ID CYPX_DOTSN Reviewed; 511 AA. AC M2YIZ5; DT 28-FEB-2018, integrated into UniProtKB/Swiss-Prot. DT 01-MAY-2013, sequence version 1. DT 07-NOV-2018, entry version 18. DE RecName: Full=Cytochrome P450 monooxygenase cypX {ECO:0000303|PubMed:23207690}; DE EC=1.-.-.- {ECO:0000305|PubMed:23207690}; DE AltName: Full=Dothistromin biosynthesis protein cypX {ECO:0000303|PubMed:23207690}; GN Name=cypX {ECO:0000303|PubMed:23207690}; GN Synonyms=cypA {ECO:0000303|PubMed:17683963}; GN ORFNames=DOTSEDRAFT_139960; OS Dothistroma septosporum (strain NZE10 / CBS 128990) (Red band needle OS blight fungus) (Mycosphaerella pini). OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; OC Dothideomycetes; Dothideomycetidae; Capnodiales; Mycosphaerellaceae; OC Dothistroma. OX NCBI_TaxID=675120; RN [1] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=NZE10 / CBS 128990; RX PubMed=23209441; DOI=10.1371/journal.pgen.1003088; RA de Wit P.J.G.M., van der Burgt A., Oekmen B., Stergiopoulos I., RA Abd-Elsalam K.A., Aerts A.L., Bahkali A.H., Beenen H.G., Chettri P., RA Cox M.P., Datema E., de Vries R.P., Dhillon B., Ganley A.R., RA Griffiths S.A., Guo Y., Hamelin R.C., Henrissat B., Kabir M.S., RA Jashni M.K., Kema G., Klaubauf S., Lapidus A., Levasseur A., RA Lindquist E., Mehrabi R., Ohm R.A., Owen T.J., Salamov A., Schwelm A., RA Schijlen E., Sun H., van den Burg H.A., van Ham R.C.H.J., Zhang S., RA Goodwin S.B., Grigoriev I.V., Collemare J., Bradshaw R.E.; RT "The genomes of the fungal plant pathogens Cladosporium fulvum and RT Dothistroma septosporum reveal adaptation to different hosts and RT lifestyles but also signatures of common ancestry."; RL PLoS Genet. 8:E1003088-E1003088(2012). RN [2] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=NZE10 / CBS 128990; RX PubMed=23236275; DOI=10.1371/journal.ppat.1003037; RA Ohm R.A., Feau N., Henrissat B., Schoch C.L., Horwitz B.A., RA Barry K.W., Condon B.J., Copeland A.C., Dhillon B., Glaser F., RA Hesse C.N., Kosti I., LaButti K., Lindquist E.A., Lucas S., RA Salamov A.A., Bradshaw R.E., Ciuffetti L., Hamelin R.C., Kema G.H.J., RA Lawrence C., Scott J.A., Spatafora J.W., Turgeon B.G., RA de Wit P.J.G.M., Zhong S., Goodwin S.B., Grigoriev I.V.; RT "Diverse lifestyles and strategies of plant pathogenesis encoded in RT the genomes of eighteen Dothideomycetes fungi."; RL PLoS Pathog. 8:E1003037-E1003037(2012). RN [3] RP FUNCTION. RX PubMed=12039746; DOI=10.1128/AEM.68.6.2885-2892.2002; RA Bradshaw R.E., Bhatnagar D., Ganley R.J., Gillman C.J., Monahan B.J., RA Seconi J.M.; RT "Dothistroma pini, a forest pathogen, contains homologs of aflatoxin RT biosynthetic pathway genes."; RL Appl. Environ. Microbiol. 68:2885-2892(2002). RN [4] RP FUNCTION. RX PubMed=16649078; DOI=10.1007/s11046-006-0240-5; RA Bradshaw R.E., Jin H., Morgan B.S., Schwelm A., Teddy O.R., RA Young C.A., Zhang S.; RT "A polyketide synthase gene required for biosynthesis of the RT aflatoxin-like toxin, dothistromin."; RL Mycopathologia 161:283-294(2006). RN [5] RP FUNCTION, AND INDUCTION. RX PubMed=17683963; DOI=10.1016/j.fgb.2007.06.005; RA Zhang S., Schwelm A., Jin H., Collins L.J., Bradshaw R.E.; RT "A fragmented aflatoxin-like gene cluster in the forest pathogen RT Dothistroma septosporum."; RL Fungal Genet. Biol. 44:1342-1354(2007). RN [6] RP REVIEW ON FUNCTION, AND PATHWAY. RX PubMed=22069571; DOI=10.3390/toxins2112680; RA Schwelm A., Bradshaw R.E.; RT "Genetics of dothistromin biosynthesis of Dothistroma