BOT2_BOTFU
ID BOT2_BOTFU Reviewed; 399 AA.
AC Q6WP50;
DT 08-MAR-2011, integrated into UniProtKB/Swiss-Prot.
DT 05-JUL-2004, sequence version 1.
DT 03-AUG-2022, entry version 58.
DE RecName: Full=Presilphiperfolan-8-beta-ol synthase {ECO:0000303|PubMed:19476353};
DE Short=PSPS {ECO:0000303|PubMed:19476353};
DE EC=4.2.3.74 {ECO:0000269|PubMed:19035644, ECO:0000269|PubMed:19476353};
DE AltName: Full=Botrydial synthesis protein 2 {ECO:0000303|PubMed:19035644};
DE AltName: Full=Calcineurin-dependent protein 15 {ECO:0000303|PubMed:14651630};
DE AltName: Full=Sesquiterpene cyclase BOT2;
DE AltName: Full=Sesquiterpene synthase BOT2 {ECO:0000303|PubMed:19035644};
GN Name=BOT2; Synonyms=CND15;
OS Botryotinia fuckeliana (Noble rot fungus) (Botrytis cinerea).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Leotiomycetes;
OC Helotiales; Sclerotiniaceae; Botrytis.
OX NCBI_TaxID=40559;
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.
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, INDUCTION, IDENTIFICATION BY MASS SPECTROMETRY,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND PATHWAY.
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, CATALYTIC ACTIVITY, AND PATHWAY.
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: Presilphiperfolan-8-beta-ol synthase; part of the gene
CC cluster that mediates the biosynthesis of botrydial (PubMed:14651630,
CC PubMed:19035644, PubMed:19476353). Botrydial is necessary for
CC colonization of plant tissue by the T4 strain (PubMed:19035644). It is
CC a strain-dependent virulence factor since highly aggressive strains
CC like 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-beta-
CC ol (PSP) (PubMed:19035644, PubMed:19476353). The cytochrome P450
CC monooxygenase BOT4 then catalyzes the hydroxylation at C-4 to give a
CC probotryane intermediate (PubMed:27529428, PubMed:28617493).
CC Acetylation of the hydroxyl at C-4 is carried out by the
CC acetyltransferase BOT5, followed by the combined action of the P450
CC monooxygenases BOT3 and BOT1, to yield finally the glycol, via the
CC regio- and stereospecific hydroxylations at C-10 and C-15 of the
CC 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 -!- CATALYTIC ACTIVITY:
CC Reaction=(2E,6E)-farnesyl diphosphate + H2O = diphosphate +
CC presilphiperfolan-8beta-ol; Xref=Rhea:RHEA:27954, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:60968, ChEBI:CHEBI:175763;
CC EC=4.2.3.74; Evidence={ECO:0000269|PubMed:19035644,
CC ECO:0000269|PubMed:19476353};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420; Evidence={ECO:0000250};
CC Note=Binds 3 Mg(2+) ions per subunit. {ECO:0000250};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=6.04 uM for farnesyl diphosphate {ECO:0000269|PubMed:19035644};
CC -!- PATHWAY: Secondary metabolite biosynthesis.
CC {ECO:0000269|PubMed:19035644, ECO:0000269|PubMed:19476353}.
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 -!- DOMAIN: The Asp-Asp-Xaa-Xaa-Asp/Glu (DDXXD/E) motif is important for
CC the catalytic activity, presumably through binding to Mg(2+).
CC {ECO:0000305}.
CC -!- SIMILARITY: Belongs to the terpene synthase family. {ECO:0000305}.
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DR EMBL; AY277723; AAQ16575.1; -; Genomic_DNA.
DR AlphaFoldDB; Q6WP50; -.
DR SMR; Q6WP50; -.
DR KEGG; ag:AAQ16575; -.
DR SABIO-RK; Q6WP50; -.
DR GO; GO:0000287; F:magnesium ion binding; IDA:UniProtKB.
DR GO; GO:0010334; F:sesquiterpene synthase activity; IDA:UniProtKB.
DR GO; GO:0045338; P:farnesyl diphosphate metabolic process; IDA:UniProtKB.
DR GO; GO:0051762; P:sesquiterpene biosynthetic process; IDA:UniProtKB.
DR Gene3D; 1.10.600.10; -; 1.
DR InterPro; IPR008949; Isoprenoid_synthase_dom_sf.
DR InterPro; IPR034686; Terpene_cyclase-like_2.
DR SFLD; SFLDG01020; Terpene_Cyclase_Like_2; 1.
DR SUPFAM; SSF48576; SSF48576; 1.
PE 1: Evidence at protein level;
KW Lyase; Magnesium; Metal-binding; Virulence.
FT CHAIN 1..399
FT /note="Presilphiperfolan-8-beta-ol synthase"
FT /id="PRO_0000405531"
FT REGION 1..60
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT MOTIF 141..145
FT /note="DDXXD motif"
FT /evidence="ECO:0000305"
FT COMPBIAS 11..52
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT BINDING 141
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000250|UniProtKB:Q9UR08"
FT BINDING 141
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q9UR08"
FT BINDING 239
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q9UR08"
FT BINDING 285
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q9UR08"
FT BINDING 289
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q9UR08"
FT BINDING 292
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q9UR08"
FT BINDING 373..374
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q9UR08"
SQ SEQUENCE 399 AA; 45271 MW; 4D550D28FA6A13EE CRC64;
MAIPALEPQL HDADTSSNNM SSNSTDSGYD TNSTTPLEKS EKPNTQELKQ QQLDPKRPPF
VRVPDLFGSI MSTKPVVNPN YFAAKARGDR WIARVMNFNK AVAARNSKVD LCFLASMWAP
DAPEDRLVMM LDWNHWVFLF DDQFDEGHLK EDPAAAAEEV KQTIAIMGGN APRYTAESNP
IRYVFQQCWD RLKAVSSQEM QQRWIDQHKR YFDQLLVQVD QQVGGENFTR DVEAYMDLRR
GTIGVYPAIS LSEYGAGVNV PQHVYDHPSL QECMKVSADL VTLVNDVLSY RKDLELGVDH
NLMSLLMQRD NLSAQQAVDV IGDMVNECYR RWYLALAELP SYGEKIDYNV MKFVEICRAV
AQGNLYWSFQ TGRYLGPEGH EVHETGIMYL PPAANLVVA
