Supplemental Table 1. Oligonucleotides used in this study.

Oligo name / Sequence / Purpose1
Orf52-5′ / GCAACGTCGCGGCCGCGGCTGGCTCGGAATCAACGTTAAACTGCC / PCR of NCU03352
Orf52-3′ / GCAACGTCGCGGCCGCGGAAGTGAGACCATGTAGCTCCAAGTTGC / PCR of NCU03352
aod-1 forward / tcatcatggatggacagaggaa / qPCR
aod-1 reverse / GGCCCATCGACAGATTCGTA / qPCR
aod-2 forward / GGCAATGGATCTGTCCAGGTA / qPCR
aod-2 reverse / CCGGCAAATTGATTCATGCT / qPCR
aod-5 forward / GGCCCAGGAGGAAAAAAGC / qPCR
aod-5 reverse / GTTCGTCACCGCAGGTCAA / qPCR
tom40 forward / CGCCCCTCTCTTCCAAGTTT / qPCR
tom40 reverse / CAGCAAAGGCATAAGGGTTCA / qPCR
MCHA53 / gtagcatCcatatgacgggaacagaagccacggaaaagccc / PCR of aod-2 DNA-binding domain
MCHA55 / GTAGCATCGTCGACTGTAGCGTTGCGCCCAATGTCCGG / PCR of aod-2 DNA-binding domain
MCHA56 / GTAGCACTCATATGCCGGACGACGTTGGACCCGCC / PCR of aod-5 DNA-binding domain
MCHA58 / GTAGCACTCTCGAGATGGCCATTCCGGACAGATACAGCG / PCR of aod-5 DNA-binding domain
MCHA79 / CTTTGAGACCAGGCACGGACAAACTCGGTGTTTTC / Wild type EMSA probe
MCHA80 / GCTGACTGAAAACACCGAGTTTGTCCGTGCCTGGT / Wild type EMSA probe
MCHA81 / CTTTGAGACCAGGCATTTACAAACTCGGTGTTTTC / Mutant1 EMSA probe
MCHA82 / GCTGACTGAAAACACCGAGTTTGTAAATGCCTGGT / Mutant1 EMSA probe
MCHA83 / CTTTGAGACCAGGCACGGACAAACTTTTTGTTTTC / Mutant2 EMSA probe
MCHA84 / GCTGACTGAAAACAAAAAGTTTGTCCGTGCCTGGT / Mutant2 EMSA probe
MCHA85 / CTTTGAGACCAGGCATTTACAAACTTTTTGTTTTC / Mutant3 EMSA probe
MCHA86 / GCTGACTGAAAACAAAAAGTTTGTAAATGCCTGGT / Mutant3 EMSA probe
MCHA89 / CTTTGAGACCAGGCACGGACAAGCTCGGTGTTTTC / Mutant4 EMSA probe
MCHA90 / GCTGACTGAAAACACCGAGCTTGTCCGTGCCTGGT / Mutant4 EMSA probe
MCHA91 / CTTTGAGACCAGGCACGGACAACTCGGTGTTTTC / Mutant5 EMSA probe
MCHA92 / GCTGACTGAAAACACCGAGTTGTCCGTGCCTGGT / Mutant5 EMSA probe
MCHA93 / CTTTGAGACCAGGCACGGAGTTACTCGGTGTTTTC / Mutant9 EMSA probe
MCHA94 / GCTGACTGAAAACACCGAGTAACTCCGTGCCTGGT / Mutant9 EMSA probe
MCHA95 / CTTTGAGACCAGGCACGGACAAtACTCGGTGTTTTC / Mutant6 EMSA probe
MCHA96 / GCTGACTGAAAACACCGAGTATTGTCCGTGCCTGGT / Mutant6 EMSA probe
MCHA101 / CTTTGAGACCAGGCACGGACAAACTCGGTGTATTC / Mutant7 EMSA probe
MCHA102 / GCTGACTGAATACACCGAGTTTGTCCGTGCCTGGT / Mutant7 EMSA probe
MCHA103 / CTTTGAGACCAGGCACGGACAAACTCGGTGTTTC / Mutant8 EMSA probe
MCHA104 / GCTGACTGAAACACCGAGTTTGTCCGTGCCTGGT / Mutant8 EMSA probe

1 Mutants M1 through M9 are defined in Fig. 4 of the text.

Supplemental Figure Legends:

Figure 1. Alignment of AOD2 with homologues from Giberella zeae (Gz) and Saccharomyces cereviseae (Sc). Black highlight indicates residues conserved in all three proteins, grey highlights indicate residues conserved in two of the proteins. Dashed line above the aligned sequences indicates the zinc cluster region. The solid line above the alignment indicates the potential PAS domain. The open box above the alignment indicates the predicted nuclear localization signal.

