ID   HELA_ASPFU              Reviewed;         735 AA.
AC   Q4WR16;
DT   22-FEB-2012, integrated into UniProtKB/Swiss-Prot.
DT   05-JUL-2005, sequence version 1.
DT   24-JAN-2024, entry version 97.
DE   RecName: Full=Protostadienol synthase helA {ECO:0000303|PubMed:29158519};
DE            EC=5.4.99.32 {ECO:0000269|PubMed:19216560, ECO:0000269|PubMed:19415934, ECO:0000269|PubMed:19951700, ECO:0000269|PubMed:29158519};
DE   AltName: Full=Helvolic acid biosynthesis cluster protein A {ECO:0000303|PubMed:29158519};
DE   AltName: Full=Oxidosqualene cyclase {ECO:0000303|PubMed:19415934};
DE   AltName: Full=Oxidosqualene:protostadienol cyclase {ECO:0000303|PubMed:19951700};
DE            Short=OSPC {ECO:0000303|PubMed:19951700};
GN   Name=helA {ECO:0000303|PubMed:29158519};
GN   Synonyms=osc3 {ECO:0000303|PubMed:19415934}, pdsA
GN   {ECO:0000303|PubMed:19216560}; ORFNames=AFUA_4G14770;
OS   Aspergillus fumigatus (strain ATCC MYA-4609 / CBS 101355 / FGSC A1100 /
OS   Af293) (Neosartorya fumigata).
OC   Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC   Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus;
OC   Aspergillus subgen. Fumigati.
OX   NCBI_TaxID=330879;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=ATCC MYA-4609 / CBS 101355 / FGSC A1100 / Af293;
RX   PubMed=16372009; DOI=10.1038/nature04332;
RA   Nierman W.C., Pain A., Anderson M.J., Wortman J.R., Kim H.S., Arroyo J.,
RA   Berriman M., Abe K., Archer D.B., Bermejo C., Bennett J.W., Bowyer P.,
RA   Chen D., Collins M., Coulsen R., Davies R., Dyer P.S., Farman M.L.,
RA   Fedorova N., Fedorova N.D., Feldblyum T.V., Fischer R., Fosker N.,
RA   Fraser A., Garcia J.L., Garcia M.J., Goble A., Goldman G.H., Gomi K.,
RA   Griffith-Jones S., Gwilliam R., Haas B.J., Haas H., Harris D.E.,
RA   Horiuchi H., Huang J., Humphray S., Jimenez J., Keller N., Khouri H.,
RA   Kitamoto K., Kobayashi T., Konzack S., Kulkarni R., Kumagai T., Lafton A.,
RA   Latge J.-P., Li W., Lord A., Lu C., Majoros W.H., May G.S., Miller B.L.,
RA   Mohamoud Y., Molina M., Monod M., Mouyna I., Mulligan S., Murphy L.D.,
RA   O'Neil S., Paulsen I., Penalva M.A., Pertea M., Price C., Pritchard B.L.,
RA   Quail M.A., Rabbinowitsch E., Rawlins N., Rajandream M.A., Reichard U.,
RA   Renauld H., Robson G.D., Rodriguez de Cordoba S., Rodriguez-Pena J.M.,
RA   Ronning C.M., Rutter S., Salzberg S.L., Sanchez M., Sanchez-Ferrero J.C.,
RA   Saunders D., Seeger K., Squares R., Squares S., Takeuchi M., Tekaia F.,
RA   Turner G., Vazquez de Aldana C.R., Weidman J., White O., Woodward J.R.,
RA   Yu J.-H., Fraser C.M., Galagan J.E., Asai K., Machida M., Hall N.,
RA   Barrell B.G., Denning D.W.;
RT   "Genomic sequence of the pathogenic and allergenic filamentous fungus
RT   Aspergillus fumigatus.";
RL   Nature 438:1151-1156(2005).
RN   [2]
RP   INDUCTION.
RX   PubMed=17432932; DOI=10.1371/journal.ppat.0030050;
RA   Perrin R.M., Fedorova N.D., Bok J.W., Cramer R.A., Wortman J.R., Kim H.S.,
RA   Nierman W.C., Keller N.P.;
RT   "Transcriptional regulation of chemical diversity in Aspergillus fumigatus
RT   by LaeA.";
RL   PLoS Pathog. 3:E50-E50(2007).
