title: Characterization of the effects of an rpoC mutation that confers resistance to the Fst peptide toxin-antitoxin system toxin.
abstract: Overexpression of the Fst toxin in Enterococcus faecalis strain OG1X leads to defects in chromosome segregation, cell division and, eventually, membrane integrity. The M7 mutant derivative of OG1X is resistant to most of these effects but shows a slight growth defect in the absence of Fst. Full-genome sequencing revealed two differences between M7 and its OG1X parent. First, OG1X contains a frameshift mutation that inactivates the etaR response regulator gene, while M7 is a wild-type revertant for etaR. Second, the M7 mutant contains a missense mutation in the rpoC gene, which encodes the β' subunit of RNA polymerase. Mutagenesis experiments revealed that the rpoC mutation was primarily responsible for the resistance phenotype. Microarray analysis revealed that a number of transporters were induced in OG1X when Fst was overexpressed. These transporters were not induced in M7 in response to Fst, and further experiments indicated that this had a direct protective effect on the mutant cells. Therefore, exposure of cells to Fst appears to have a cascading effect, first causing membrane stress and then potentiation of these effects by overexpression of certain transporters.
authors.affiliation: Division of Basic Biomedical Sciences, Sanford School of Medicine, University of South Dakota, Vermillion, SD, USA.
authors.forename: Cassandra L
authors.initials: CL
authors.lastname: Brinkman
authors.forename: Roger
authors.initials: R
authors.lastname: Bumgarner
authors.forename: Weerayuth
authors.initials: W
authors.lastname: Kittichotirat
authors.forename: Paul M
authors.initials: PM
authors.lastname: Dunman
authors.forename: Lisa J
authors.initials: LJ
authors.lastname: Kuechenmeister
authors.forename: Keith E
authors.initials: KE
authors.lastname: Weaver
mesh_terms: D000483:Alleles; D000992:Antitoxins / Q000235:genetics / Q000378:metabolism*; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D004269:DNA, Bacterial; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism*; D013293:Enterococcus faecalis / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D009154:Mutation; D016133:Polymerase Chain Reaction
chemical_list: D000992:Antitoxins; D001427:Bacterial Toxins; D002352:Carrier Proteins; D004269:DNA, Bacterial; C473319:fst toxin, Enterococcus faecalis; C108570:beta' subunit of RNA polymerase; D012321:DNA-Directed RNA Polymerases