Figure 2. Alignment of AOD5 with homologues from Giberella zeae (Gz) and Saccharomyces cereviseae (Sc). Black highlight indicates residues conserved in all three proteins, grey highlights indicate residues conserved in two of the proteins. Dashed line above the aligned sequences indicates the zinc cluster region. The solid line above the alignment indicates the potential PAS domain. The open box above the alignment indicates the predicted nuclear localization signal.

Figure 3. Alignment of N. crassa AOD2 and AOD5. Black highlight indicates identical residues. Grey highlight indicates similar residues.

NcAOD2 MTG-TEATEKPNGKEAGTKDITKSGSDTKPKDHHPTPADDVQKAPKKRRKVNHACLYCRR 59

GzAOD2 MTDRNPATAAKSAKQELAKTDDGKAVNRDKSSNTATSSNQKTTPAKKRRKVNHACVYCRR 60

ScRDS2 ------MSANSGVKRASKAFKTCLFCKR 22

NcAOD2 SHMTCDLERPCTRCIKRNIGHLCHDEPRDTESRKAKSVLG------TSTLHDSESQPDIG 113

GzAOD2 SHMTCDLERPCTRCIKRNIGHLCHDEPRDADSKKTKSVQSSQSIQAPSVVDESDAQSDMA 120

ScRDS2 SHVVCDKQRPCSRCVKRDIAHLCREDD------IAVPNEMPSQHESS 63

NcAOD2 RNATDKAMRPP--GFDSGMGNGSVQVAGAAAVGRGAPLQLVQPGSVAGIQASAL--GGS- 168

GzAOD2 RSSISSTMGPPPPTFDTTRQRGSKSFGG--VLGQGSPLSIVQPGQVSGLQGNELNNGGSS 178

ScRDS2 PNDNN------IQGKYANKAHTGIPSDYQNEPVNKSGST 96

NcAOD2 -MNQFAGLPDSWLTTQNHYHDMHNFHPNYMVAPEVTNEFNLLNEFL---SAGLLEESAFM 224

GzAOD2 NANQFVGFQDAWVTAQNHFHDMHSYHPNYLIAPEVTHEFNLLNDFL---HTSLLDDGGVS 235

ScRDS2 YGEELSPKLDSSLVNDTTSLLLP--QQPVFVSENVGSEFSSLNEFLSMLENPLLTQTSLS 154

NcAOD2 SDDHG--LILGANQSAVSGLPNANNNGNNASSNNKGNSSSTSGMLPPS-ATQGTSMLPPS 281

GzAOD2 SEDQQSSAFKRTSQSQSEMLP---RFGNNNNPMNAGGSNSMTNMS------GSMLPPPP 285

ScRDS2 SSSASNVHLENGSQTTQSPLEYQNDNRRDEIGVARQENRSPTIMSGSSNSISKGDKQDQE 214

NcAOD2 SDQTTAIGKPASTNLDNARDAYYLQAADPSGNDTPEERMQRLLRAKYEAGLLKPFNYILG 341

GzAOD2 NKEGKNIPRPGSVVPVDKAREYYLQAADPSGNDTPEERMGRVLRAKYDAGLLKPFNYING 345

ScRDS2 KEESRILANANENSAPTPKEQFFLTAADPSTEMTPEHRLKLVINAKLEAGLLKPYNYAKG 274

NcAOD2 YKRLSDYLDGHVSPTSKQKILKQIDRFRPKFREKIQLLTDMDLLMVEMWFERTLLEYDRV 401

GzAOD2 YTRLGTYLDSHIAASSKSKIIKTINSFRPKFREKAQALTDMELVYVEMWFEKQLMDYDRV 405

ScRDS2 YARLQDYMDKYMNQSSKQRILKPLSTIRPAFRTIARSLKDVDLVLVEESFERMLLSYDRV 334

NcAOD2 FASMAVPACCWRRTGQIFRGNKEMAELIGVPVESLRGGQIALHEILTEESNVRYWEEFGT 461

GzAOD2 FASMAVPACCWRRTGEIFRGNKEMAELIGVSVAQLRDGKIALHEILTEESMVRYWEEFGT 465

ScRDS2 FTSMSMPACLCRRTGEIYRANKEFASLVDCTVDDLRDGKLAIYELMTEESAVNFWEKYGS 394

NcAOD2 IAFDPAHDTLITACSLKNPNDDKGTKVVNCCFSFRIRRDDHKIPSLIVGNFLPHDP--- 517