RN   [3]
RP   FUNCTION, CATALYTIC ACTIVITY, AND PATHWAY.
RX   PubMed=19415934; DOI=10.1021/ja8095976;
RA   Mitsuguchi H., Seshime Y., Fujii I., Shibuya M., Ebizuka Y., Kushiro T.;
RT   "Biosynthesis of steroidal antibiotic fusidanes: functional analysis of
RT   oxidosqualene cyclase and subsequent tailoring enzymes from Aspergillus
RT   fumigatus.";
RL   J. Am. Chem. Soc. 131:6402-6411(2009).
RN   [4]
RP   FUNCTION, CATALYTIC ACTIVITY, AND PATHWAY.
RX   PubMed=19216560; DOI=10.1021/ol802696a;
RA   Lodeiro S., Xiong Q., Wilson W.K., Ivanova Y., Smith M.L., May G.S.,
RA   Matsuda S.P.;
RT   "Protostadienol biosynthesis and metabolism in the pathogenic fungus
RT   Aspergillus fumigatus.";
RL   Org. Lett. 11:1241-1244(2009).
RN   [5]
RP   FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP   MUTAGENESIS OF PRO-703 AND PRO-704.
RX   PubMed=19951700; DOI=10.1016/j.bbrc.2009.11.160;
RA   Kimura M., Kushiro T., Shibuya M., Ebizuka Y., Abe I.;
RT   "Protostadienol synthase from Aspergillus fumigatus: functional conversion
RT   into lanosterol synthase.";
RL   Biochem. Biophys. Res. Commun. 391:899-902(2010).
RN   [6]
RP   INDUCTION.
RX   PubMed=25311525; DOI=10.1186/1471-2164-15-894;
RA   O'Keeffe G., Hammel S., Owens R.A., Keane T.M., Fitzpatrick D.A.,
RA   Jones G.W., Doyle S.;
RT   "RNA-seq reveals the pan-transcriptomic impact of attenuating the gliotoxin
RT   self-protection mechanism in Aspergillus fumigatus.";
RL   BMC Genomics 15:894-894(2014).
RN   [7]
RP   FUNCTION, CATALYTIC ACTIVITY, AND PATHWAY.
RX   PubMed=29158519; DOI=10.1038/s41467-017-01813-9;
RA   Lv J.M., Hu D., Gao H., Kushiro T., Awakawa T., Chen G.D., Wang C.X.,
RA   Abe I., Yao X.S.;
RT   "Biosynthesis of helvolic acid and identification of an unusual C-4-
RT   demethylation process distinct from sterol biosynthesis.";
RL   Nat. Commun. 8:1644-1644(2017).
CC   -!- FUNCTION: Protostadienol synthase; part of the gene cluster that
CC       mediates the biosynthesis of helvolic acid, an antibacterial
CC       nortriterpenoid (PubMed:19415934, PubMed:19216560, PubMed:19951700,
CC       PubMed:29158519). Protostadienol synthase helA cyclizes (3S)-
CC       oxidosqualene to (17Z)-protosta-17(20),24-dien-3-beta-ol
CC       (protostadienol)(PubMed:19415934, PubMed:19216560, PubMed:19951700,
CC       PubMed:29158519). The synthesis of protostadienol is followed by
CC       several steps of monooxygenation, dehydrogenation, and acyl transfer to
CC       yield the final helvolic acid (PubMed:19216560). Following the
CC       cyclization to the tetracyclic protostadienol by helA, cytochrome P450
CC       monooxygenases helB1-mediated and helB2-mediated oxidation at C-4 and
CC       C-16, acyltransferase helD2-dependent acetylation of 16-OH, oxidation
CC       of C-21 by cytochrome P450 monooxygenase helB4, and short chain
CC       dehydrogenase helC-dependent oxidative decarboxylation yield the
CC       fusidane skeleton (PubMed:29158519). This intermediate is further
CC       modified in three additional steps mediated by the cytochrome P450
CC       monooxygenase helB3, the acyltransferase helD1, and the 3-ketosteroid
CC       1-dehydrogenase helE to give helvolic acid (PubMed:19415934,
CC       PubMed:19216560, PubMed:29158519). Compared with the late stages in the
CC       biosynthesis of helvolic acid, enzymes involved in the early stage
CC       modifications act in a relatively strict order (PubMed:29158519). The
CC       hydroxylation of C-16 by helB1 and subsequent acetylation by helD2
CC       should occur before the helB3-mediated oxidation of C-21
CC       (PubMed:29158519). C-4 demethylation in fusidane-type antibiotics
CC       proceeds in an unusual manner though it is also achieved by oxidative
CC       decarboxylation (PubMed:19415934, PubMed:29158519). The methyl group at
CC       C-4 beta position is oxidized by helB1 and subsequently removed by the
CC       short chain dehydrogenase helC (PubMed:19415934, PubMed:29158519).
