Bacterial peptide methionine sulphoxide reductase: co-induction with glutathione S-transferase during chemical stress conditions

Abstract

Peptide methionine sulphoxide reductase (MsrA; EC 1.8.4.6) is a ubiquitous enzyme catalysing the reduction of methionine sulphoxide to methionine in proteins, while the glutathione S-transferases (GSTs) are a major family of detoxification enzymes. A gene homologous to MsrA was identified in a chromosomal fragment from the bacterium Ochrobactrum anthropi, and this gene is located just downstream of a GST gene identified previously (OaGST) [Favaloro, Tamburro, Angelucci, De Luca, Melino, Di Ilio and Rotilio (1998) Biochem. J. 335, 573-579]. This raises the question of whether the products of these two genes may be involved in a common cellular protection function. To test this hypothesis, the hypothetical MsrA protein has been overexpressed in Escherichia coli as a functional 51 kDa GST fusion protein. Following cleavage with thrombin and purification, the soluble 24 kDa protein showed MsrA activity with N-acetylmethionine sulphoxide as substrate, as well as with other sulphoxide compounds. Therefore polyclonal antibodies were raised against the recombinant protein, and the modulation of MsrA in this bacterium, grown in the presence of different stimulants simulating several stress conditions, was investigated. The level of expression of MsrA was detected both by measuring the mRNA level and by immunoblotting experiments, in addition to measuring its catalytic activity. MsrA is a constitutive enzyme which is also inducible by chemical stress involving phenolic compounds such as phenol and 4-chlorophenol. Recently we reported that the GST of this bacterium, like MsrA, is only modulated by toxic chemical compounds [Favaloro, Tamburro, Trofino, Bologna, Rotilio and Heipieper (2000) Biochem. J. 346, 553-559]; therefore this is the first indication of a co-induction of the MsrA and GST enzymes during chemical stress.

Important Role for Methionine Sulfoxide Reductase in the Oxidative Stress Response of Xanthomonas campestris pv. phaseoli

“…NEM induction of msrA probably results from the depletion of thiol antioxidant molecules and the inactivation of oxidant scavenging enzymes that lead to oxidative stress. The observed pattern of oxidant-inducible msrA expression in Xanthomonas differs from previous reports of other bacteria, in which induction of msrA expression has been observed in response to a shift in pH (26), exposure to phenolic compounds (22), and treatment with cell wall-active antibiotics but not to oxidative stress (19). Since X. campestris pv.…”

Expression of glutathioneS-transferase and peptide methionine sulphoxide reductase inOchrobactrum anthropiis correlated to the production of reactive oxygen species caused by aromatic substrates

“…In Fig. A direct correlation between the expression and activity, respectively, of GST and MsrA was also shown for bacteria in the presence of toxic concentrations of aromatic substrates for O. anthropi [11,12]. A dose-dependent relation between the ROS production and the expression levels of both proteins can be observed.…”

“…In O. anthropi, a bacterium able to degrade xenobiotic pollutants such as atrazine, phenol and 4-chlorophenol, the presence of toxic concentrations of these xenobiotic substrates led to an increase in the expression of a monocistronically and constitutively transcribed GST [11]. In O. anthropi, the gst gene was located upstream of a gene coding for a peptide methionine sulphoxide reductase (MsrA) [12]. MsrA is a ubiquitous enzyme which is found in a wide variety of organisms and animal tissues [13].…”

Oxidative stress and heat-shock responses in Desulfovibrio vulgaris by genome-wide transcriptomic analysis

“…The induction of the msr genes in response to oxidative stress was predictable as previous studies had shown that E. coli and S. cerevisiae strains with disrupted msr genes are more sensitive to oxidative stress (Moskovitz et al 1995(Moskovitz et al , 1997(Moskovitz et al , 1998. Recent studies showed that expression of msr genes was also responsive to several chemical stress treatments, including heavy metal stress in E. faecalis (Laplace et al 2000) and phenol and 4-chlorophenol stress in Ochrobactrum anthropi (Tamburro et al 2001). Another study showed that a set of highly conserved methionine residues in Hsp21, a chloroplast-localized small heat shock protein, can become sulfoxidized and thereafter reduced back to methionines by Msr in Arabidopsis thaliana (Gustavsson et al 2002).…”