caa a22 4500 60550573X CHVBK 20210128100623.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s00253-015-6863-z doi (NATIONALLICENCE)springer-10.1007/s00253-015-6863-z Fine-tuning recA expression in Staphylococcus aureus for antimicrobial photoinactivation: importance of photo-induced DNA damage in the photoinactivation mechanism [Elektronische Daten] [Mariusz Grinholc, Aleksandra Rodziewicz, Katarzyna Forys, Aleksandra Rapacka-Zdonczyk, Anna Kawiak, Anna Domachowska, Grzegorz Golunski, Christiane Wolz, Lili Mesak, Karsten Becker, Krzysztof Bielawski] Bacterial cell envelope is generally accepted as the primary target for a photo-induced oxidative stress. It is plausible that DNA damage occurs during the antimicrobial photoinactivation. Here we investigate the correlation between DNA damage and photoinactivation by evaluating the level of RecA-based DNA repair system in Staphylococcus aureus. By using exogenous photosensitizers (new methylene blue (NMB), toluidine blue O (TBO), 5,10,15,20-tetrakis(1-methyl-4-pyridinio)porphyrin tetra(p-toluenesulfonate) (TMPyP), zinc phthalocyanine (ZnPc), Rose Bengal (RB)) and ALA-induced endogenous porphyrin-dependent blue light (405nm), several outcomes were observed: (i) an increase of DNA damage (from gel electrophoresis in DNA damage assay), (ii) an increase of recA expression (luminescence assay in recA-lux strain), and (iii) an increase of RecA protein level (Western blotting). When recA expression was repressed by novobiocin, or abolished by deleting the gene, S. aureus susceptibility towards photoinactivation was increased at approximately a hundred-fold. The absence of RecA increases DNA damage to yield bactericidal effect. In novobiocin-resistant mutant (gyrB), as opposed to wild type, neither RecA protein level nor cell's susceptibility was affected by photoinactivation (when novobiocin is present). This is to suggest that GyrB-dependent inhibition mediated recA repression. Therefore, we have established the role of RecA in DNA damage during photoinactivation. With the use of rifampicin mutation frequency and Ames tests, we demonstrated that photoinactivation did not increase S. aureus mutagenesis and potentially is not mutagenic toward eukaryotic cells. The results suggest that the treatment is considered safe. In conclusion, we provide an evidence that recA inhibitor may serve as therapeutic adjuvant for antimicrobial photoinactivation. Clinical relevance of our findings warrants further investigations. The Author(s), 2015 DNA damage nationallicence Endogenous porphyrins nationallicence Mutagenesis nationallicence Photoinactivation nationallicence RecA nationallicence Staphylococcus aureus nationallicence Grinholc Mariusz Laboratory of Molecular Diagnostics, Department of Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822, Gdansk, Poland aut Rodziewicz Aleksandra Laboratory of Molecular Diagnostics, Department of Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822, Gdansk, Poland aut Forys Katarzyna Laboratory of Molecular Diagnostics, Department of Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822, Gdansk, Poland aut Rapacka-Zdonczyk Aleksandra Laboratory of Molecular Diagnostics, Department of Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822, Gdansk, Poland aut Kawiak Anna Department of Biotechnology, Division of Plant Protection and Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822, Gdansk, Poland aut Domachowska Anna Department of Biotechnology, Division of Plant Protection and Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822, Gdansk, Poland aut Golunski Grzegorz Laboratory of Biophysics, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822, Gdansk, Poland aut Wolz Christiane Interfaculty Institute of Microbiology and Infection Medicine, University of Tubingen, Wlfriede-Aulhorn-Strasse 6, 72076, Tubingen, Germany aut Mesak Lili Outreach, Research Training and Minority Science Program, Ayala School of Biological Sciences, University of California, 333 Steinhaus Hall, 92697-2525, Irvine, CA, USA aut Becker Karsten Institute of Medical Microbiology, University Hospital Münster, Domagkstr. 10, 48149, Münster, Germany aut Bielawski Krzysztof Laboratory of Molecular Diagnostics, Department of Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822, Gdansk, Poland aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/21(2015-11-01), 9161-9176 0175-7598 99:21<9161 2015 99 253 https://doi.org/10.1007/s00253-015-6863-z text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6863-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Grinholc Mariusz Laboratory of Molecular Diagnostics, Department of Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822, Gdansk, Poland aut NATIONALLICENCE 700 1- Rodziewicz Aleksandra Laboratory of Molecular Diagnostics, Department of Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822, Gdansk, Poland aut NATIONALLICENCE 700 1- Forys Katarzyna Laboratory of Molecular Diagnostics, Department of Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822, Gdansk, Poland aut NATIONALLICENCE 700 1- Rapacka-Zdonczyk Aleksandra Laboratory of Molecular Diagnostics, Department of Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822, Gdansk, Poland aut NATIONALLICENCE 700 1- Kawiak Anna Department of Biotechnology, Division of Plant Protection and Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822, Gdansk, Poland aut NATIONALLICENCE 700 1- Domachowska Anna Department of Biotechnology, Division of Plant Protection and Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822, Gdansk, Poland aut NATIONALLICENCE 700 1- Golunski Grzegorz Laboratory of Biophysics, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822, Gdansk, Poland aut NATIONALLICENCE 700 1- Wolz Christiane Interfaculty Institute of Microbiology and Infection Medicine, University of Tubingen, Wlfriede-Aulhorn-Strasse 6, 72076, Tubingen, Germany aut NATIONALLICENCE 700 1- Mesak Lili Outreach, Research Training and Minority Science Program, Ayala School of Biological Sciences, University of California, 333 Steinhaus Hall, 92697-2525, Irvine, CA, USA aut NATIONALLICENCE 700 1- Becker Karsten Institute of Medical Microbiology, University Hospital Münster, Domagkstr. 10, 48149, Münster, Germany aut NATIONALLICENCE 700 1- Bielawski Krzysztof Laboratory of Molecular Diagnostics, Department of Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, 80-822, Gdansk, Poland aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/21(2015-11-01), 9161-9176 0175-7598 99:21<9161 2015 99 253