CC       {ECO:0000269|PubMed:19216560, ECO:0000269|PubMed:19415934,
CC       ECO:0000269|PubMed:19951700, ECO:0000269|PubMed:29158519}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=(S)-2,3-epoxysqualene = (17Z)-protosta-17(20),24-dien-3beta-
CC         ol; Xref=Rhea:RHEA:30987, ChEBI:CHEBI:15441, ChEBI:CHEBI:62457;
CC         EC=5.4.99.32; Evidence={ECO:0000269|PubMed:19216560,
CC         ECO:0000269|PubMed:19415934, ECO:0000269|PubMed:19951700,
CC         ECO:0000269|PubMed:29158519};
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=14.4 uM for 2,3-oxidosqualene {ECO:0000269|PubMed:19951700};
CC   -!- PATHWAY: Mycotoxin biosynthesis. {ECO:0000269|PubMed:19216560,
CC       ECO:0000269|PubMed:19415934, ECO:0000269|PubMed:29158519}.
CC   -!- INDUCTION: Expression is under the control of the secondary metabolism
CC       regulator laeA (PubMed:17432932). Expression is down-regulated when
CC       gliT is deleted and up-regulated upon exposure to exogenous gliotoxin
CC       (PubMed:25311525). {ECO:0000269|PubMed:17432932,
CC       ECO:0000269|PubMed:25311525}.
CC   -!- SIMILARITY: Belongs to the terpene cyclase/mutase family.
CC       {ECO:0000305}.
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DR   EMBL; AAHF01000005; EAL89318.1; -; Genomic_DNA.
DR   RefSeq; XP_751356.1; XM_746263.1.
DR   AlphaFoldDB; Q4WR16; -.
DR   SMR; Q4WR16; -.
DR   STRING; 330879.Q4WR16; -.
DR   EnsemblFungi; EAL89318; EAL89318; AFUA_4G14770.
DR   GeneID; 3509337; -.
DR   KEGG; afm:AFUA_4G14770; -.
DR   VEuPathDB; FungiDB:Afu4g14770; -.
DR   eggNOG; KOG0497; Eukaryota.
DR   HOGENOM; CLU_009074_2_1_1; -.
DR   InParanoid; Q4WR16; -.
DR   OMA; QMEIKRY; -.
DR   OrthoDB; 2712461at2759; -.
DR   BioCyc; MetaCyc:MONOMER-16527; -.
DR   Proteomes; UP000002530; Chromosome 4.
DR   GO; GO:0005811; C:lipid droplet; IBA:GO_Central.
DR   GO; GO:0000250; F:lanosterol synthase activity; IMP:AspGD.
DR   GO; GO:1900581; P:(17Z)-protosta-17(20),24-dien-3beta-ol biosynthetic process; IMP:AspGD.
DR   GO; GO:0006696; P:ergosterol biosynthetic process; IBA:GO_Central.
DR   GO; GO:1900812; P:helvolic acid biosynthetic process; IDA:GO_Central.
DR   GO; GO:0019748; P:secondary metabolic process; IGC:AspGD.
DR   CDD; cd02892; SQCY_1; 1.
DR   Gene3D; 1.50.10.20; -; 2.
DR   Gene3D; 6.20.120.20; -; 1.
DR   InterPro; IPR032696; SQ_cyclase_C.
DR   InterPro; IPR032697; SQ_cyclase_N.
