title: Superoxide dismutase is upregulated in Staphylococcus aureus following protoporphyrin-mediated photodynamic inactivation and does not directly influence the response to photodynamic treatment.
abstract: BACKGROUND
  Staphylococcus aureus, a major human pathogen causes a wide range of disease syndromes. The most dangerous are methicillin-resistant S. aureus (MRSA) strains, resistant not only to all β-lactam antibiotics but also to other antimicrobials. An alarming increase in antibiotic resistance spreading among pathogenic bacteria inclines to search for alternative therapeutic options, for which resistance can not be developed easily. Among others, photodynamic inactivation (PDI) of S. aureus is a promising option. Photodynamic inactivation is based on a concept that a non toxic chemical, called a photosensitizer upon excitation with light of an appropriate wavelength is activated. As a consequence singlet oxygen and other reactive oxygen species (e.g. superoxide anion) are produced, which are responsible for the cytotoxic effect towards bacterial cells. As strain-dependence in photodynamic inactivation of S. aureus was observed, determination of the molecular marker(s) underlying the mechanism of the bacterial response to PDI treatment would be of great clinical importance. We examined the role of superoxide dismutases (Sod) in photodynamic inactivation of S. aureus as enzymes responsible for oxidative stress resistance.
  
  
  RESULTS
  The effectiveness of photodynamic inactivation towards S. aureus and its Sod isogenic mutants deprived of either of the two superoxide dismutase activities, namely SodA or SodM or both of them showed similar results, regardless of the Sod status in TSB medium. On the contrary, in the CL medium (without Mn++ ions) the double SodAM mutant was highly susceptible to photodynamic inactivation. Among 8 clinical isolates of S. aureus analyzed (4 MRSA and 4 MSSA), strains highly resistant and strains highly vulnerable to photodynamic inactivation were noticed. We observed that Sod activity as well as sodA and sodM transcript level increases after protoporphyrin IX-based photodynamic treatment but only in PDI-sensitive strains.
  
  
  CONCLUSIONS
  We confirmed that porphyrin-based photokilling efficacy is a strain-dependent phenomenon. We showed that oxidative stress sensitivity caused by the lack of both Sod enzymes can be relieved in the presence of Mn ions and partially in the presence of Fe ions. The fact that Sod activity increase is observed only in PDI-susceptible cells emphasizes that this is probably not a direct factor affecting S. aureus vulnerability to porphyrin-based PDI.
authors.affiliation: Intercollegiate Faculty of Biotechnology University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822 Gdansk, Poland. strzala@biotech.ug.gda.pl
authors.forename: Joanna
authors.initials: J
authors.lastname: Nakonieczna
authors.forename: Ewelina
authors.initials: E
authors.lastname: Michta
authors.forename: Magda
authors.initials: M
authors.lastname: Rybicka
authors.forename: Mariusz
authors.initials: M
authors.lastname: Grinholc
authors.forename: Anna
authors.initials: A
authors.lastname: Gwizdek-Wiśniewska
authors.forename: Krzysztof P
authors.initials: KP
authors.lastname: Bielawski
mesh_terms: D000900:Anti-Bacterial Agents / Q000378:metabolism*; D055786:Gene Knockout Techniques; D006801:Humans; D007501:Iron / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D017319:Photosensitizing Agents / Q000378:metabolism*; D011524:Protoporphyrins / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013211:Staphylococcus aureus / Q000187:drug effects* / Q000201:enzymology*; D013482:Superoxide Dismutase / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D000900:Anti-Bacterial Agents; D017319:Photosensitizing Agents; D011524:Protoporphyrins; D017382:Reactive Oxygen Species; D008345:Manganese; D007501:Iron; D013482:Superoxide Dismutase