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Table S3: Ustilago maydis theoretical peroxisomal protein inventory PEXs Accn no. Name Function PTS1 PTS2 MTS um04191 Pex1 AAA ATPase involved recycling/undocking of Pex5 n.a. n.a. n.a. um00325 Pex2 E(3) ligase required involved in the polyubiquitination of Pex5 n.a. n.a. n.a. um06200 Pex3 Docking factor for Pex19, vesicle assembly during de novo formation n.a. n.a. n.a. um11354 Pex4 Ubiquitin-conjugating enzyme required for monoubiquitination of Pex5 n.a. n.a. n.a. um02528, Pex5 Peroxisomal matrix protein import PTS1-receptor n.a. n.a. n.a. um02421 Pex6 AAA ATPase involved recycling/undocking of Pex5 n.a. n.a. n.a. um03596 Pex7 Peroxisomal matrix protein import PTS2-receptor n.a. n.a. n.a. um02887 Pex8 Involved in the assembly of the Po matrix protein importomer n.a. n.a. n.a. um02641 Pex10 E(3) ligase for Ubc4p-dependent ubiquitination of Pex5p n.a. n.a. n.a. um04294 Pex11α/β Peroxisome elongation, membrane bending for subsequent division n.a. n.a. n.a. um00440 Pex11γ Peroxisome elongation, membrane bending for subsequent division n.a. n.a. n.a. um00297 Pex12 E(3) ligase required for Pex4-dependent monoubiquitination of Pex5p n.a. n.a. n.a. um03165 Pex13 Part of the peroxisomal membrane docking complex for Pex5 and Pex7 n.a. n.a. n.a. um02323 Pex14 Part of the peroxisomal membrane docking complex for Pex5 and Pex7 n.a. n.a. n.a. um06341 Pex16 Recruitment of Pex3 to the Po membrane, required for de novo formation n.a. n.a. n.a. um10731 Pex19 Po membrane protein import receptor, required for de novo formation n.a. n.a. n.a. um10172 Pex5/20 fusion protein Unknown, according to the Pex20 domain possible PTS2 co-receptor n.a. n.a. n.a. um01564 Pex23 Required for peroxisome assembly/ peroxisomal matrix protein import n.a. n.a. n.a. um10336 Pex24 Required for peroxisome assembly and protein import n.a. n.a. n.a. um12204 Pex26 Pex1/Pex6 anchor protein at the peroxisomal membrane n.a. n.a. n.a. um00755 Pex33 Pex14/17 fusion protein, part of the membrane docking complex for Pex5 n.a. n.a. n.a. Peroxins Other proteins involved in peroxisome biogenesis um03919 Related to FIS1 Involved in mitochondrial/peroxisomal division, possible adaptor for DNM1 n.a. n.a. n.a. um05378 Mitochondrial fission factor DNM1 Organelle fission, member of the dynamin family of GTPases n.a. n.a. n.a. um03498 Related to CAF4 - CCR4 associated factor Involved in DNM1-dependent mitochondrial/peroxisomal division n.a. n.a. n.a. um15047 Probable VPS1 Organelle fission, member of the dynamin family of GTPases n.a. n.a. n.a. um02719 Related to DnaJ-like protein1 - DJP1 Involved in peroxisomal matrix protein import in yeast n.a. n.a. n.a. um05734 Probable GTPase Rho1 Required for peroxisome proliferation in yeasts; organization of the actin n.a. n.a. n.a. um10663 Probable RHO1 GTP-binding protein cytoskeleton attached to the peroxisomal membrane n.a. n.a. n.a. Lipid metabolism – fatty acid transport/activation um01105 Peroxisomal half ABC transporter PXA2 ABC membrane transporter, fatty acid import n.a. n.a. um03945 Peroxisomal half ABC transporter PXA1 ABC membrane transporter, fatty acid import n.a. n.a. um05450 Rel. to Long-chain-fatty-acid-CoA ligase 6 Fatty acid activation by coupling to coenzyme A PKL um02583 Rel. to Very-long-chain fatty acid-CoA synth. Fatty