Oxydo-reducteur systems and the thiol-dependent antioxidant equipment in Melampsora Benjamin Selles, Nicolas Rouhier, Eric Gelhaye.

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Oxydo-reducteur systems and the thiol-dependent antioxidant equipment in Melampsora Benjamin Selles, Nicolas Rouhier, Eric Gelhaye

Oxygen is essential, but highly toxic Formation of reactive oxygen species (ROS) due to electron transport chains and oxidase functioning Release of ROS by plants as defense mechanism against pathogens Diffusion within cells : importance to have an antioxidant equipment in all sub-cellular compartments Oxydo-reducteur systems and the thiol- dependent antioxidant equipment in Melampsora

ROS formation

Superoxide and peroxide scavenging Catalases : haem peroxidase (peroxisomes) Thiol peroxidases : haem free thiol dependent peroxidases peroxiredoxins & glutathione peroxidases some specific glutathione-S-transferases Methionine sulfoxide reductases : indirect massive detoxification through MetSO reduction ROOH +ROH+ H2OH2O RSH 2 RS 2 + MetSO +Met+ H2OH2O RSH 2 RS 2 + H2O2H2O2 2H 2 O + O 2 H 2 O 2 + H 2 O 2 Superoxide dismutases H 2 O H e -

SOD, catalases, thiol-peroxidases, methionine sulfoxide reductases

Thiol peroxidase phylogenetic tree MlpPrxQ PrxQ 2-cysPrx 1-cysPrx Mlp1-cys PrxII MlpPrxII Gpx MlpGpx

Methionine sulfoxide reductases phylogenetic tree MlpMsrB MlpMsrA

Glutathione-S-transferases ► Complex and widespread superfamily ► Share thioredoxin fold ► Prominent roles :  phase II detoxication enzymes  Glutathione peroxidase  Nitrogen metabolism… ► Poorly characterized in fungi

A. nidulans (AN10695) L. bicolor (LACBI ) C. cinereus (CC1G ) P. chysosporium (EU791893) P. chysosporium (Phchr7168) P. chysosporium (Phchr7169) N. crassa (NCU00549) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr3911) P. chrysosporium (Phchr6880) P. chrysosporium (Phchr6881) L. bicolor (LACBI ) N. crassa (NCU ) L. bicolor (LACBI ) A. nidulans (AN3299) P. chrysosporium (Phchr6766) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr6683) A. nidulans (AN4905) N. crassa (NCU04109) A. nidulans (AN9299) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr503) A. nidulans (AN3255) N. crassa (NCU05780) P. chrysosporium (Phchr140156) P. chrysosporium (Phchr137250) P. chrysosporium (Phchr2269) P. chrysosporium (Phchr2266) P. chrysosporium (Phchr140259) P. chrysosporium (Phchr140271) P. chrysosporium (Phchr2268) P. chrysosporium (Phchr128511) S. cerevisiae (YNL229C) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) L. bicolor (LACBI ) L. bicolor (LACBI ) C. cinereus (CC1G ) C. cinereus (CC1G ) P. chrysosporium (Phchr7971) L. bicolor (LACBI ) L. bicolor (LACBI ) S. cerevisiae (YLL060C) A. nidulans (AN10038) A. nidulans (AN1595) A. nidulans (AN6563) N. crassa (NCU03826) S. cerevisiae (YKL081W) S. cerevisiae (YPL048W) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr39727) S. cerevisiae (YGR201C) A. nidulans (AN9304) N. crassa (NCU ) A. nidulans (AN0629) N. crassa (NCU05706) C. cinereus (CC1G ) L. bicolor (LACBI ) S. cerevisiae (YIR038C) A. nidulans (AN10273) N. crassa (NCU04368) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (EU791894) P. chrysosporium (Phchr126388) S. cerevisiae (YGR154C) S. cerevisiae (YKR076W) S. cerevisiae (YMR251W) A. nidulans (AN5831) N. crassa (NCU09570) A. nidulans (AN10379) A. nidulans (AN2592) A. nidulans (AN2948) N. crassa (NCU02888) A. nidulans (AN8942) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) L. bicolor (LACBI ) P. chrysosporium (Phchr5119) P. chrysosporium (Phchr5118) P. chrysosporium (Phchr5122) P. chrysosporium (Phchr5300) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr7058) L. bicolor (LACBI ) A. nidulans (AN6612) N. crassa (NCU04150) S. cerevisiae (YAL025C) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr137531) N. crassa (NCU ) S. cerevisiae (YAL003W) L. bicolor (LACBI ) P. chrysosporium (Phchr1288) L. bicolor (LACBI ) 0.1 Cluster3/Omega-like Ure2p Cluster 2 GTT2 EF1B Cluster1/GTT1 Omega Etherases MAK16 Figure

