Proteus mirabilis glutathione S-transferase B1-1 is involved in protective mechanisms against oxidative and chemical stresses.
We investigated the effects of several xenobiotics, including antimicrobial agents and general stress factors such as starvation, heat and osmotic shock, on the modulation of expression of Proteus mirabilis glutathione S-transferase B1-1 (PmGST B1-1). The level of expression of PmGST B1-1 was established by both Western- and Northern-blot experiments. Our results show that several compounds can modulate expression of PmGST B1-1. The level of PmGST B1-1 increased when bacterial cells were exposed to a variety of stresses such as 1-chloro-2,4-dinitrobenzene, H(2)O(2), fosfomycin or tetracycline. A knock-out gst B gene was also constructed using the suicide vector pKNOCKlox-Ap. Successful inactivation of the wild-type gene was confirmed by PCR, DNA sequence analysis and Western blotting. Under normal culture conditions, this mutant was viable and displayed no significant phenotypic differences compared with the wild-type. However, viability tests revealed that the null mutant was more sensitive to oxidative stress in the form of H(2)O(2) and to several antimicrobial drugs when compared with the wild-type. These results suggest that PmGST B1-1 has an active role in the protection against oxidative stress generated by H(2)O(2) and it appears to be involved in the detoxification of antimicrobial agents.
Allocati
Nerino
N
Dipartimento di Scienze Biomediche, Università G. d'Annunzio, Via dei Vestini 31, I-66013 Chieti, Italy. n.allocati@dsb.unich.it
Favaloro
Bartolo
B
Masulli
Michele
M
Alexeyev
Mikhail F
MF
Di Ilio
Carmine
C
D001483:Base Sequence; D017931:DNA Primers; D005982:Glutathione Transferase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007700:Kinetics; D018384:Oxidative Stress / Q000502:physiology*; D016133:Polymerase Chain Reaction; D011513:Proteus mirabilis / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D014158:Transcription, Genetic; D015262:Xenobiotics / Q000494:pharmacology
D017931:DNA Primers; D012333:RNA, Messenger; D011994:Recombinant Proteins; D015262:Xenobiotics; D006861:Hydrogen Peroxide; D005982:Glutathione Transferase