acid activation by coupling to coenzyme A VRL um03228 FACL-like2/long-chain fatty acid coA ligase Fatty acid activation by coupling to coenzyme A PRL um11659 PCS60 - AMP-binding protein, peroxisomal High similarities to E. coli Long-chain acyl-CoA synthase AKL um02235 Rel. to Acyl-CoA synthetase Fatty acid activation by coupling to coenzyme A SKL um05869 Rel. to mito. carnitine/acyl carnitine carrier Export of fatty acids from peroxisomes? n.a. um11660 Carnitine O-acetyltransferase (CRAT) Export of fatty acids from peroxisomes SKL um11938 Peroxisomal SCP2-like protein A Fatty acid/Steroid binding/transport AKL um01850 Peroxisomal SCP2-like protein B Fatty acid/Steroid binding/transport AKL um11277 Oleate induced Po lipid transfer protein Fatty acid/Steroid binding/transport (SCP2 domain) ARL n.a. Lipid metabolism – fatty acid β-oxidation um00122 AidB-like ACAD Fatty acid β-oxidation, first step? SHL um10665 Related to ACAD with CytB5 domain Fatty acid β-oxidation, first step? SKF um01466 CytB5 domain bearing ACAD Fatty acid β-oxidation, first step? ARL um06400 ACAD11n Fatty acid β-oxidation, first step SHL um06422 ACAD11c, N-terminal domain Unknown SKL um01966 Acyl-CoA oxidase ACOX1-like A Fatty acid β-oxidation, first step SNL um02028 Acyl-CoA oxidase ACOX1-like B Fatty acid β-oxidation, first step ARL um02208 Acyl-CoA oxidase ACOX1-like C Fatty acid β-oxidation, first step AKL um04324 Acyl-CoA oxidase POX1 Fatty acid β-oxidation, first step um02097 Peroxisomal enoyl-CoA hydratase Fatty acid β-oxidation, second step SKL um11001 Peroxisomal enoyl-CoA hydratase Fatty acid β-oxidation, second step SNL um01747 MFE-2 like enoyl-CoA hydratase Fatty acid β-oxidation, second step AKL um10825 3-Hydroxyacyl-CoA dehydrogenase type 2 Fatty acid β-oxidation, third step GKL um10038 Peroxisomal multifunctional enzyme MFE2 Fatty acid β-oxidation, second & third step um01090 POT1 - acetyl-CoA C-acyltransferase Fatty acid β-oxidation, fourth step um02715 POT1-like - acetyl-CoA C-acyltransferase Fatty acid β-oxidation, fourth step um01986 Peroxisomal sterol carrier protein SCPx Fatty acid β-oxidation, fourth step Lipid metabolism – fatty acid α-oxidation um05961 α-Methylacyl-CoA racemase (AMACR) Conversion of pristanoyl-CoA and C27-bile acyl-CoA to (S)-stereoisomers um03994 2-Hydroxyacyl-CoA lyase 1 (HPCL2) Conversion 2-hydroxyphytanoyl-CoA to pristanal um11241 Fatty aldehyde dehydrogenase (AL3A2) Conversion of pristanal to pristanic acid SKL Lipid metabolism – degradation of polyunsaturated fatty acids um03158 Rel. to Δ3,Δ2-enoyl-CoA isomerase ECI2 Isomerization of 3-trans,5-cis-dienoyl-CoA to 2-trans,4-trans-dienoyl-CoA SKL um01599 Rel. to Δ3,Δ2-enoyl-CoA isomerase ECI2 Isomerization of 3-trans,5-cis-dienoyl-CoA to 2-trans,4-trans-dienoyl-CoA HKL um10273 Rel. to Δ3,Δ2-enoyl-CoA isomerase ECH1 Isomerization of 3-trans,5-cis-dienoyl-CoA to 2-trans,4-trans-dienoyl-CoA AKL um10183 Peroxisomal 2,4-dienoyl-CoA reductase Catalyzes the reduction of 2,4-dienoyl-CoA to trans-3-enoyl-CoA SRL Lipid metabolism – miscellaneous proteins um00334 Peroxisomal acyl-CoA thioester hydrolase Regulation of intraorganellar CoA-SH levels PKL um01415 Hypothetical thioester hydrolase Unknown PKL um11630 Hypothetical thioester hydrolase Unknown SKL um06465 Hypothetical thioester hydrolase Unknown SQL um11350 