A. nidulans (AN10695) L. bicolor (LACBI ) C. cinereus (CC1G ) P. chysosporium (EU791893) P. chysosporium (Phchr7168) P. chysosporium (Phchr7169) N. crassa (NCU00549) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr3911) P. chrysosporium (Phchr6880) P. chrysosporium (Phchr6881) L. bicolor (LACBI ) N. crassa (NCU ) L. bicolor (LACBI ) A. nidulans (AN3299) P. chrysosporium (Phchr6766) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr6683) A. nidulans (AN4905) N. crassa (NCU04109) A. nidulans (AN9299) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr503) A. nidulans (AN3255) N. crassa (NCU05780) P. chrysosporium (Phchr140156) P. chrysosporium (Phchr137250) P. chrysosporium (Phchr2269) P. chrysosporium (Phchr2266) P. chrysosporium (Phchr140259) P. chrysosporium (Phchr140271) P. chrysosporium (Phchr2268) P. chrysosporium (Phchr128511) S. cerevisiae (YNL229C) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) L. bicolor (LACBI ) L. bicolor (LACBI ) C. cinereus (CC1G ) C. cinereus (CC1G ) P. chrysosporium (Phchr7971) L. bicolor (LACBI ) L. bicolor (LACBI ) S. cerevisiae (YLL060C) A. nidulans (AN10038) A. nidulans (AN1595) A. nidulans (AN6563) N. crassa (NCU03826) S. cerevisiae (YKL081W) S. cerevisiae (YPL048W) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr39727) S. cerevisiae (YGR201C) A. nidulans (AN9304) N. crassa (NCU ) A. nidulans (AN0629) N. crassa (NCU05706) C. cinereus (CC1G ) L. bicolor (LACBI ) S. cerevisiae (YIR038C) A. nidulans (AN10273) N. crassa (NCU04368) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (EU791894) P. chrysosporium (Phchr126388) S. cerevisiae (YGR154C) S. cerevisiae (YKR076W) S. cerevisiae (YMR251W) A. nidulans (AN5831) N. crassa (NCU09570) A. nidulans (AN10379) A. nidulans (AN2592) A. nidulans (AN2948) N. crassa (NCU02888) A. nidulans (AN8942) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) L. bicolor (LACBI ) P. chrysosporium (Phchr5119) P. chrysosporium (Phchr5118) P. chrysosporium (Phchr5122) P. chrysosporium (Phchr5300) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr7058) L. bicolor (LACBI ) A. nidulans (AN6612) N. crassa (NCU04150) S. cerevisiae (YAL025C) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr137531) N. crassa (NCU ) S. cerevisiae (YAL003W) L. bicolor (LACBI ) P. chrysosporium (Phchr1288) L. bicolor (LACBI ) 0.1 Cluster3/Omega-like Ure2p Cluster 2 GTT2 EF1B Cluster1/GTT1 Omega Etherases MAK16 Figure 1 4 : oxidative stress, Glutathione peroxidase, nitrogen metabolism regulation

