C-terminal truncation of the peroxiredoxin Tpx1 decreases its sensitivity for hydrogen peroxide without compromising its role in signal transduction.
Peroxiredoxins (Prxs) participate in hydrogen peroxide (H2O2) scavenging. Eukaryotic Prxs suffer H2O2-dependent inactivation, due to the oxidation of its catalytic cysteine to sulfinic acid, a modification which can be enzymatically reversed. This substrate-mediated reversible inactivation has been suggested to allow eukaryotic Prxs to act as floodgates, permitting high levels of H2O2 to trigger signal transduction. To test this hypothesis, we used the fission yeast Prx Tpx1, which acts as a H2O2 scavenger during aerobic metabolism and also participates in peroxide-induced signal transduction pathways. High concentrations of peroxide reversibly inactivate Tpx1.Here, we describe the characterization of a Tpx1 derivative, which lacks a carboxy-terminal extension present only in eukaryotic Prxs. This mutant protein is not inactivated by high doses of H2O2. Exclusive expression of this truncated version of Tpx1 is deleterious for aerobic growth, but H2O2-dependent signal transduction is not impaired in this strain. Instead, the ability of Tpx1.DeltaCTD to detect and detoxify peroxides is impaired. Our results indicate that inactivation of Tpx1 by excess peroxides is not required for H2O2 signaling towards the Sty1 pathway, as expected from the floodgate model, and that the carboxy-terminal extension of Tpx1 concomitantly improves H2O2 scavenging and increases susceptibility to inactivation.
Jara
Mónica
M
Oxidative Stress and Cell Cycle Group, Universitat Pompeu Fabra, C/Dr. Aiguader 88, E-08003 Barcelona, Spain.
Vivancos
Ana P
AP
Hidalgo
Elena
E
D000332:Aerobiosis; D050976:Basic-Leucine Zipper Transcription Factors / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006861:Hydrogen Peroxide / Q000378:metabolism* / Q000494:pharmacology; D007700:Kinetics; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D018384:Oxidative Stress; D054464:Peroxiredoxins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion; D015398:Signal Transduction; D013441:Sulfinic Acids / Q000378:metabolism
D050976:Basic-Leucine Zipper Transcription Factors; C105096:Pap1 protein, S pombe; D011994:Recombinant Proteins; D029702:Schizosaccharomyces pombe Proteins; D013441:Sulfinic Acids; C515985:Tpx1 protein, S pombe; D006861:Hydrogen Peroxide; D054464:Peroxiredoxins; D020928:Mitogen-Activated Protein Kinases; C095409:sty1 protein, S pombe