ACS_NEOBT
ID ACS_NEOBT Reviewed; 944 AA.
AC Q96WT2; Q6F5E5;
DT 05-SEP-2012, integrated into UniProtKB/Swiss-Prot.
DT 01-DEC-2001, sequence version 1.
DT 03-AUG-2022, entry version 54.
DE RecName: Full=Aphidicolan-16-beta-ol synthase {ECO:0000303|PubMed:11457369};
DE EC=4.2.3.42 {ECO:0000269|PubMed:11457369};
DE EC=5.5.1.14 {ECO:0000269|PubMed:11457369};
GN Name=ACS;
OS Neocamarosporium betae (Beet black rot fungus) (Pleospora betae).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Dothideomycetes;
OC Pleosporomycetidae; Pleosporales; Pleosporineae; Pleosporaceae;
OC Neocamarosporium.
OX NCBI_TaxID=1979465;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], CATALYTIC ACTIVITY, AND FUNCTION.
RC STRAIN=PS-13;
RX PubMed=11457369; DOI=10.1021/ja015747j;
RA Oikawa H., Toyomasu T., Toshima H., Ohashi S., Kawaide H., Kamiya Y.,
RA Ohtsuka M., Shinoda S., Mitsuhashi W., Sassa T.;
RT "Cloning and functional expression of cDNA encoding aphidicolan-16 beta-ol
RT synthase: a key enzyme responsible for formation of an unusual diterpene
RT skeleton in biosynthesis of aphidicolin.";
RL J. Am. Chem. Soc. 123:5154-5155(2001).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND IDENTIFICATION OF GENE CLUSTER.
RC STRAIN=PS-16;
RX PubMed=14745177; DOI=10.1271/bbb.68.146;
RA Toyomasu T., Nakaminami K., Toshima H., Mie T., Watanabe K., Ito H.,
RA Matsui H., Mitsuhashi W., Sassa T., Oikawa H.;
RT "Cloning of a gene cluster responsible for the biosynthesis of diterpene
RT aphidicolin, a specific inhibitor of DNA polymerase alpha.";
RL Biosci. Biotechnol. Biochem. 68:146-152(2004).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, AND PATHWAY.
RX PubMed=21897020; DOI=10.1271/bbb.110366;
RA Fujii R., Minami A., Tsukagoshi T., Sato N., Sahara T., Ohgiya S., Gomi K.,
RA Oikawa H.;
RT "Total biosynthesis of diterpene aphidicolin, a specific inhibitor of DNA
RT polymerase alpha: heterologous expression of four biosynthetic genes in
RT Aspergillus oryzae.";
RL Biosci. Biotechnol. Biochem. 75:1813-1817(2011).
CC -!- FUNCTION: Aphidicolan-16-beta-ol synthase; part of the gene cluster
CC that mediates the biosynthesis of aphidicolin, a specific inhibitor of
CC eukaryotic DNA synthesis and DNA polymerase alpha (PubMed:14745177,
CC PubMed:21897020). The geranylgeranyl pyrophosphate synthase GGS is
CC required for supplying a sufficient amount of geranylgeranyl
CC diphosphate (GGDP), the general precursor of diterpenes
CC (PubMed:21897020). The diterpene synthase ACS then catalyzes the
CC conversion of geranylgeranyl diphosphate to aphidicolan-16-beta-ol via
CC the intermediate syn-copalyldiphosphate (syn-CDP) (PubMed:11457369,
CC PubMed:21897020). In addition to aphidicolan-16-beta-ol, the enzyme
CC produces also low levels of amphidicol-15-ene and amphidicol-16-ene
CC (PubMed:11457369). The cytochrome P450 monooxygenase P450-2 then
CC catalyzes the two-step hydroxylation from aphidicolan-16-beta-ol to 3-
CC deoxyaphidicolin via a 17,3-deoxyaphidicolin intermediate
CC (PubMed:21897020). Finally, the cytochrome P450 monooxygenase P450-1
CC converts 3-deoxyaphidicolin to aphidicolin (PubMed:21897020).
CC {ECO:0000269|PubMed:11457369, ECO:0000269|PubMed:14745177,
CC ECO:0000269|PubMed:21897020}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=9alpha-copalyl diphosphate + H2O = aphidicolan-16beta-ol +
CC diphosphate; Xref=Rhea:RHEA:26213, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:29519, ChEBI:CHEBI:33019, ChEBI:CHEBI:58622; EC=4.2.3.42;
CC Evidence={ECO:0000269|PubMed:11457369};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2E,6E,10E)-geranylgeranyl diphosphate = 9alpha-copalyl
CC diphosphate; Xref=Rhea:RHEA:25524, ChEBI:CHEBI:58622,
CC ChEBI:CHEBI:58756; EC=5.5.1.14;
CC Evidence={ECO:0000269|PubMed:11457369};
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 -!- PATHWAY: Mycotoxin biosynthesis. {ECO:0000269|PubMed:21897020}.
CC -!- DOMAIN: The DXDD and DEXXE motifs are important for the catalytic
CC activity. {ECO:0000250}.