A. nidulans (AN10695) L. bicolor (LACBI ) C. cinereus (CC1G ) P. chysosporium (EU791893) P. chysosporium (Phchr7168) P. chysosporium (Phchr7169) N. crassa (NCU00549) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr3911) P. chrysosporium (Phchr6880) P. chrysosporium (Phchr6881) L. bicolor (LACBI ) N. crassa (NCU ) L. bicolor (LACBI ) A. nidulans (AN3299) P. chrysosporium (Phchr6766) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr6683) A. nidulans (AN4905) N. crassa (NCU04109) A. nidulans (AN9299) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr503) A. nidulans (AN3255) N. crassa (NCU05780) P. chrysosporium (Phchr140156) P. chrysosporium (Phchr137250) P. chrysosporium (Phchr2269) P. chrysosporium (Phchr2266) P. chrysosporium (Phchr140259) P. chrysosporium (Phchr140271) P. chrysosporium (Phchr2268) P. chrysosporium (Phchr128511) S. cerevisiae (YNL229C) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) L. bicolor (LACBI ) L. bicolor (LACBI ) C. cinereus (CC1G ) C. cinereus (CC1G ) P. chrysosporium (Phchr7971) L. bicolor (LACBI ) L. bicolor (LACBI ) S. cerevisiae (YLL060C) A. nidulans (AN10038) A. nidulans (AN1595) A. nidulans (AN6563) N. crassa (NCU03826) S. cerevisiae (YKL081W) S. cerevisiae (YPL048W) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr39727) S. cerevisiae (YGR201C) A. nidulans (AN9304) N. crassa (NCU ) A. nidulans (AN0629) N. crassa (NCU05706) C. cinereus (CC1G ) L. bicolor (LACBI ) S. cerevisiae (YIR038C) A. nidulans (AN10273) N. crassa (NCU04368) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (EU791894) P. chrysosporium (Phchr126388) S. cerevisiae (YGR154C) S. cerevisiae (YKR076W) S. cerevisiae (YMR251W) A. nidulans (AN5831) N. crassa (NCU09570) A. nidulans (AN10379) A. nidulans (AN2592) A. nidulans (AN2948) N. crassa (NCU02888) A. nidulans (AN8942) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) L. bicolor (LACBI ) P. chrysosporium (Phchr5119) P. chrysosporium (Phchr5118) P. chrysosporium (Phchr5122) P. chrysosporium (Phchr5300) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr7058) L. bicolor (LACBI ) A. nidulans (AN6612) N. crassa (NCU04150) S. cerevisiae (YAL025C) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr137531) N. crassa (NCU ) S. cerevisiae (YAL003W) L. bicolor (LACBI ) P. chrysosporium (Phchr1288) L. bicolor (LACBI ) 0.1 Cluster3/Omega-like Ure2p Cluster 2 GTT2 EF1B Cluster1/GTT1 Omega Etherases MAK16 Figure : Elongation factor 2 1

