title: Identification of an archaeal mercury regulon by chromatin immunoprecipitation.
abstract: Mercury is a heavy metal and toxic to all forms of life. Metal exposure can invoke a response to improve survival. In archaea, several components of a mercury response system have been identified, but it is not known whether metal transport is a member of this system. To identify such missing components, a peptide-tagged MerR transcription factor was used to localize enriched chromosome regions by chromosome immunoprecipitation combined with DNA sequence analysis. Such regions could serve as secondary regulatory binding sites to control the expression of additional genes associated with mercury detoxification. Among the 31 highly enriched loci, a subset of five was pursued as potential candidates based on their current annotations. Quantitative reverse transcription-PCR analysis of these regions with and without mercury treatment in WT and mutant strains lacking merR indicated significant regulatory responses under these conditions. Of these, a Family 5 extracellular solute-binding protein and the MarR transcription factor shown previously to control responses to oxidation were most strongly affected. Inactivation of the solute-binding protein by gene disruption increased the resistance of mutant cells to mercury challenge. Inductively coupled plasma-MS analysis of the mutant cell line following metal challenge indicated there was less intracellular mercury compared with the isogenic WT strain. Together, these regulated genes comprise new members of the archaeal MerR regulon and reveal a cascade of transcriptional control not previously demonstrated in this model organism.
authors.affiliation: 1​ School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, Nebraska, USA.
authors.forename: Deepak
authors.initials: D
authors.lastname: Rudrappa
authors.affiliation: 3​ Department of Biomedical Engineering and Genome Center, University of California-Davis, Davis, California, USA.
authors.forename: Andrew I
authors.initials: AI
authors.lastname: Yao
authors.affiliation: 1​ School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, Nebraska, USA.
authors.forename: Derrick
authors.initials: D
authors.lastname: White
authors.affiliation: 2​ Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, Lincoln, Nebraska, USA.
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Pavlik
authors.affiliation: 1​ School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, Nebraska, USA.
authors.forename: Raghuveer
authors.initials: R
authors.lastname: Singh
authors.affiliation: 3​ Department of Biomedical Engineering and Genome Center, University of California-Davis, Davis, California, USA.
authors.forename: Marc T
authors.initials: MT
authors.lastname: Facciotti
authors.affiliation: 1​ School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, Nebraska, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Blum
mesh_terms: D001105:Archaea / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D047369:Chromatin Immunoprecipitation; D015964:Gene Expression Regulation, Bacterial; D008628:Mercury / Q000378:metabolism*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D018085:Regulon
chemical_list: D019843:Archaeal Proteins; D008628:Mercury