ID BOT1_BOTFU Reviewed; 510 AA. AC Q6WP49; DT 18-JUL-2018, integrated into UniProtKB/Swiss-Prot. DT 05-JUL-2004, sequence version 1. DT 18-JUL-2018, entry version 66. DE RecName: Full=Cytochrome P450 monooxygenase BOT1 {ECO:0000303|PubMed:19035644}; DE EC=1.-.-.- {ECO:0000269|PubMed:15986930}; DE AltName: Full=Botrydial biosynthesis cluster protein 1 {ECO:0000303|PubMed:19035644}; DE AltName: Full=Calcineurin-dependent protein 5 {ECO:0000303|PubMed:14651630}; GN Name=BOT1 {ECO:0000303|PubMed:19035644}; GN Synonyms=CND5 {ECO:0000303|PubMed:14651630}, GN P450-12 {ECO:0000303|PubMed:15986930}; OS Botryotinia fuckeliana (Noble rot fungus) (Botrytis cinerea). OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Leotiomycetes; OC Helotiales; Sclerotiniaceae; Botrytis. OX NCBI_TaxID=40559 {ECO:0000312|EMBL:AAQ16576.1}; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND INDUCTION. RC STRAIN=T4; RX PubMed=14651630; DOI=10.1046/j.1365-2958.2003.03798.x; RA Viaud M., Brunet-Simon A., Brygoo Y., Pradier J.-M., Levis C.; RT "Cyclophilin A and calcineurin functions investigated by gene RT inactivation, cyclosporin A inhibition and cDNA arrays approaches in RT the phytopathogenic fungus Botrytis cinerea."; RL Mol. Microbiol. 50:1451-1465(2003). RN [2] RP FUNCTION, DISRUPTION PHENOTYPE, CATALYTIC ACTIVITY, AND PATHWAY. RX PubMed=15986930; DOI=10.1094/MPMI-18-0602; RA Siewers V., Viaud M., Jimenez-Teja D., Collado I.G., Gronover C.S., RA Pradier J.M., Tudzynski B., Tudzynski P.; RT "Functional analysis of the cytochrome P450 monooxygenase gene bcbot1 RT of Botrytis cinerea indicates that botrydial is a strain-specific RT virulence factor."; RL Mol. Plant Microbe Interact. 18:602-612(2005). RN [3] RP FUNCTION, AND INDUCTION. RX PubMed=19035644; DOI=10.1021/cb800225v; RA Pinedo C., Wang C.M., Pradier J.M., Dalmais B., Choquer M., RA Le Pecheur P., Morgant G., Collado I.G., Cane D.E., Viaud M.; RT "Sesquiterpene synthase from the botrydial biosynthetic gene cluster RT of the phytopathogen Botrytis cinerea."; RL ACS Chem. Biol. 3:791-801(2008). RN [4] RP FUNCTION. RX PubMed=19476353; DOI=10.1021/ja9021649; RA Wang C.M., Hopsn R., Lin X., Cane D.E.; RT "Biosynthesis of the sesquiterpene botrydial in Botrytis cinerea. RT Mechanism and stereochemistry of the enzymatic formation of RT presilphiperfolan-8beta-ol."; RL J. Am. Chem. Soc. 131:8360-8361(2009). RN [5] RP FUNCTION. RX PubMed=27529428; DOI=10.1021/acschembio.6b00581; RA Moraga J., Dalmais B., Izquierdo-Bueno I., Aleu J., Hanson J.R., RA Hernandez-Galan R., Viaud M., Collado I.G.; RT "Genetic and molecular basis of botrydial biosynthesis: connecting RT cytochrome P450-encoding genes to biosynthetic intermediates."; RL ACS Chem. Biol. 11:2838-2846(2016). RN [6] RP FUNCTION, AND INDUCTION. RX PubMed=27721016; DOI=10.1016/j.fgb.2016.10.003; RA Porquier A., Morgant G., Moraga J., Dalmais B., Luyten I., Simon A., RA Pradier J.M., Amselem J., Collado I.G., Viaud M.; RT "The botrydial biosynthetic gene cluster of Botrytis cinerea displays RT a bipartite genomic structure and is positively regulated by the RT putative Zn(II)2Cys6 transcription factor BcBot6."; RL Fungal Genet. Biol. 96:33-46(2016). RN [7] RP FUNCTION. RX PubMed=28617493; DOI=10.1039/c7ob01088e; RA Franco Dos Santos G., Moraga J., Takahashi J.A., Viaud M., RA Hanson J.R., Hernandez Galan R., Collado I.G.; RT "The formation of sesquiterpenoid presilphiperfolane and cameroonane RT metabolites in the Bcbot4 null mutant of Botrytis cinerea."; RL Org. Biomol. Chem. 15:5357-5363(2017). CC -!