oxydo-reducteur systems and the thiol-dependent antioxidant equipment in melampsora benjamin selles,...

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 peroxidasesperoxiredoxins & glutathione peroxidasessome specific glutathione-S-transferases

• Methionine sulfoxide reductases : indirect massive detoxification through MetSO reduction

ROOH + ROH + H2ORSH2 RS2+

MetSO + Met + H2ORSH2 RS2+

H2O2

2H2O + O2H2O2 + H2O2

• Superoxide dismutases 02.- H2O2 + 0202

.-+2 H+ + 2 e-

SOD, catalases, thiol-peroxidases, methionine sulfoxide reductases

Thiol peroxidase phylogenetic tree

MlpPrxQ

PrxQ

2-cysPrx

1-cysPrxMlp1-cys

PrxII

MlpPrxII

Gpx MlpGpx

Methionine sulfoxide reductases phylogenetic tree

MlpMsrBMlpMsrA

Glutathione-S-transferases

►Complex and widespread superfamilyComplex and widespread superfamily►Share thioredoxin foldShare thioredoxin fold►Prominent roles : Prominent roles :

phase II detoxication enzymesphase II detoxication enzymes Glutathione peroxidaseGlutathione peroxidase Nitrogen metabolism…Nitrogen metabolism…

►Poorly characterized in fungiPoorly characterized in fungi

Organism Physiology GST coding sequences

Ascomycetes

Candida albicans Human pathogen 10

Saccharomyces cerevisiae Baker yeast 11

Yarrowia lipolytica Hydrocarbons using yeast 12

Neurospora crassa Red bread mold 12

Magnaporthe grisea Plant pathogen 13

Aspergillus nidulans Human pathogen 17

Botrytis cinerea Plant pathogen 21

Sclerotinia sclerotiorum Plant pathogen 23

Aspergillus fumigatus Saprophyte and pathogen 23

Trichoderma reesei Cellulolytic 37

Basidiomycetes Melampsora laricis-populina Plant pathogen 9

Cryptococcus neoformans Plant and animal pathogen 9

Ustilago maydis Plant pathogen 11

Laccaria bicolor Ectomycorrhizal 21

Phanerochaete chrysosporium White rot fungus 28

Coprinus cinereus Saprophytic fungus 28

Postia placenta Brown rot fungus 36

Zygomycetes

Rhizopus oryzae Human pathogen 13

Phycomyces blakesleeanus Light response fungus 16

A. nidulans (AN10695)L. bicolor (LACBI1231002)

C. cinereus (CC1G06890.1) P. chysosporium (EU791893) P. chysosporium (Phchr7168) P. chysosporium (Phchr7169) N. crassa (NCU00549)

C. cinereus (CC1G10591.1) L. bicolor (LACBI1299218)

P. chrysosporium (Phchr3911) P. chrysosporium (Phchr6880)

P. chrysosporium (Phchr6881) L. bicolor (LACBI1330814) N. crassa (NCU01347.3) L. bicolor (LACBI1189790)

A. nidulans (AN3299) P. chrysosporium (Phchr6766) C. cinereus (CC1G12668.1) L. bicolor (LACBI1300632) P. chrysosporium (Phchr6683)

A. nidulans (AN4905) N. crassa (NCU04109)

A. nidulans (AN9299) C. cinereus (CC1G00247.1)


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)


S. cerevisiae (YNL229C) C. cinereus (CC1G10742.1) C. cinereus (CC1G10750.1)

L. bicolor (LACBI1184665) L. bicolor (LACBI1296671) L. bicolor (LACBI1298606) C. cinereus (CC1G11890.1) C. cinereus (CC1G12718.1) P. chrysosporium (Phchr7971)

L. bicolor (LACBI1240633) L. bicolor (LACBI1314362)

S. cerevisiae (YLL060C) A. nidulans (AN10038) A. nidulans (AN1595)


S. cerevisiae (YKL081W) S. cerevisiae (YPL048W) C. cinereus (CC1G01604.1) L. bicolor (LACBI1248920) P. chrysosporium (Phchr39727) S. cerevisiae (YGR201C)