Rel. to Short chain acyl-CoA synthetase 3 Activation of acetate to acetyl-CoA CKL um10088 Related to Acetoacetyl-CoA synthetase Generation of acetoacetyl-CoA from 2 acetyl-CoA AKL um02182 Rel. to Dehydrogenase/reductase SDR 4 NADPH-dependent beta-ketoacyl reductase PRL um02023 Rel.to Long chain fatty alcohol oxidase FAO1 Involved in the omega-oxidation pathway of lipid degradation ARL um02862 Rel. to Ectomycorrhiza-regulated esterase Unknown SRL um10545 Rel. to Ectomycorrhiza-regulated esterase Unknown ARL Lipid metabolism – etherlipid synthesis* um11632 Rel. to DHAP acyltransferase Synthesis of acyl-DHAP from DHAP and acyl-CoA AQL um02491 Glyceraldehyde 3-phosphate dehydrogenase Conversion of glyceraldehyde 3-phosph. into 3-phospho-glyceroyl phosph. SRL um05422 Similar to FAR1 Fatty alcohol synthesis n.a. n.a. um05893 Rel. to acyl DHAP reductase Conversion of acyl dihydroxyacetone-phosphate (DHAP) into glycerolipids n.a. n.a. Lipid metabolism –synthesis of mannosylerythritol lipids (MEL) um03116 MAC1 Transfer of fatty acids to C-2 and C-3 positions of mannosylerythritol ARL um10636 MAC2 Transfer of fatty acids to C-2 and C-3 positions of mannosylerythritol AKL Amino acid metabolism um02001 Hydroxymethylglutaryl-CoA lyase Terminal step in leucine metabolism AKL um05247 Rel. to methylcrotonyl-CoA carboxylase β Leucine degradation SRM um06326 Rel. to fumarylacetoacetate hydrolase Tyrosine degradation PKL um05703 Rel. to D-amino-acid oxidase Degradation of D-amino acids SRL um11649 Rel. to D-amino acid oxidase Degradation of D-amino acids LKL um03935 Rel. to sarcosine oxidase Oxidation of L-pipecolate and L-proline ARL um03239 Probable acetolactate synthase Catalyzes the first step in valine, isoleucine biosynthesis Glycolate/glyoxylate metabolism um10682 Rel. to Hydoxyacid oxidase 1 Degradation of glycolate to glyoxylate NKL um12269 Rel. to Hydoxyacid oxidase 1 Degradation of glycolate to glyoxylate GRL um01329 Isocitrate dehydrogenase subunit 1, IDH1 Isocitrate + NAD+ = 2-oxoglutarate + CO2 + NADH NAD+ = 2-oxoglutarate + CO2 + NADH SNL SRL um01328 Isocitrate dehydrogenase subunit 2, IDH2 Isocitrate + um06111 Isocitrate dehydrogenase, IDP1/2 Isocitrate + NADP+ = 2-oxoglutarate + CO2 + NADPH GKL um15004 Probable malate synthase, glyoxysomal Acetyl-CoA + H2O + glyoxylate = (S)-malate + CoA. ARI um11161 Probable MDH1 - malate dehydrogenase (S)-malate + NAD+ = oxaloacetate + NADH ANL Oxygen metabolism and oxidation of redox equivalents um04063 Related to Epoxide hydrolase 2 C-terminal dom.: epoxide hydrolase, N-terminal domain: lipid phosphatase SKL um01852 Related to Epoxide hydrolase 1 Catalyzes the hydrolysis of arene and aliphatic epoxides dihydrodiols SKI um10746 Similar to Epoxide hydrolase Catalyzes the hydrolysis of arene and aliphatic epoxides dihydrodiols AKL um05678 Related to Glutathione S-transferase κ RX + glutathione = HX + R-S-glutathione AKL Um02610 Related to Glutathione S-transferase RX + glutathione = HX + R-S-glutathione AKL um11067 Probable Catalase 2 Degradation of hydogen peroxide: 2 H2O2 = O2 + 2 H2O ARF um10331 Rel. to peroxisomal PMP20, PRDX5 Reduction of hydrogen peroxide and alkyl hydroperoxides AKL um01947 Related to cytochrome-c peroxidase 2 ferrocytochrome c + H2O2 = 2 ferricytochrome c + 2 H2O AKL Purine and pyrimidine metabolism um00672 Probable