- FUNCTION: Cytochrome P450 monooxygenase; part of the gene cluster CC that mediates the biosynthesis of botrydial (PubMed:14651630, CC PubMed:15986930). Botrydial is necessary for colonization of plant CC tissue by the T4 strain (PubMed:19035644). It is a strain- CC dependent virulence factor since highly aggressive strains like CC SAS56 or B05 still retain substantial virulence when botrydial CC synthesis is impaired, since they produce also botcinic acid CC (PubMed:15986930). The first step of botrydial biosynthesis is CC performed by the sesquiterpene synthase BOT2 which catalyzes the CC cyclization of farnesyl diphosphate (FPP) to presilphiperfolan-8- CC beta-ol (PSP) (PubMed:19035644, PubMed:19476353). The cytochrome CC P450 monooxygenase BOT4 then catalyzes the hydroxylation at C-4 to CC give a probotryane intermediate (PubMed:27529428, CC PubMed:28617493). Acetylation of the hydroxyl at C-4 is carried CC out by the acetyltransferase BOT5, followed by the combined action CC of the P450 monooxygenases BOT3 and BOT1, to yield finally the CC glycol, via the regio- and stereospecific hydroxylations at C-10 CC and C-15 of the probotryane intermediates, respectively CC (PubMed:15986930, PubMed:27529428). The cleavage of the C10-C15 CC bond of probotryane skeleton is an intriguing and chemically CC important reaction, which could be mediated by some of the CC monooxygenases or by a combination of them (PubMed:27529428). It CC is possible that either BOT3 or BOT1 would oxidize either the CC 10- or the 15-hydroxy group to the hydroperoxide derivative, which CC would then undergo heterolytic fragmentation to give the CC dialdehyde botrydial (PubMed:27529428). Finally, the dehydrogenase CC BOT7 might be involved in the conversion of botrydial to CC dihydrobotrydial (PubMed:27721016). {ECO:0000269|PubMed:14651630, CC ECO:0000269|PubMed:15986930, ECO:0000269|PubMed:19035644, CC ECO:0000269|PubMed:19476353, ECO:0000269|PubMed:27529428, CC ECO:0000269|PubMed:27721016, ECO:0000269|PubMed:28617493}. CC -!- COFACTOR: CC Name=heme; Xref=ChEBI:CHEBI:30413; CC Evidence={ECO:0000250|UniProtKB:P04798}; CC -!- PATHWAY: Secondary metabolite biosynthesis. CC {ECO:0000269|PubMed:15986930}. CC -!- SUBCELLULAR LOCATION: Membrane {ECO:0000255}; Single-pass membrane CC protein {ECO:0000255}. CC -!- INDUCTION: The botrydial biosynthesis cluster genes are co- CC regulated by the Ca(2+)/calcineurin signal transduction pathway, CC which is under the control of the alpha subunit BCG1 of a CC heterotrimeric G protein (PubMed:14651630, PubMed:19035644). CC Expression of the cluster is also positively regulated by the CC cluster-specific transcription factor BOT6 (PubMed:27721016). CC {ECO:0000269|PubMed:14651630, ECO:0000269|PubMed:19035644, CC ECO:0000269|PubMed:27721016}. CC -!- DISRUPTION PHENOTYPE: Abolishes the production of 10-beta,15- CC alpha-dihydroxyprobotryane, botrydial, and of some of its CC relatives (PubMed:15986930). {ECO:0000269|PubMed:15986930}. 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; AY277723; AAQ16576.1; -; Genomic_DNA. DR ProteinModelPortal; Q6WP49; -. DR eggNOG; KOG0156; Eukaryota. DR eggNOG; COG2124; LUCA. 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 1: Evidence at protein level; KW Glycoprotein; Heme; Iron; Membrane; Metal-binding; Monooxygenase; KW Oxidoreductase; Transmembrane; Transmembrane helix; Virulence. FT CHAIN 1 510 Cytochrome P450 monooxygenase BOT1. FT /FTId=PRO_0000444642. FT TRANSMEM 16 36 Helical. {ECO:0000255}. FT METAL 454 454 Iron (heme axial ligand). FT {ECO:0000250|UniProtKB:P04798}. FT CARBOHYD 476 476 N-linked (GlcNAc...) asparagine. FT {ECO:0000255|PROSITE-ProRule:PRU00498}. SQ SEQUENCE 510 AA; 58162 MW; A591F786FC5D565C CRC64; MGLLSDVINN FLPETPLAWA ALILASFTLY SVQLVVRRLY FHPLSKIPGP FLARSRYWYE FYQDIILGGI YVKNYAALHE KYGPVLRASP DRVHVSDPDF FHEVYSSGSK YMKDPAFFQA AGGIPEALPA IVDVEYHRRR RKLINDLFSA KSMEALSHLV LKVVQNALSK AHEHHEANKV LDIQRLYTGI TIDTIMQVLC DRTLNFIDAK EEEEPPFLAT LRTFSENFFL LKHFPILIWM ALNIPKSIAQ KLIPGEFEFR ASINQWIRDR ASEHELGVEK AEDGRKTVID LLLRPEDGGR PLTHQAVEDE TYSFAFAGTH TTSHTMSMGT YYLLSHPAKL QKLRDELKPI PKNDQGLYEY KTVRSLPYLN ACIKESLRMS SPVPGILPRL VPAEGMTWRG HYLPPGTSVS SSIYSVHTDP NIFPNPEQFI PERWLANENL DHYLVVFGKG SRACIGLNVA WMETYLTFSN FFTSLNMTLF ETNEQSTDWT DCGNAMIKKH VRVKVDSLAS //