The peroxiredoxin Tpx1 is essential as a H2O2 scavenger during aerobic growth in fission yeast.
Peroxiredoxins are known to interact with hydrogen peroxide (H(2)O(2)) and to participate in oxidant scavenging, redox signal transduction, and heat-shock responses. The two-cysteine peroxiredoxin Tpx1 of Schizosaccharomyces pombe has been characterized as the H(2)O(2) sensor that transduces the redox signal to the transcription factor Pap1. Here, we show that Tpx1 is essential for aerobic, but not anaerobic, growth. We demonstrate that Tpx1 has an exquisite sensitivity for its substrate, which explains its participation in maintaining low steady-state levels of H(2)O(2). We also show in vitro and in vivo that inactivation of Tpx1 by oxidation of its catalytic cysteine to a sulfinic acid is always preceded by a sulfinic acid form in a covalently linked dimer, which may be important for understanding the kinetics of Tpx1 inactivation. Furthermore, we provide evidence that a strain expressing Tpx1.C169S, lacking the resolving cysteine, can sustain aerobic growth, and we show that small reductants can modulate the activity of the mutant protein in vitro, probably by supplying a thiol group to substitute for cysteine 169.
Jara
Mónica
M
Departament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, E-08003 Barcelona, Spain.
Vivancos
Ana P
AP
Calvo
Isabel A
IA
Moldón
Alberto
A
Sansó
Miriam
M
Hidalgo
Elena
E
D000332:Aerobiosis / Q000502:physiology*; D000818:Animals; D003545:Cysteine / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism*; D016877:Oxidants / Q000378:metabolism*; D010084:Oxidation-Reduction; D000073718:Pancreatitis-Associated Proteins; D010544:Peroxidases / Q000235:genetics / Q000378:metabolism*; D054464:Peroxiredoxins; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000502:physiology*; D029702:Schizosaccharomyces pombe Proteins; D013379:Substrate Specificity; D013441:Sulfinic Acids / Q000378:metabolism
D016877:Oxidants; D000073718:Pancreatitis-Associated Proteins; D020033:Protein Isoforms; C000612141:REG3A protein, human; D029702:Schizosaccharomyces pombe Proteins; D013441:Sulfinic Acids; C515985:Tpx1 protein, S pombe; D006861:Hydrogen Peroxide; D010544:Peroxidases; D054464:Peroxiredoxins; D003545:Cysteine