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 25-MAY-2022, entry version 75. 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 inactivation, RT cyclosporin A inhibition and cDNA arrays approaches in the phytopathogenic RT 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 of RT Botrytis cinerea indicates that botrydial is a strain-specific virulence RT 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., Le Pecheur P., RA Morgant G., Collado I.G., Cane D.E., Viaud M.; RT "Sesquiterpene synthase from the botrydial biosynthetic gene cluster of the RT 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. Mechanism RT and stereochemistry of the enzymatic formation of presilphiperfolan-8beta- RT 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 a RT bipartite genomic structure and is positively regulated by the putative RT 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., Hanson J.R., RA 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 that CC 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-dependent CC virulence factor since highly aggressive strains like SAS56 or B05 CC still retain substantial virulence when botrydial synthesis is CC impaired, since they produce also botcinic acid (PubMed:15986930). The CC first step of botrydial biosynthesis is performed by the sesquiterpene CC synthase BOT2 which catalyzes the cyclization of farnesyl diphosphate CC (FPP) to presilphiperfolan-8-beta-ol (PSP) (PubMed:19035644, CC PubMed:19476353). The cytochrome P450 monooxygenase BOT4 then catalyzes CC the hydroxylation at C-4 to give a probotryane intermediate CC (PubMed:27529428, PubMed:28617493). Acetylation of the hydroxyl at C-4 CC is carried out by the acetyltransferase BOT5, followed by the combined CC action of the P450 monooxygenases BOT3 and BOT1, to yield finally the CC glycol, via the regio- and stereospecific hydroxylations at C-10 and C- CC 15 of the probotryane intermediates, respectively (PubMed:15986930, CC PubMed:27529428). The cleavage of the C10-C15 bond of probotryane CC skeleton is an intriguing and chemically important reaction, which CC could be mediated by some of the monooxygenases or by a combination of CC them (PubMed:27529428). It is possible that either BOT3 or BOT1 would CC oxidize either the 10- or the 15-hydroxy group to the hydroperoxide CC derivative, which would then undergo heterolytic fragmentation to give CC the 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-regulated by CC the Ca(2+)/calcineurin signal transduction pathway, which is under the CC control of the alpha subunit BCG1 of a heterotrimeric G protein CC (PubMed:14651630, PubMed:19035644). Expression of the cluster is also CC positively regulated by the cluster-specific transcription factor BOT6 CC (PubMed:27721016). {ECO:0000269|PubMed:14651630, CC ECO:0000269|PubMed:19035644, ECO:0000269|PubMed:27721016}. CC -!- DISRUPTION PHENOTYPE: Abolishes the production of 10-beta,15-alpha- CC dihydroxyprobotryane, botrydial, and of some of its relatives CC (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 AlphaFoldDB; Q6WP49; -. DR SMR; Q6WP49; -. DR PHI-base; PHI:438; -. 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 1: Evidence at protein level; KW Glycoprotein; Heme; Iron; Membrane; Metal-binding; Monooxygenase; KW Oxidoreductase; Transmembrane; Transmembrane helix; Virulence. FT CHAIN 1..510 FT /note="Cytochrome P450 monooxygenase BOT1" FT /id="PRO_0000444642" FT TRANSMEM 16..36 FT /note="Helical" FT /evidence="ECO:0000255" FT METAL 454 FT /note="Iron (heme axial ligand)" FT /evidence="ECO:0000250|UniProtKB:P04798" FT CARBOHYD 476 FT /note="N-linked (GlcNAc...) asparagine" FT /evidence="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 //