A. nidulans (AN10695) L. bicolor (LACBI ) C. cinereus (CC1G ) P. chysosporium (EU791893) P. chysosporium (Phchr7168) P. chysosporium (Phchr7169) N. crassa (NCU00549) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr3911) P. chrysosporium (Phchr6880) P. chrysosporium (Phchr6881) L. bicolor (LACBI ) N. crassa (NCU ) L. bicolor (LACBI ) A. nidulans (AN3299) P. chrysosporium (Phchr6766) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr6683) A. nidulans (AN4905) N. crassa (NCU04109) A. nidulans (AN9299) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr503) A. nidulans (AN3255) N. crassa (NCU05780) P. chrysosporium (Phchr140156) P. chrysosporium (Phchr137250) P. chrysosporium (Phchr2269) P. chrysosporium (Phchr2266) P. chrysosporium (Phchr140259) P. chrysosporium (Phchr140271) P. chrysosporium (Phchr2268) P. chrysosporium (Phchr128511) S. cerevisiae (YNL229C) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) L. bicolor (LACBI ) L. bicolor (LACBI ) C. cinereus (CC1G ) C. cinereus (CC1G ) P. chrysosporium (Phchr7971) L. bicolor (LACBI ) L. bicolor (LACBI ) S. cerevisiae (YLL060C) A. nidulans (AN10038) A. nidulans (AN1595) A. nidulans (AN6563) N. crassa (NCU03826) S. cerevisiae (YKL081W) S. cerevisiae (YPL048W) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr39727) S. cerevisiae (YGR201C) A. nidulans (AN9304) N. crassa (NCU ) A. nidulans (AN0629) N. crassa (NCU05706) C. cinereus (CC1G ) L. bicolor (LACBI ) S. cerevisiae (YIR038C) A. nidulans (AN10273) N. crassa (NCU04368) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (EU791894) P. chrysosporium (Phchr126388) S. cerevisiae (YGR154C) S. cerevisiae (YKR076W) S. cerevisiae (YMR251W) A. nidulans (AN5831) N. crassa (NCU09570) A. nidulans (AN10379) A. nidulans (AN2592) A. nidulans (AN2948) N. crassa (NCU02888) A. nidulans (AN8942) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) L. bicolor (LACBI ) P. chrysosporium (Phchr5119) P. chrysosporium (Phchr5118) P. chrysosporium (Phchr5122) P. chrysosporium (Phchr5300) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr7058) L. bicolor (LACBI ) A. nidulans (AN6612) N. crassa (NCU04150) S. cerevisiae (YAL025C) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr137531) N. crassa (NCU ) S. cerevisiae (YAL003W) L. bicolor (LACBI ) P. chrysosporium (Phchr1288) L. bicolor (LACBI ) 0.1 Cluster3/Omega-like Ure2p Cluster 2 GTT2 EF1B Cluster1/GTT1 Omega Etherases MAK16 Figure : Glutathione peroxidase

A. nidulans (AN10695) L. bicolor (LACBI ) C. cinereus (CC1G ) P. chysosporium (EU791893) P. chysosporium (Phchr7168) P. chysosporium (Phchr7169) N. crassa (NCU00549) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr3911) P. chrysosporium (Phchr6880) P. chrysosporium (Phchr6881) L. bicolor (LACBI ) N. crassa (NCU ) L. bicolor (LACBI ) A. nidulans (AN3299) P. chrysosporium (Phchr6766) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr6683) A. nidulans (AN4905) N. crassa (NCU04109) A. nidulans (AN9299) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr503) A. nidulans (AN3255) N. crassa (NCU05780) P. chrysosporium (Phchr140156) P. chrysosporium (Phchr137250) P. chrysosporium (Phchr2269) P. chrysosporium (Phchr2266) P. chrysosporium (Phchr140259) P. chrysosporium (Phchr140271) P. chrysosporium (Phchr2268) P. chrysosporium (Phchr128511) S. cerevisiae (YNL229C) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) L. bicolor (LACBI ) L. bicolor (LACBI ) C. cinereus (CC1G ) C. cinereus (CC1G ) P. chrysosporium (Phchr7971) L. bicolor (LACBI ) L. bicolor (LACBI ) S. cerevisiae (YLL060C) A. nidulans (AN10038) A. nidulans (AN1595) A. nidulans (AN6563) N. crassa (NCU03826) S. cerevisiae (YKL081W) S. cerevisiae (YPL048W) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr39727) S. cerevisiae (YGR201C) A. nidulans (AN9304) N. crassa (NCU ) A. nidulans (AN0629) N. crassa (NCU05706) C. cinereus (CC1G ) L. bicolor (LACBI ) S. cerevisiae (YIR038C) A. nidulans (AN10273) N. crassa (NCU04368) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (EU791894) P. chrysosporium (Phchr126388) S. cerevisiae (YGR154C) S. cerevisiae (YKR076W) S. cerevisiae (YMR251W) A. nidulans (AN5831) N. crassa (NCU09570) A. nidulans (AN10379) A. nidulans (AN2592) A. nidulans (AN2948) N. crassa (NCU02888) A. nidulans (AN8942) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) L. bicolor (LACBI ) P. chrysosporium (Phchr5119) P. chrysosporium (Phchr5118) P. chrysosporium (Phchr5122) P. chrysosporium (Phchr5300) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr7058) L. bicolor (LACBI ) A. nidulans (AN6612) N. crassa (NCU04150) S. cerevisiae (YAL025C) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr137531) N. crassa (NCU ) S. cerevisiae (YAL003W) L. bicolor (LACBI ) P. chrysosporium (Phchr1288) L. bicolor (LACBI ) 0.1 Cluster3/Omega-like Ure2p Cluster 2 GTT2 EF1B Cluster1/GTT1 Omega Etherases MAK16 Figure 1 4 1: Dihydroascorbate reductase, thiol transferase 1 2 1