urate oxidase (uricase) Conversion of uric acid to allantoin AKL um03264 Probable xanthine dehydrogenase/oxidase Conversion of hypoxanthine to uric acid n.a. Carbohydrate metabolism um00555 Glycerol-3-phosphate dehydrogenase (NAD) Regulation of the NAD+/NADH coenzyme equilibrium SRL um04930 Prob. Glucose 6-phosphate dehydrogenase Conversion of glucose-6-phosphate to D-glucono-1,5-lactone-6-phosphate GKM um00674 Probable Fructose-bisphosphate aldolase Condensation of DHAP + glyceraldehyde-3-phosphate to FBP GKL um11953 Related to STE23 Mammalian insulin-degrading enzyme homolog, unknown function in Po ANL um05558 Related to mammalian Lon protease Lon2p Cleavage of PTS2, degradation of misfolded polypeptides in Po SRL Regulation of intracellular polyamine concentration SRL Proteases Amine and polyamine metabolism um03524 Rel. to Po amine oxidase (Cu-containing) um05850 Rel. to Po amine oxidase Regulation of intracellular polyamine concentration NKL Oxidizes L-gulono-1,4-lactone to H2O2 and L-xylo-hexulonolactone SRL Condensation of pimeloyl-CoA and L-alanine to 8-amino-7-oxononanoate SKL Ascorbate biosynthesis um10013 Related to l-gulonolactone oxidase Biotin biosynthesis um01197 Rel.to 8-amino-7-oxononanoate synthase Possible alcohol metabolism um01872 Related to gmc type oxidoreductase Probable peroxisomal alcohol oxidase or polyamine biosynthesis SHL um05407 Rel. to Betaine aldehyde dehydrogenase (Acet)aldehyde degradation or polyamine biosynthesis AKL PKL Cleavage of Cofactors um01332 Related to CoA diphosphatase (NUDT7) Regulation of CoA and acyl-CoA levels in response to metabolic demands um06194 Rel. to NADH pyrophosphatase NUDT12 Regulation of NADH levels levels in response to metabolic demands um03979 Rel. to Nudix motif 19 (NUDT19) Regulation of CoA and acyl-CoA levels in response to metabolic demands AKL Further acyl-activating enzymes um15048 Related to 4-coumarate-CoA ligase 1 Unknown AKL um06153 Related to 4-coumarate--CoA ligase 1 Unknown ANL um04479 Related to 4-coumarate--CoA ligase Unknown AKL Miscellaneous proteins with unknown function um01189 Probable translation elongation factor eEF1β EF1B catalyzes the exchange of GDP bound to the G-protein for GTP AKI um10014 Rel. to Ribonuclease UK114 Unknown function in Po metabolism (Antonenkov et al. 2007) SNL um01286 Related to PTC5 - 2C protein phosphatase Sim. to metazoan mitochondrial pyruvate dehydrogenase phosphatase 1 AKL um11368 Probable cytochrome b5 Electron acceptor protein AKL um11567 Related to Histidine triad protein Superfamily of nucleotide-binding and -hydrolyzing enzymes AKL um00154 Related to Para-nitrobenzyl esterase Possible carboxylesterase AKL um01881 Related to Oxidoreductase SDR short-chain dehydrogenase SHL um03451 Related to 2-Nitropropane dioxygenase Denitrification of nitroalkanes to their corresponding carbonyl compounds ARL um11382 Related to 2-Nitropropane dioxygenase Denitrification of nitroalkanes to their corresponding carbonyl compounds SKL um11901 Rel. to Lactamase, beta 2 Unknown SAL um00045 Related to soluble Quinone reductase Detoxification of quinones HKL um11449 Related to Phenol 2-monooxygenase Detoxification of phenol compounds TRL um10538 Related to 2-Dehydropantoate-2 reductase Panthotenate (vitamin b5) synthesis ANL um02920 Histidine phosphatase domain containing p. Unknown