A. nidulans (AN9304) N. crassa (NCU10521.3) A. nidulans (AN0629)

N. crassa (NCU05706) C. cinereus (CC1G13373.1) L. bicolor (LACBI1319137) S. cerevisiae (YIR038C)

A. nidulans (AN10273) N. crassa (NCU04368) C. cinereus (CC1G04116.1)

C. cinereus (CC1G11081.1) L. bicolor (LACBI1179508) 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 (CC1G00881.1) C. cinereus (CC1G01486.1) C. cinereus (CC1G04959.1) C. cinereus (CC1G12612.1)

C. cinereus (CC1G05384.1) L. bicolor (LACBI1238438) L. bicolor (LACBI1295178) P. chrysosporium (Phchr5119) P. chrysosporium (Phchr5118)

P. chrysosporium (Phchr5122) P. chrysosporium (Phchr5300) C. cinereus (CC1G02842.1) C. cinereus (CC1G05417.1)

C. cinereus (CC1G07900.1) C. cinereus (CC1G07903.1) C. cinereus (CC1G07902.1)

C. cinereus (CC1G07927.1) C. cinereus (CC1G07928.1)

C. cinereus (CC1G02394.1) C. cinereus (CC1G07746.1) L. bicolor (LACBI1318115)

P. chrysosporium (Phchr7058) L. bicolor (LACBI1311800) A. nidulans (AN6612) N. crassa (NCU04150) S. cerevisiae (YAL025C)

C. cinereus (CC1G05311.1) L. bicolor (LACBI1146679) P. chrysosporium (Phchr137531)

N. crassa (NCU06035.3) S. cerevisiae (YAL003W) L. bicolor (LACBI1328850)

P. chrysosporium (Phchr1288) L. bicolor (LACBI1245857)

0.1

Cluster3/Omega-like

Ure2p

Cluster

2

GTT2

EF1B

Cluster1/GTT1

Omega

Etherases

MAK16

Figure 1

4

1

1

2

1

































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

1

1

2

1

































0.1

Cluster3/Omega-like

Ure2p

Cluster

2

GTT2

EF1B

Cluster1/GTT1

Omega

Etherases

MAK16

Figure 1

4

1

1: Elongation factor

2

1

































0.1

Cluster3/Omega-like

Ure2p

Cluster

2

GTT2

EF1B

Cluster1/GTT1

Omega

Etherases

MAK16

Figure 1

4

1

1

2

1: Glutathione peroxidase

































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

































0.1

Cluster3/Omega-like

Ure2p

Cluster

2

GTT2

EF1B

Cluster1/GTT1

Omega

Etherases

MAK16

Figure 1

4

1

1

2 ?

1

Oxidation

ReductionSH

SH

SH

Disulfide bond formation, reduction, isomerisation

GRX

TRX, GRX

SOXPDI

Regulation of protein folding, activity

1. Spontaneous (with oxygen)2. Catalyzed by SOX3. Concomitant to a reducing activity

S

S

SH

SH

S

S

SSG

SH

SH

CxxC/S redox centers

PRX

GPX

ROS

Grx

GR

GSH NTR

Trx

NADPH

Msr

Trx

S-SGROS

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 domainsCGFS

Fe/S cluster formation

CGFS

DithiolCPYC

CPYS

TRX TRX

Erv1, TRX and CytC interaction in IMS; H2O production

Oxidation and isomerization by QSOX (Erv2 for fungi) and

PDI on unfolded proteins

Erv1 and TRX interaction in IMS; H2O2 production

Disulfide bond formation by sulfhydryl oxidasesand 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 biogenesisProtein folding

Cytosolic Fe-S protein maturation

QSOXQSOX1 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

Firstisolated from liver in 1963 Intracellular localisation is:

ER

Folding of proteins by isomerization of disulfide

bonds

Protein folding

CXXC CXXCN C

Domaine TRX 1 Domaine TRX 2

CXXC CXXCN 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_gw1.18.104.1)

Mellp (e_gw1.25.122.1)

Ascomycete

Ascomycete

Ascomycete

BasidiomyceteBasidiomycete

Basidiomycete

BasidiomyceteAsc

PDI class 2

PDI class 1

PDI class 5

EuGene.00020304

estExt_fgenesh2_pg.C_450075

estExt_fgenesh2_pg.C_240082

EuGene.00190108

02.- H2O2SOD OH.

Fe2+, Cu 2+

Fenton reaction

H2O + 1/2O2

Cat

NO.

OONO-

H2OSensingby TF

Trx, TpxNO2

Tpx, Grx, GST

RSNORSH

Tpx

Trx?