septosporum: an RT update."; RL Toxins 2:2680-2698(2010). RN [7] RP FUNCTION, INDUCTION, AND PATHWAY. RX PubMed=23207690; DOI=10.1016/j.fgb.2012.11.006; RA Chettri P., Ehrlich K.C., Cary J.W., Collemare J., Cox M.P., RA Griffiths S.A., Olson M.A., de Wit P.J., Bradshaw R.E.; RT "Dothistromin genes at multiple separate loci are regulated by AflR."; RL Fungal Genet. Biol. 51:12-20(2013). RN [8] RP FUNCTION. RX PubMed=23448391; DOI=10.1111/nph.12161; RA Bradshaw R.E., Slot J.C., Moore G.G., Chettri P., de Wit P.J., RA Ehrlich K.C., Ganley A.R., Olson M.A., Rokas A., Carbone I., Cox M.P.; RT "Fragmentation of an aflatoxin-like gene cluster in a forest RT pathogen."; RL New Phytol. 198:525-535(2013). CC -!- FUNCTION: Cytochrome P450 monooxygenase; part of the fragmented CC gene cluster that mediates the biosynthesis of dothistromin CC (DOTH), a polyketide toxin very similar in structure to the CC aflatoxin precursor, versicolorin B (PubMed:12039746, CC PubMed:17683963, PubMed:22069571, PubMed:23207690, CC PubMed:23448391). The first step of the pathway is the conversion CC of acetate to norsolorinic acid (NOR) and requires the fatty acid CC synthase subunits hexA and hexB, as well as the polyketide CC synthase pksA (PubMed:16649078, PubMed:23207690). PksA combines a CC hexanoyl starter unit and 7 malonyl-CoA extender units to CC synthesize the precursor NOR (By similarity). The hexanoyl starter CC unit is provided to the acyl-carrier protein (ACP) domain by the CC fungal fatty acid synthase hexA/hexB (By similarity). The second CC step is the conversion of NOR to averantin (AVN) and requires the CC norsolorinic acid ketoreductase nor1, which catalyzes the CC dehydration of norsolorinic acid to form (1'S)-averantin CC (PubMed:23207690). The cytochrome P450 monooxygenase avnA then CC catalyzes the hydroxylation of AVN to 5'hydroxyaverantin (HAVN) CC (PubMed:23207690). The next step is performed by adhA that CC transforms HAVN to averufin (AVF) (PubMed:23207690). Averufin CC might then be converted to hydroxyversicolorone by cypX and avfA CC (PubMed:23207690). Hydroxyversicolorone is further converted CC versiconal hemiacetal acetate (VHA) by moxY (PubMed:23207690). VHA CC is then the substrate for the versiconal hemiacetal acetate CC esterase est1 to yield versiconal (VAL) (PubMed:23207690). CC Versicolorin B synthase vbsA then converts VAL to versicolorin B CC (VERB) by closing the bisfuran ring (PubMed:16649078, CC PubMed:23207690). Then, the activity of the versicolorin B CC desaturase verB leads to versicolorin A (VERA) (PubMed:23207690). CC DotB, a predicted chloroperoxidase, may perform epoxidation of the CC A-ring of VERA (PubMed:23207690). Alternatively, a cytochrome CC P450, such as cypX or avnA could catalyze this step CC (PubMed:23207690). It is also possible that another, CC uncharacterized, cytochrome P450 enzyme is responsible for this CC step (PubMed:23207690). Opening of the epoxide could potentially CC be achieved by the epoxide hydrolase epoA (PubMed:23207690). CC However, epoA seems not to be required for DOTH biosynthesis, but CC other epoxide hydrolases may have the ability to complement this CC hydrolysis (PubMed:23207690). Alternatively, opening of the CC epoxide ring could be achieved non-enzymatically CC (PubMed:23207690). The next step is the deoxygenation of ring A to CC yield the 5,8-dihydroxyanthraquinone which is most likely CC catalyzed by the NADPH dehydrogenase encoded by ver1 