A. nidulans (AN10695) L. bicolor (LACBI ) C. cinereus (CC1G ) P. chysosporium (EU791893) P. chysosporium (Phchr7168) P. chysosporium (Phchr7169) N. crassa (NCU00549) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr3911) P. chrysosporium (Phchr6880) P. chrysosporium (Phchr6881) L. bicolor (LACBI ) N. crassa (NCU ) L. bicolor (LACBI ) A. nidulans (AN3299) P. chrysosporium (Phchr6766) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr6683) A. nidulans (AN4905) N. crassa (NCU04109) A. nidulans (AN9299) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr503) A. nidulans (AN3255) N. crassa (NCU05780) P. chrysosporium (Phchr140156) P. chrysosporium (Phchr137250) P. chrysosporium (Phchr2269) P. chrysosporium (Phchr2266) P. chrysosporium (Phchr140259) P. chrysosporium (Phchr140271) P. chrysosporium (Phchr2268) P. chrysosporium (Phchr128511) S. cerevisiae (YNL229C) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) L. bicolor (LACBI ) L. bicolor (LACBI ) C. cinereus (CC1G ) C. cinereus (CC1G ) P. chrysosporium (Phchr7971) L. bicolor (LACBI ) L. bicolor (LACBI ) S. cerevisiae (YLL060C) A. nidulans (AN10038) A. nidulans (AN1595) A. nidulans (AN6563) N. crassa (NCU03826) S. cerevisiae (YKL081W) S. cerevisiae (YPL048W) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr39727) S. cerevisiae (YGR201C) A. nidulans (AN9304) N. crassa (NCU ) A. nidulans (AN0629) N. crassa (NCU05706) C. cinereus (CC1G ) L. bicolor (LACBI ) S. cerevisiae (YIR038C) A. nidulans (AN10273) N. crassa (NCU04368) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (EU791894) P. chrysosporium (Phchr126388) S. cerevisiae (YGR154C) S. cerevisiae (YKR076W) S. cerevisiae (YMR251W) A. nidulans (AN5831) N. crassa (NCU09570) A. nidulans (AN10379) A. nidulans (AN2592) A. nidulans (AN2948) N. crassa (NCU02888) A. nidulans (AN8942) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) L. bicolor (LACBI ) P. chrysosporium (Phchr5119) P. chrysosporium (Phchr5118) P. chrysosporium (Phchr5122) P. chrysosporium (Phchr5300) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr7058) L. bicolor (LACBI ) A. nidulans (AN6612) N. crassa (NCU04150) S. cerevisiae (YAL025C) C. cinereus (CC1G ) L. bicolor (LACBI ) P. chrysosporium (Phchr137531) N. crassa (NCU ) S. cerevisiae (YAL003W) L. bicolor (LACBI ) P. chrysosporium (Phchr1288) L. bicolor (LACBI ) 0.1 Cluster3/Omega-like Ure2p Cluster 2 GTT2 EF1B Cluster1/GTT1 Omega Etherases MAK16 Figure ? 1