Protein thiol oxidation

ROS/RNS sensing and degradation

R(SH)2 RSSR, RSSR’, R(S)2RSSG

RSOHGSH

Grx, Srx? Trx, Grx

Grx? Trx, Grx

RSO2H RSO3H

Srx

PL-OOH

Tpx

PL

Conclusions

sulfenic

sulfinic sulfonic

Heme- thiolate Haloperoxidases

Hofrichter and Ulrich, 2006

2CPO 4 GSGIG.[1].PYDNNTLPYVAPG.[2].DSRAPCPALNALANHGYIPHDGRAIS.[4].QNAFLNHMGIANSVIELALT 74 gi 2498968 21 GSAAA ELDFEQWHPAGLG DLRCGCPAMNSLANHGFINHNGSNIT VNEVIPLMQEVFHLSEELAT 84 2CPO 75 .[1].AFVVCEYVTGSDCGDSLVNLTLLAEPHAFEHDHSFSRKD YKQGVANSND FIDNRN FDAET 135 gi 2498968 85 IVTGLAVLSADDPASGIFNLDMLNRHNIFEHDASLTRKD.[1].YLGGDGHTID.[2].TLDEFL.[2].FDGKE 149 2CPO 136 FQTSLDVVAGKTH FDYADMN EIRLQRESLSNELDFPG WF TESKPIQN VESGFIFA 190 gi 2498968 150 WIDLNDAAAARYA.[2].LDSREKN.[1].SFLYQDQQLITSYGETI.[1].YF.[1].TMVDPRSN.[1].TSAEFVRI 210 2CPO 191 LVSDFNLP.[1].NDENPLVRIDW WKYW.[1].TNESF.[4].GWH.[1].PSPAREIEFVTSASSAVLAASVTS 252 gi 2498968 211 LFTEERLP VRKGGSAREKR.[2].GSRW PAMSF SWR CAPQRSSLACRSTSVRLQSRPLTR 267 2CPO 253 TP SSLPSG.[2].GPGAEAVPLSFASTMTPFLLAT.[1].APY YAQDPTLGP 297 gi 2498968 268 CH.[1].SGLPSG LPRTTRVSVRGISLSLSGGLRR RPF.[1].FVLDRHLSF 311

Pfam 1328: Peroxidase_2

Peroxidase, family 2. The peroxidases in this family do not have similarity to other peroxidases.

BLAST withESTEXT_FGENESH1_PG.C_240160

Ajellomyces capsulatus NAm1 0Ashbya gossypii ATCC 10895 0Aspergillus clavatus NRRL 1 0Aspergillus nidulans 4(5)Aspergillus flavus 0Aspergillus fumigatus 1(2)Aspergillus niger 2Aspergillus oryzae 2Botryotinia fuckeliana B05.10 7Candida albicans 0Candida tropicalis 0Chaetomium globosum 3Clavispora lusitaniae ATCC 42720 0Coccidioides immitis RS 0Cryptococcus neoformans var. neoformans JEC21 0Coprinopsis cinerea okayama7#130 5Debaryomyces hansenii CBS767 0Fusarium oxysporum 0Gibberella moniliformis 0Laccaria bicolor 1(2)Magnaporthe grisea 4(5)Malassezia globosa CBS 7966 0Melampsora 16Mycosphaerella graminicola 15(16)Nectria haematococca mpVI 0(3)Neosartorya fischeri NRRL 181 0Neurospora crassa OR74A 1Phaeosphaeria nodorum SN15 10(11)Phanerochaete chrysosporium RP-78 1(4)Phycomyces blakesleeanus 0Pichia stipitis CBS 6054 0Postia placenta 0Puccinia graminis f. sp. tritici CRL 75-36-700-3 0Rhizopus oryzae RA 99-880 0Saccharomyces cerevisiae 0Schizosaccharomyces pombe 972h- 0Sclerotinia sclerotiorum 1980 4Sporobolomyces roseus IAM 13481 0Trichoderma reesei QM6a 1Uncinocarpus reesii 1704 0Ustilago maydis 521 0Yarrowia lipolytica CLIB122 0

oxydo-reducteur systems and the thiol-dependent antioxidant equipment in melampsora benjamin selles,...

Documents

chrysosporium phchr126388

chrysosporium phchr128511

chrysosporium phchr6881

chrysosporium phchr2268

chrysosporium phchr140156

chrysosporium phchr6880

chrysosporium phchr140271

chrysosporium phchr137250