CC (PubMed:23207690). The last stages of DOTH biosynthesis are CC proposed to involve hydroxylation of the bisfuran CC (PubMed:23207690). OrdB and norB might have oxidative roles here CC (PubMed:23207690). An alternative possibility is that cytochrome CC P450 monoogenases such as avnA and cypX might perform these steps CC in addition to previously proposed steps (PubMed:23207690). CC {ECO:0000250|UniProtKB:Q6UEF4, ECO:0000269|PubMed:12039746, CC ECO:0000269|PubMed:16649078, ECO:0000303|PubMed:22069571, CC ECO:0000305|PubMed:17683963, ECO:0000305|PubMed:23207690, CC ECO:0000305|PubMed:23448391}. CC -!- COFACTOR: CC Name=heme; Xref=ChEBI:CHEBI:30413; CC Evidence={ECO:0000250|UniProtKB:P04798}; CC -!- PATHWAY: Mycotoxin biosynthesis. {ECO:0000303|PubMed:22069571, CC ECO:0000305|PubMed:23207690}. CC -!- SUBCELLULAR LOCATION: Membrane {ECO:0000255}; Single-pass membrane CC protein {ECO:0000255}. CC -!- INDUCTION: Expression is positively regulated by the dothistromin- CC specific transcription factor aflR (PubMed:23207690). Dothistromin CC biosynthetic proteins are co-regulated, showing a high level of CC expression at ealy exponential phase with a subsequent decline in CC older cultures (PubMed:17683963). {ECO:0000269|PubMed:17683963, CC ECO:0000269|PubMed:23207690}. CC -!- SIMILARITY: Belongs to the cytochrome P450 family. {ECO:0000305}. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; KB446546; EME38861.1; -; Genomic_DNA. DR EnsemblFungi; EME38861; EME38861; DOTSEDRAFT_139960. DR OMA; WTLMANE; -. DR OrthoDB; EOG092C1LRA; -. DR Proteomes; UP000016933; Unassembled WGS sequence. DR GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW. DR GO; GO:0020037; F:heme binding; IEA:InterPro. DR GO; GO:0005506; F:iron ion binding; IEA:InterPro. DR GO; GO:0004497; F:monooxygenase activity; IEA:UniProtKB-KW. DR GO; GO:0016705; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen; IEA:InterPro. DR Gene3D; 1.10.630.10; -; 1. DR InterPro; IPR001128; Cyt_P450. DR InterPro; IPR017972; Cyt_P450_CS. DR InterPro; IPR002401; Cyt_P450_E_grp-I. DR InterPro; IPR036396; Cyt_P450_sf. DR Pfam; PF00067; p450; 1. DR PRINTS; PR00463; EP450I. DR PRINTS; PR00385; P450. DR SUPFAM; SSF48264; SSF48264; 1. DR PROSITE; PS00086; CYTOCHROME_P450; 1. PE 2: Evidence at transcript level; KW Complete proteome; Glycoprotein; Heme; Iron; Membrane; Metal-binding; KW Monooxygenase; Oxidoreductase; Reference proteome; Transmembrane; KW Transmembrane helix. FT CHAIN 1 511 Cytochrome P450 monooxygenase cypX. FT /FTId=PRO_0000443460. FT TRANSMEM 18 38 Helical. {ECO:0000255}. FT METAL 454 454 Iron (heme axial ligand). FT {ECO:0000250|UniProtKB:P04798}. FT CARBOHYD 162 162 N-linked (GlcNAc...) asparagine. FT {ECO:0000255|PROSITE-ProRule:PRU00498}. FT CARBOHYD 407 407 N-linked (GlcNAc...) asparagine. FT {ECO:0000255|PROSITE-ProRule:PRU00498}. SQ SEQUENCE 511 AA; 56700 MW; 0E273A0F34D277E0 CRC64; MAGELYKWIM DATAGAPLPF SLALVAAAFV LYNIVSIITT AYFSPLSKIP GPWYAKLTDL RLTYSVFAGN RIYYVDSLHQ KYGPMVRIGP KEVDVADPAA AREVHRMGTV FTKAPFYRLL SPGPVDNIFN FRDQKKHSQR RKLYAKGFTL VELRKNWEST INKTISMAVQ KMKEEAANGD TELMGWWTLM ANEIVCRLTF NGGHGTVEKG IKDPFVLMLE KRKGDLAHLL KMFIPPLYYV GRVLGKVNTR MNDIFYSQEK MFKAGAGVVK SARQDKEAGE FNQNLFAKAL QEGEGDAATL TDTDIITDAG ALLLAGSDPT AISLTFLIYL VLSRPELQKQ LEEEVASIDG EVTDTVCEGL PLMNAIIDES MRLYGAAPGG LPRSPPAGGA NLGGYYIPEG TVVDTQNWTL HTDGATWKEA QTFDHTRFLP ENRLEFSEKQ KMAFNPFGQG SRQCLGIHLG RLEMRLAVAH FFRELRGVKL AKSATPESMA VVDSFVAGVP RDRRCEVTMK A //