1887

Abstract

sp. strain CNB-1, a chloronitrobenzene-degrading bacterium, was demonstrated to possess higher arsenate tolerance as compared with the mutant strain CNB-2. pCNB1, a plasmid harboured by CNB-1 but not CNB-2, contained the genetic cluster , which putatively encodes arsenate-resistance regulator, family II arsenate reductase, arsenite efflux pump and family I arsenate reductase, respectively, in strain CNB-1. The -negative could gain arsenate resistance by transformation with or , indicating that these two genes might express functional forms of arsenate reductases. Intriguingly, when CNB-1 cells were exposed to arsenate, the transcription of and was measurable by RT-PCR, but only ArsP was detectable at protein level. To explore the proteins responding to arsenate stress, CNB-1 cells were cultured with and without arsenate and differential proteomics was carried out by two-dimensional PAGE (2-DE) and MALDI-TOF MS. A total of 31 differential 2-DE spots were defined upon image analysis and 23 proteins were identified to be responsive specifically to arsenate. Of these spots, 18 were unique proteins. These proteins were identified to be phosphate transporters, heat-shock proteins involved in protein refolding, and enzymes participating in carbon and energy metabolism.

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2007-11-01
2019-10-18
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Phylogenetic analysis of functionally identified (shown in bold) and putative prokaryotic arsenate reductases. The phylogenetic tree was generated using the neighbour-joining method with the Mega 4.0 software. The scale at the bottom shows sequence divergence. Numbers at nodes show the support from 1000 bootstraps. The eukaryotic arsenate reductases from spp. were used as outgroup. [ PDF] (75 kb) Comparative proteomes of sp. strain CNB-1 grown in the presence (top) and the absence (bottom) of arsenate. CNB-1 was cultured in LB broth with and without 100 mM sodium arsenate for 24 h. 2-DE was performed using different pH gradient strips (pH 4-7 and pH 6-11). Numbered proteins were differentially expressed under arsenate stress, and their identities are listed in Table 2 of the main paper. The proteins that showed increased abundance or only appeared in the presence of arsenate are marked with solid arrows (upper panels), and the proteins that showed increased abundance or only appeared in the absence of arsenate are marked with open arrows (lower panels). [ PDF] (1.4 Mb) Global responses to arsenate exposure in sp. strain CNB-1. Up-regulated proteins are marked with triangles; induced proteins are marked with squares. For further explanation of the figure, see the main paper. [ PDF] (66 kb)

PDF

Phylogenetic analysis of functionally identified (shown in bold) and putative prokaryotic arsenate reductases. The phylogenetic tree was generated using the neighbour-joining method with the Mega 4.0 software. The scale at the bottom shows sequence divergence. Numbers at nodes show the support from 1000 bootstraps. The eukaryotic arsenate reductases from spp. were used as outgroup. [ PDF] (75 kb) Comparative proteomes of sp. strain CNB-1 grown in the presence (top) and the absence (bottom) of arsenate. CNB-1 was cultured in LB broth with and without 100 mM sodium arsenate for 24 h. 2-DE was performed using different pH gradient strips (pH 4-7 and pH 6-11). Numbered proteins were differentially expressed under arsenate stress, and their identities are listed in Table 2 of the main paper. The proteins that showed increased abundance or only appeared in the presence of arsenate are marked with solid arrows (upper panels), and the proteins that showed increased abundance or only appeared in the absence of arsenate are marked with open arrows (lower panels). [ PDF] (1.4 Mb) Global responses to arsenate exposure in sp. strain CNB-1. Up-regulated proteins are marked with triangles; induced proteins are marked with squares. For further explanation of the figure, see the main paper. [ PDF] (66 kb)

PDF

Phylogenetic analysis of functionally identified (shown in bold) and putative prokaryotic arsenate reductases. The phylogenetic tree was generated using the neighbour-joining method with the Mega 4.0 software. The scale at the bottom shows sequence divergence. Numbers at nodes show the support from 1000 bootstraps. The eukaryotic arsenate reductases from spp. were used as outgroup. [ PDF] (75 kb) Comparative proteomes of sp. strain CNB-1 grown in the presence (top) and the absence (bottom) of arsenate. CNB-1 was cultured in LB broth with and without 100 mM sodium arsenate for 24 h. 2-DE was performed using different pH gradient strips (pH 4-7 and pH 6-11). Numbered proteins were differentially expressed under arsenate stress, and their identities are listed in Table 2 of the main paper. The proteins that showed increased abundance or only appeared in the presence of arsenate are marked with solid arrows (upper panels), and the proteins that showed increased abundance or only appeared in the absence of arsenate are marked with open arrows (lower panels). [ PDF] (1.4 Mb) Global responses to arsenate exposure in sp. strain CNB-1. Up-regulated proteins are marked with triangles; induced proteins are marked with squares. For further explanation of the figure, see the main paper. [ PDF] (66 kb)

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