TKL um01941 Uncharacterized α/β hydrolase Unknown TKL um02502 Uncharacterized α/β hydrolase Unknown SKL um12002 Uncharacterized α/β hydrolase Unknown AKL um06455 Uncharacterized α/β hydrolase Unknown SRL um04482 Uncharacterized α/β hydrolase Unknown, “toxin biosynthesis protein” SRL um01771 Uncharacterized protein (CoA-binding fold) Unknown GKL um12171 Uncharacterized protein Unknown SKL um03250 Uncharacterized protein Unknown SKL um12308 Uncharacterized protein Unknown ANL um11300 Uncharacterized protein Unknown SKL um02990 Hydroxysteroid dehydrogenase-like 2 Unknown Miscellaneous peroxisomal membrane proteins um03207 Rel. to MPV17-like 2 Unknown n.a. n.a. um03019 Rel. to MPV17-like 2 Unknown n.a. n.a. um05390 Rel. to PXMP4 (PMP24) Unknown n.a. n.a. um00984 Rel. to Mosc2 Reduction of N-hydroxylated drugs in mitochondria, Po function unknown n.a. n.a. um01257 Related to ANT1 - Peroxisomal ATP carrier Import/export of ATP/AMP n.a. n.a. um04444 Related to ANT1 – Peroxisomal ATP carrier Import/export of ATP/AMP n.a. n.a. um01782 Related PMP47/PMP34 Import/export of ATP/AMP n.a. n.a. um02959 Rel. to ACBD5 Pexophagy n.a. n.a. Proteins with a conserved PTS1 found only in Ustilaginomycetes** um11593 Related to PYC2 - pyruvate carboxylase 2 Gluconeogenesis: ATP + pyruvate + HCO3- = ADP + P + oxaloacetate PHL um05747 Related to SEC31 - Component of the COPII coat of ER-Golgi vesicles SRL um10552 Related to Prolyl endopeptidase Unknown SRL um00952 Related to RTS2/Kin17 Basic zinc-finger protein, DNA/RNA-binding SRF um00037 Sim. to Monodehydroascorbate reductase Detoxification of organic radical compounds AKI um00438 Related to 50S ribosomal protein L35 Translational regulation PSL um00589 Similar to Glyoxalase Removal of the toxic metabolite methylglyoxal and related compounds ANI um00740 Related to 2-Dehydropantoate 2-reductase Panthotenate (vitamin b5) synthesis GRL um11303 Related to paraoxonase Protection from lipid peroxidation CKL um10872 Rleated to Nudix hydrolase Nudt5 hydrolysis of ADP-to AMP and ribose-5-phosphate GRL um01904 Related to Succinyl-coa ligase 2 β-chain Ligation of succinate and CoA to succinyl-CoA (tricarboxylic acid cycle) GHL um03128 Uncharacterized α/β hydrolase Unknown AHL um03699 Prob. Phosphopantothenoylcysteine synth. Catalyzes the first step in the biosynthesis of coenzyme A ARI um15025 Prob. NADP-depndt. alcohol dehydrogenase Alcohol metabolism ANL um12289 Probable Dephospho-CoA kinase Catalyzes the fifth step in the biosynthesis of coenzyme A from vitamin B5 SKL um11572 Related to Rab geranylgeranyltransferase α Transfer of a geranylgeranyl moeiety to Rab proteins AHF um10945 Uncharacterized α/β hydrolase Unknown PKL um10582 Uncharacterized α/β hydrolase Unknown PKL um03638 Uncharacterized protein Unknown ARF um04796 Uncharacterized protein Unknown SHF PTS1/2, peroxisomal targeting signal 1/2; MTS, mitochondrial targeting signal; n.a. - not applicable - predicted strong targeting signal, - predicted weak targeting signal *ADHAP, the key enzyme of peroxisomal ether lipid synthesis is not found in fungi, whereas homologs of all remaining proteins of the peroxisomal part of this pathway were identified ** Proteins with an often unusual PTS1 only found among Ustilaginomycetes. The targeting signal of many proteins in this group may not be functional