zhang Y, Wang G, Franke S, Rensing C (2006) Arsenic detoxification and evolution of trimethylarsine gas by a microbial arsenite S-adenosylmethionine methyltransferase. Proc Natl Acad Sci U S A 103:2075–2080

  24. Rosen BP (1999) Families of arsenic transporters. Trends Microbiol 7:207–212

  25. Rosen BP (2002) Biochemistry of arsenic detoxification. FEBS Lett 529:86–92

  26. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor: NY

  27. Santini JM, vanden Hoven RN (2004) Molybdenum-containing arsenite oxidase of the chemolithoautotrophic arsenite oxidizer NT-26. J Bacteriol 186:1614–1619

  28. Sato T, Kobayashi Y (1998) The ars operon in the skin element of Bacillus subtilis confers resistance to arsenate and arsenite. J Bacteriol 180:1655–1661

  29. Silver S, Phung LT (2005) Genes and enzymes involved in bacterial oxidation and reduction of inorganic arsenic. Appl Environ Microbiol 71:599–608

  30. Stock AM, Robinson VL, Goudreau PN (2000) Two-component signal transduction. Annu Rev Biochem 69:183–215

  31. Wang G, Kennedy SP, Fasiludeen S, Rensing C, DasSarma S (2004) Arsenic resistance in Halobacterium sp. strain NRC-1 examined by using an improved gene knockout system. J Bacteriol 186:3187–3194

  32. Weeger W, Lievremont D, Perret M, Lagarde F, Hubert JC, Leroy M, Lett MC (1999) Oxidation of arsenite to arsenate by a bacterium isolated from an aquatic environment. BioMetals 12:141–149

  33. Wu J, Rosen BP (1993) The arsD gene encodes a second trans-acting regulatory protein of the plasmid-encoded arsenical resistance operon. Mol Microbiol 8:615–623

  34. Zheng L, Roeder RG, Luo Y (2003) S phase activation of the histone H2B promoter by OCA-S, a coactivator complex that contains GAPDH as a key component. Cell 114:255–266

  35. Zhou Y (1990) Arsenite and arsenate determination by the molybdene blue method. Environmental Protection Science (Chinese) 16:45–47

  36. Zhou T, Radaev S, Rosen BP, Gatti DL (2000) Structure of the ArsA ATPase: the catalytic subunit of a heavy metal resistance pump. Embo J 19:4838–4845

Download references


This work was supported by the National Natural Science Foundation of China (30570058), the PhD Supervisor Fund (20060504027), and the Retuning Oversea Scientist Fund of the Ministry of Education of China.

Author information

Correspondence to Gejiao Wang.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Cai, L., Rensing, C., Li, X. et al. Novel gene clusters involved in arsenite oxidation and resistance in two arsenite oxidizers: Achromobacter sp. SY8 and Pseudomonas sp. TS44. Appl Microbiol Biotechnol 83, 715–725 (2009).

Download citation


  • Arsenic
  • Arsenite oxidizer
  • aox cluster
  • ars cluster