DR   InterPro; IPR018333; Squalene_cyclase.
DR   InterPro; IPR008930; Terpenoid_cyclase/PrenylTrfase.
DR   NCBIfam; TIGR01787; squalene_cyclas; 1.
DR   PANTHER; PTHR11764:SF20; LANOSTEROL SYNTHASE; 1.
DR   PANTHER; PTHR11764; TERPENE CYCLASE/MUTASE FAMILY MEMBER; 1.
DR   Pfam; PF13243; SQHop_cyclase_C; 1.
DR   Pfam; PF13249; SQHop_cyclase_N; 1.
DR   SFLD; SFLDG01016; Prenyltransferase_Like_2; 1.
DR   SUPFAM; SSF48239; Terpenoid cyclases/Protein prenyltransferases; 2.
PE   1: Evidence at protein level;
KW   Isomerase; Lipid biosynthesis; Lipid metabolism; Reference proteome;
KW   Repeat; Steroid biosynthesis; Virulence.
FT   CHAIN           1..735
FT                   /note="Protostadienol synthase helA"
FT                   /id="PRO_0000415495"
FT   REPEAT          132..173
FT                   /note="PFTB 1"
FT   REPEAT          490..531
FT                   /note="PFTB 2"
FT   REPEAT          567..607
FT                   /note="PFTB 3"
FT   REPEAT          616..663
FT                   /note="PFTB 4"
FT   ACT_SITE        463
FT                   /note="Proton donor"
FT                   /evidence="ECO:0000305"
FT   MUTAGEN         702..708
FT                   /note="APPGGMR->NKSCAIS: Changes its
FT                   oxidosqualene:protostadienol cyclase (OSPC) activity to
FT                   oxidosqualene:lanosterol cyclase (OSLC) activity."
FT                   /evidence="ECO:0000269|PubMed:19951700"
FT   MUTAGEN         703
FT                   /note="P->K: Impairs catalytic activity; when associated
FT                   with S-704."
FT                   /evidence="ECO:0000269|PubMed:19951700"
FT   MUTAGEN         704
FT                   /note="P->S: Impairs catalytic activity; when associated
FT                   with K-703."
FT                   /evidence="ECO:0000269|PubMed:19951700"
SQ   SEQUENCE   735 AA;  83091 MW;  AA0AC92A8A6E36F8 CRC64;
     MATDSSMPGT VIGKAEFSDT KAASEFGTDL SRWRLNVDNG RHMWEYLESE DEARKRPQSF
     LEKYWLGLPY ELPARPRATC ALEAVENGWE FFKRLQTADG HWGCNDDGPL FVTSGMVIAR
     YIVGIPIDSH MKQEMCRYLL NVVNEDGGWG LFIQSPSTVF GTVMNYCMLR ILGLGPEHPA
     MAKARNTLHR LGSARATPTW GKFWLCVLGV YEWEGMVPLP PEPLLVPASL PFNPGKWWVH
     TRNVYISMSY LYGHRFSMPP NKLVQALRDE LYDIPYQQIN WPAQRTNVSA ADRLTDPTWI
     QRSFTSALTM YETFKIPFLR RRALNEALFQ IETETRNTHY LCIAPVSFAS NMLALYHAHG
     RDSHWIRGMR DRFIDPMWLC REGLAASGTN GTSLWDTALT VQATIDAGLA ARPENQAILR
     KALEFIDNSQ IREDPLGVHH VYRQPTRGAW PFSTRDQSYA VSDTTAEAVK VIVLLQRIEG
     FPSRISDERL QQAIDLILGM ENAGGGFSAY EPVRGPKFLE LLNITELYEN VMTDNLYPEC
     TSSVIMCLTT FAREYPTYRP RDIQACLSRS IDYLLRSQYP NGGWFASWGV CFTYATMFAL
     QGLACMGWNE SNCAACQRAC SFLLQHQNPD GGWGESLDTV RFKQYLPHPD GSQVTNTAYA
     VIGLLAARCG NHEAIRRGVA YLVKEQQDTG EWLPGPLEGV FAPPGGMRYP NYKFHFTLMA
     LGRYVAIHGN ECLAI
//