CC -!- MISCELLANEOUS: Part of a gene cluster that groups the genes involved in
CC aphidicolin biosynthesis.
CC -!- SIMILARITY: Belongs to the terpene synthase family. {ECO:0000305}.
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DR EMBL; AB049075; BAB62102.1; -; mRNA.
DR EMBL; AB114137; BAD29971.1; -; Genomic_DNA.
DR AlphaFoldDB; Q96WT2; -.
DR SMR; Q96WT2; -.
DR KEGG; ag:BAB62102; -.
DR BioCyc; MetaCyc:MON-14877; -.
DR BRENDA; 4.2.3.42; 10692.
DR GO; GO:0046567; F:aphidicolan-16 beta-ol synthase activity; IDA:UniProtKB.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0051498; F:syn-copalyl diphosphate synthase activity; IDA:UniProtKB.
DR GO; GO:0033385; P:geranylgeranyl diphosphate metabolic process; IDA:UniProtKB.
DR GO; GO:0046246; P:terpene biosynthetic process; IDA:UniProtKB.
DR Gene3D; 1.10.600.10; -; 1.
DR InterPro; IPR017057; Ent-kaurene_synthase_fun.
DR InterPro; IPR008949; Isoprenoid_synthase_dom_sf.
DR InterPro; IPR008930; Terpenoid_cyclase/PrenylTrfase.
DR PIRSF; PIRSF036498; Ent-kaurene_synthase_fungi; 1.
DR SUPFAM; SSF48239; SSF48239; 1.
PE 1: Evidence at protein level;
KW Isomerase; Lyase; Magnesium; Metal-binding.
FT CHAIN 1..944
FT /note="Aphidicolan-16-beta-ol synthase"
FT /id="PRO_0000418812"
FT MOTIF 323..326
FT /note="DXDD motif"
FT MOTIF 661..665
FT /note="DEXXE motif"
FT /evidence="ECO:0000250"
FT BINDING 323
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="4"
FT /evidence="ECO:0000250|UniProtKB:C7BKP9"
FT BINDING 325
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="4"
FT /evidence="ECO:0000250|UniProtKB:C7BKP9"
FT BINDING 661
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000250"
FT BINDING 661
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000250"
FT BINDING 665
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000250"
FT BINDING 665
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000250"
FT BINDING 835
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000250"
FT BINDING 839
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000250"
FT BINDING 843
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="3"
FT /evidence="ECO:0000250"
FT CONFLICT 445
FT /note="R -> K (in Ref. 2; BAD29971)"
FT /evidence="ECO:0000305"
FT CONFLICT 559
FT /note="S -> F (in Ref. 2; BAD29971)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 944 AA; 106581 MW; 41C781606F236CE5 CRC64;
MVPISTRSEV QDGLIAQARS LITRIVHNSD DVYGFGTLSC TVYDTAWVAL VTKHVNGIKH
WLFPESFHYI LASQCDDGTW CEDKTAQFDG VLNTIAGLLV LKRYRRDPLQ LKVDNRDLDS
RIKLATSALH SLLEEWDVST TNNVGFEIIV PTMLDLLVQE DPSLSFELKG REALTEIREA
KMNRFQPELL YQGKLMTLTH SLEALLGRID YDNVARYTVK GSMFASPSST AAYLMSASTW
DDEAETYLRY TITASTGKGS GGVPGVFPTT YFEYTWILST LFRAGFQSSE LDSPELTAMT
DTLLKAFKAF SGAIGLDSGI EPDVDDSAKI VTTLNMLGKP AHARYLCDNF EVETHFRVYP
RERDPSFSAN CNALAAFLHQ PDVEVYSSQI LKAASYLCER MWNADEKIDD KWNKSHLYSS
FLYTQVMTDL MAITEAGRLD GVFTRELLTR VCVTIFQSCL RAMLKQSHDG SWNQSLEETA
YAILHLTEAR RLCFFEQISE PLENSIYRGI TFLTTIDKPP MEYLWSDKVS YGSAYLAETY
VLAARRAAES TSVTNLVGSS IWKDNASTKM HKLVGLFHRT PLLKALPKWE LQASMIEASI
YQGLLQDARL EVLQRPKVDG GEYLSIIPFT WTSCSNSART NASASHLWEL MALSFFTYQV
DEFMEAVAGP AFKGRMTHLH AIIDEAVHCS QNRERTPGEC ENTNHVTSEL LQAVRFILDN
PTVRKASPYD RNTLLQELRI FLHAHVTQVE DNASFGRERL TGDKALTSYR SQLYRWVHTI
SSDHIAGPFC FYYATCLLGA TLTAHPPNDC FPKSSQKYLA AATCRHLSCM CRMYNDIGSW
NRDHREGNLN CLHFPEFSET TSDDAERKAS LLTLAQYERK QWCNALQQLE KEMVHGASEP
AAARLAKRRA CLIDMFCKVT DLYGQIYVLR DVSSVIKDVV RNGE