Oxidation Reduction SH Disulfide bond formation, reduction, isomerisation GRX TRX, GRX SOX PDI Regulation of protein folding, activity 1.Spontaneous (with oxygen) 2.Catalyzed by SOX 3.Concomitant to a reducing activity S S SH S S SSG SH CxxC/S redox centers

PRX GPX ROS Grx GR GSH NTR Trx NADPH Msr Trx S-SG ROS

Disulfide bond reduction by glutaredoxins & thioredoxins

MlpTrx4 MlpTrx1, 2, 3

MlpGrx1.1, 1.2 MlpGrx 2.2 MlpGrx 5 MlpGrx 2.1 MlpGrx 4 Trx/Grx domains CGFS Fe/S cluster formation CGFS Dithiol CPYC CPYS

TRX Erv1, TRX and CytC interaction in IMS; H 2 O production Oxidation and isomerization by QSOX (Erv2 for fungi) and PDI on unfolded proteins Erv1 and TRX interaction in IMS; H 2 O 2 production Disulfide bond formation by sulfhydryl oxidases and isomerisation by protein disulfide isomerases

SOX (ScErv1 like) and Erv2 First discovered in S cerevisiae and was essential for respiration and vegetative growth, Intracellular localisation is: IMS (SOX /scErv1 like) ER (Erv2, Fungi) Mitochondrial biogenesis Protein folding Cytosolic Fe-S protein maturation QSOX QSOX1 was initially identified from quiescent human fibroblasts and his Intracellular localisation is: ER, Golgi, secretory granules and are located at the cell surface Folding and regulation of secreted and parietal proteins PDI and PDI like First isolated from liver in 1963 Intracellular localisation is: ER Folding of proteins by isomerization of disulfide bonds Protein folding CXXC N C Domaine TRX 1Domaine TRX 2 CXXC N C Domaine TRX 1 Domaine TRX 2 CXXC N C Domaine TRX Classe 1 Classe 2 Classe 5

Comparative genomic with other sequenced fungi

Fungus scErv2 like QSOX ( HsQuiecin like) Plants Mammalian SOX (scErv1 like) Mammalian Plants Basidiomycete Ascomycete |Mellp (e_gw ) Mellp (e_gw )

Ascomycete Basidiomycete Asc PDI class 2 PDI class 1 PDI class 5 EuGene estExt_fgenesh2_pg.C_ estExt_fgenesh2_pg.C_ EuGene

H2O2H2O2 SOD OH. Fe 2+, Cu 2+ Fenton reaction H 2 O + 1/2O 2 Cat NO. OONO - H2OH2O Sensing by TF Trx, Tpx NO 2 Tpx, Grx, GST RSNORSH Tpx Trx? Protein thiol oxidation ROS/RNS sensing and degradation R(SH) 2 RSSR, RSSR’, R(S) 2 RSSG RSOH GSH Grx, Srx?Trx, Grx Grx? Trx, Grx RSO 2 H RSO 3 H Srx PL-OOH Tpx PL Conclusions sulfenic sulfinic sulfonic

H2O2H2O2 SOD OH. Fe 2+, Cu 2+ Fenton reaction H 2 O + 1/2O 2 Cat NO. OONO - H2OH2O Sensing by TF Trx, Tpx NO 2 Tpx, Grx, GST RSNORSH Tpx Trx? Protein thiol oxidation ROS/RNS sensing and degradation R(SH) 2 RSSR, RSSR’, R(S) 2 RSSG RSOH GSH Grx, Srx?Trx, Grx Grx? Trx, Grx RSO 2 H RSO 3 H Srx PL-OOH Tpx PL Conclusions sulfenic sulfinic sulfonic

Heme- thiolate Haloperoxidases Hofrichter and Ulrich, 2006

Pfam 1328: Peroxidase_2 Peroxidase, family 2. The peroxidases in this family do not have similarity to other peroxidases.

Mlp Prx 12 isoforms Peroxidase class II (Peroxibase) All predicted secreted (target P) Function ??