GzAOD2 IAFDPAHETLLTACSLKNPSAESTHPIVKCCFSFMIRRDEHKLPALIVGNFLPHDPPTQ 524

ScRDS2 IAFDKGQKAVLTSCSLR---TKDGIRKRPCCFSFTIRRDRYNIPICIVGNFIPLS---- 446

Supplemental Fig. 1


NcAOD5 MPDDVGPAEAEVSGAVSESDNEYDETEVTTKDDDDEKMAERSVASEGVETNGDQKKKYDP 60

GzAod5 MPHEMEENSAEVSDVMSENE---DETETFIKDDD-EKMAD------QNNTEVKKKYDP 48

ScYBR239c ------MCTPDENDYKTST------DPDTSANTNHTL 25

NcAOD5 KDPLRPRRKKARRACYACQRAHLTCGDERPCQRCIKRGLAEACQDGVRKKAKYLHDAPPE 120

GzAod5 KDPLRPRRKKARRACFACQRAHLTCGDERPCQRCIKRGLADACQDGVRKKAKYLHDAPPE 108

ScYBR239c EKKKRKKRKNTNVACVNCSRLHVSCEAKRPCLRCISKGLTATCVDAPRKKSKYLAGIPNR 85

NcAOD5 ALRPVLGPNYNPAAAVSVR--NGHRHPSNAG-SDAGSSIG-TFYSQS--TQYPVFSSAAT 174

GzAod5 ALRPVLGPNYNPNTNPTPPRPNAQRQDSNASQSDSLSANGSNFFSQASTTTLPVYSTGAQ 168

ScYBR239c ELPMNIQPDLPPRKIMIPIYNNSSNSSLNVN---NMGEQQKFTSPQHIVHKAKFLSNAAD 142

NcAOD5 QLGSIPENLPFPQQSPVSPTFQPSSNPQLGSIGVSSVSSPMNSFPPALFDPSNPAIFNFN 234

GzAod5 APVGI-EGLPFNPQTSPTVFQPPIANPHP---PMNNILPAGNMDFNALFDPSNPALYNFD 224

ScYBR239c SEYSILSNIIYQDTLSNK------IP----IDILYSNTNSTSNSTIGNSS----NNS 185

NcAOD5 LEGLNFGSQYGAMEFGMLGHMSSGAAETPPRDPSMAQQGTSDVGFNP--SGVFGNGLNQF 292

GzAod5 LEGLNFGSQYAGWEFGILNKMALG-AETPPRENSMSQTPTTEANY----AALFGN-ANGN 278

ScYBR239c PTGTNTSPEETEMEK-IRQLYSEQRANIPPHPYPSSNQNVYSILLGPNSAKIVASQVNLF 244

NcAOD5 EKVYDNNTGLISDFLTLDAHSNGLYSQGNLQHGLPHAYAIPAGPTSLQSPSTENNSPQPT 352

GzAod5 ANGFD-HPMLGADFSGMDQNNQSLYAQGNLQHGLPHAYAIAAGPTSLASPSTDATASPHS 337

ScYBR239c ANHFP------LVPVDSADNSLNFKRLLPRDPSEKSSQINWDSSINQYYLNSETVTFP 296

NcAOD5 GFGFESPTATNYTGVPGAAGNQPGSQQPRAQKPKTPALGKLGPQSVLGKRQRDPSSIYEA 412

GzAod5 VAGMEGSPNHNFAGIP----TVPGAQR---HRPKT---AKAAPKSYLGKRQRDSAAIYES 387

ScYBR239c ELAIPLKRRKNHLVSVSLESCSPDAAN------IKSNVEWEHSLRYSTPMEIYTS 345

NcAOD5 VKEPFQYVASFHKLISLLQNRFSGASTISIVRSLASIRPSFMSCMKTLNRADLIFMEKSF 472

GzAod5 VKEPYPYTTGFHNMVAVLRNRLPANKLLRIAKALGEIRPSFISCTKDLTRQDLIFMEKCF 447

ScYBR239c INAPFSHTPGFHHLLVYLKHRFNQQDLVKMCRSIAEFRPIFIACSVTLTEEDMIFMEQCY 405

NcAOD5 QRALFEHEEFMHQSPSPAIACRRTGEIAAVNKEFTALTGWTKDVLLGKTLNLNANMGGTN 532

GzAod5 QRTLVEYDDFLQHCCAPTIVCRRSGEVAAVNKEFIALTGWTKEVLLGKEPNLNIN---TY 504

ScYBR239c QRTLLEYVKFIAQIGTPTCIWRRNGQISYVNEEFEILCGWTREELLN------452

NcAOD5 SDTLSISSKGGRGGIVGTTPRLKPLHPEQGTNADSQQQQSQQHKEQPQPVFLAELMDEAS 592

GzAod5 SGRSTNGSNTPDNNTQGDMPTP---RPQKATLDNS------NGRPQPVFLGELLDDDS 553

ScYBR239c ------KMTFIVEIMDDES 465

Supplemental Fig. 2 (part 1)

