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Perinuclear formation of Reactive Nitrogen Species causes DNA damage in Methicillin‐resistant Staphylococcus Aureus induced sepsis.
Author(s) -
Bansal Kamna,
Connelly Rhykka L,
Herndon David,
Traber Daniel L,
Enkhbaatar Perenlei
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.1003.11
Subject(s) - nitrotyrosine , dna damage , blot , nadph oxidase , western blot , staphylococcus aureus , reactive nitrogen species , methicillin resistant staphylococcus aureus , microbiology and biotechnology , confocal , sepsis , nitric oxide , programmed cell death , chemistry , biology , nitric oxide synthase , reactive oxygen species , apoptosis , dna , immunology , bacteria , biochemistry , gene , genetics , geometry , mathematics , organic chemistry
Rationale MRSA commonly referred to as a 'super bug' because of its resistance to antibiotic therapy, is now a common infection encountered in hospitals and communities. We have previously shown that MRSA causes vascular hyperpermeability, multi‐organ system failure, and death in our ovine model of MRSA‐induced sepsis. We hypothesized that RNS may play a pivotal role in MRSA‐induced DNA damage and consequent cellular death. Methods Pulmonary epithelial cell (A549) were challenged with 105 cfu MRSA over a time course of 4 hrs then probed for markers of RNS (2,7‐Dichlorodihydrofluorescein [DCF] and 3‐Nitrotyrosine {3‐NT}), as well as poly ADP‐ribose (PAR), a marker of DNA damage, by confocal imaging and western blot analyses. Neuronal nitric oxide synthase (nNOS) and NADPH oxidase‐4 (NOX4) were localized by confocal microscopy. Results MRSA caused nNOS and NOX4 translocation to a perinuclear location, resulting in a 7‐fold increase in RNS (measured by DCF) at the perinuclear compartment compared to normal cells. MRSA also caused PAR (3.4 fold) and 3‐NT (4.8 fold) formation within 1h of MRSA exposure observed by both confocal and western blotting analyses. Conclusions MRSA causes DNA damage in alveolar epithelial cells as result of perinuclear formation of RNS .