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J. Bacteriol. doi:10.1128/JB.00375-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Plasmids pMOL28 and pMOL30 of Cupriavidus metallidurans are specialised in the maximal viable response to heavy metals

Molecular & Cellular Biology, Institute for Health, Environment & Safety, SCK•CEN, Boeretang 200, B-2400 Mol, Belgium, Service de Conformation des Macromolécules Biologique et de Bioinformatique, Université Libre de Bruxelles, Belgium, INRA, Département de Microbiologie, F-78850 Thiverval Grignon, France, Brookhaven National Laboratory, Biology Department, Upton, NY 11973-5000, USA

^* To whom correspondence should be addressed. Email: mmergeay{at}sckcen.be.

	Abstract

We fully annotated two large plasmids, pMOL28 (164 ORFs; 171 459 bp) and pMOL30 (247 ORFs; 233 720 bp), in the genome of Cupriavidus metallidurans CH34. pMOL28 contains a backbone of maintenance- and transfer-genes resembling those found in the plasmids pSym of C. taiwanensis and pHG1 of C. eutrophus, suggesting they belong to a new class of plasmids. Genes involved in the resistance to the heavy metals Co(II), Cr(VI), Hg(II), and Ni(II) are concentrated within a region of 34kb on pMOL28; those involved in the resistance to Ag(I), Cd(II), Co(II), Cu(II), Hg(II), Pb(II), and Zn(II) occur within a 132kb region on pMOL30. We identified putative genomic islands containing metal-resistance operons flanked by mobile genetic elements, one on pMOL28 and two on pMOL30. Transcriptomic analysis using quantitative PCR and microarrays revealed the metal-mediated up-regulation of 83 genes on pMOL28 and of 143 genes on pMOL30 that coded for all known heavy-metal resistance proteins and some new ones (czcJ, mmrQ, and pbrU), membrane proteins, truncated transposases, conjugative transfer-proteins, and many unknown proteins. Five genes on each plasmid were down-regulated; for one of them, chrI localised on pMOL28, this occurs in the presence of five cations.

We observed multiple cross-responses (the induction of specific metal resistances by other metals), suggesting that the cellular defences of C. metallidurans against heavy metal stress involves various regulons, and probably has multiple stages including a more general response, and a more metal-specific one.

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