NcAOD5 VTQFYEDYAQLAFTHSRGT-VVRKCRLLKYRTQENMDAAAAAAAAASAPTASGGSGSSNG 651

GzAod5 VVEFYQDFAELAFEDSRGK-VQRSCRLNKYRSSQSLDIKSEHGVPK------598

ScYBR239c VRDYFKTLSKVAYRDFRGSEKMKVCRLLSP------495

NcAOD5 TVVNGGPDSSPAGKTEKERPTGVNVASNSILSNRVAKIDGEHGISKLERDGKLECSYTWT 711

GzAod5 ------DIQPG------ILSSRVTRIDSEHGISRIERDGKVECTYCWT 634

ScYBR239c ------IKG-KIIHCCCMWT 508

NcAOD5 IKRDVFDIPMIIMINFLPCYYRSHN--QLAV 740

GzAod5 IKRDVFDIPMMIIMNFLPRYLPDQGPQQLAV 665

ScYBR239c LKRDVSGLPLMILGNFMPILN------529

Supplemental Fig. 2 (part 2)

AOD2 ------MTGTEATEKPNGKEAGTKDITKSGSDTKPKDHHPTPADDVQKAPK------45

AOD5 MPDDVGPAEAEVSGAVSESDNEYDETEVTTKDDDDEKMAERSVASEGVETNGDQKKKYDP 60

AOD2 ------KRRKVNHACLYCRRSHMTCDLERPCTRCIKRNIGHLCHDEPR------DTESR 92

AOD5 KDPLRPRRKKARRACYACQRAHLTCGDERPCQRCIKRGLAEACQDGVRKKAKYLHDAPPE 120

AOD2 KAKSVLGTSTLHDSESQPDIGRNATDKAMRPPG-----FDSGMGNGSVQVAGAAAVG--- 144

AOD5 ALRPVLGPNYNPAAAVSVRNGHRHPSNAGSDAGSSIGTFYSQSTQYPVFSSAATQLGSIP 180

AOD2 ------RGAPLQLVQPGSVAGIQASALGGSMNQFAGLPDSWLTTQNHYHDMHNFHPNYM 197

AOD5 ENLPFPQQSPVSPTFQPSSNPQLGSIGVSSVSSPMNSFPPALFDPSNPAIFNFNLEGLNF 240

AOD2 VAPEVTNEFNLLNEFLS--AGLLEESAFMSDDHGLILGANQSAVSG-----LPNANNNGN 250

AOD5 GSQYGAMEFGMLGHMSSGAAETPPRDPSMAQQGTSDVGFNPSGVFGNGLNQFEKVYDNNT 300

AOD2 NASSNNKGNSSSTSGMLPPSATQG------TSMLPPSSDQTTAIGKPASTNLDN 298

AOD5 GLISDFLTLDAHSNGLYSQGNLQHGLPHAYAIPAGPTSLQSPSTENNSPQ--PTGFGFES 358

AOD2 ARDAYYLQAADPSGND--TPEERMQR------LLRAKYEAG-----LLKPFN 337

AOD5 PTATNYTGVPGAAGNQPGSQQPRAQKPKTPALGKLGPQSVLGKRQRDPSSIYEAVKEPFQ 418

AOD2 YILGYKRLSDYLDGHVSPTSKQKILKQIDRFRPKFREKIQLLTDMDLLMVEMWFERTLLE 397

AOD5 YVASFHKLISLLQNRFSGASTISIVRSLASIRPSFMSCMKTLNRADLIFMEKSFQRALFE 478

AOD2 YDRVFASMAVPACCWRRTGQIFRGNKEMAELIGVPVESLR------437

AOD5 HEEFMHQSPSPAIACRRTGEIAAVNKEFTALTGWTKDVLLGKTLNLNANMGGTNSDTLSI 538

AOD2 ---GGQ------IALHEILTEESNVRYWE 457

AOD5 SSKGGRGGIVGTTPRLKPLHPEQGTNADSQQQQSQQHKEQPQPVFLAELMDEASVTQFYE 598

AOD2 EFGTIAFDPAHDTLITACSLKNPNDDK------484

AOD5 DYAQLAFTHSRGTVVRKCRLLKYRTQENMDAAAAAAAAASAPTASGGSGSSNGTVVNGGP 658

AOD2 ------GTKVVN------CCFSFRIRRDDHK 503

AOD5 DSSPAGKTEKERPTGVNVASNSILSNRVAKIDGEHGISKLERDGKLECSYTWTIKRDVFD 718

AOD2 IPSLIVGNFLPHDP------517

AOD5 IPMIIMINFLPCYYRSHNQLAV 740

Supplemental Fig. 3

1