ID   ALT2_ALTAL              Reviewed;        1103 AA.
AC   A0A3G9HRC2;
DT   17-JUN-2020, integrated into UniProtKB/Swiss-Prot.
DT   13-FEB-2019, sequence version 1.
DT   03-AUG-2022, entry version 11.
DE   RecName: Full=Bifunctional cytochrome P450/NADPH--P450 reductase ALT2 {ECO:0000250|UniProtKB:Q9Y8G7};
DE   AltName: Full=AAL-toxin biosynthesis cluster protein 2 {ECO:0000303|Ref.1};
DE   Includes:
DE     RecName: Full=Cytochrome P450 monooxygenase {ECO:0000250|UniProtKB:Q9Y8G7};
DE              EC=1.14.14.1 {ECO:0000250|UniProtKB:Q9Y8G7};
DE   Includes:
DE     RecName: Full=NADPH--cytochrome P450 reductase {ECO:0000250|UniProtKB:Q9Y8G7};
DE              EC=1.6.2.4 {ECO:0000250|UniProtKB:Q9Y8G7};
GN   Name=ALT2 {ECO:0000303|Ref.1};
OS   Alternaria alternata (Alternaria rot fungus) (Torula alternata).
OC   Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Dothideomycetes;
OC   Pleosporomycetidae; Pleosporales; Pleosporineae; Pleosporaceae; Alternaria;
OC   Alternaria sect. Alternaria; Alternaria alternata complex.
OX   NCBI_TaxID=5599;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=As-27;
RA   Akagi Y., Akamatsu H., Takao K., Tsuge T., Kodama M.;
RT   "AAL-toxin biosynthetic genes cluster in the tomato pathotype of Alternaria
RT   alternata.";
RL   Submitted (JUN-2014) to the EMBL/GenBank/DDBJ databases.
RN   [2]
RP   FUNCTION.
RX   PubMed=18435561; DOI=10.1021/np8000514;
RA   Zhu X., Vogeler C., Du L.;
RT   "Functional complementation of fumonisin biosynthesis in FUM1-disrupted
RT   fusarium verticillioides by the AAL-toxin polyketide synthase gene ALT1
RT   from Alternaria alternata f. sp. Lycopersici.";
RL   J. Nat. Prod. 71:957-960(2008).
RN   [3]
RP   FUNCTION.
RX   PubMed=19749175; DOI=10.1128/ec.00135-09;
RA   Akagi Y., Akamatsu H., Otani H., Kodama M.;
RT   "Horizontal chromosome transfer, a mechanism for the evolution and
RT   differentiation of a plant-pathogenic fungus.";
RL   Eukaryot. Cell 8:1732-1738(2009).
RN   [4]
RP   FUNCTION.
RX   PubMed=19449880; DOI=10.1021/np900193j;
RA   Li Y., Shen Y., Zhu X., Du L.;
RT   "Introduction of the AAL-toxin polyketide synthase gene ALT1 into FUM1-
RT   disrupted Fusarium verticillioides produces metabolites with the fumonisin
RT   methylation pattern.";
RL   J. Nat. Prod. 72:1328-1330(2009).
RN   [5]
RP   FUNCTION.
RX   DOI=10.4172/2157-7471.S2-001;
RA   Kheder A.A., Akagi Y., Tsuge T., Kodama M.;
RT   "Functional analysis of the ceramide synthase gene ALT7, a homolog of the
RT   disease resistance gene Asc1, in the plant pathogen Alternaria alternata.";
RL   J. Plant Pathol. Microbiol. 2:0-0(2012).
CC   -!- FUNCTION: Bifunctional cytochrome P450/NADPH--P450 reductase; part of
CC       the gene cluster that mediates the biosynthesis of the host-selective
CC       toxins (HSTs) AAL-toxins, sphinganine-analog mycotoxins responsible for
CC       Alternaria stem canker on tomato by the tomato pathotype
CC       (PubMed:18435561, PubMed:19749175, PubMed:19449880). The biosynthesis
CC       starts with the polyketide synthase ALT1-catalyzed C-16 carbon chain
CC       assembly from one starter acetyl-CoA unit with malonyl-CoA extender
CC       units (PubMed:18435561, PubMed:19449880). ALT1 also selectively
CC       transfers methyl groups at the first and the third cycle of chain
CC       elongation for AAL toxin (PubMed:19449880). The C-16 polyketide chain
CC       is released from the enzyme by a nucleophilic attack of a carbanion,
CC       which is derived from R-carbon of glycin by decarboxylation, on the
CC       carbonyl carbon of polyketide acyl chain (Probable). This step is
CC       probably catalyzed by a pyridoxal 5'-phosphate-dependent aminoacyl
CC       transferase ALT4 (Probable). The respective functions of the other
CC       enzymes encoded by the cluster have still to be elucidated (Probable).
CC       The sphingosine N-acyltransferase-like protein ALT7 seems not to act as
CC       a resistance/self-tolerance factor against the toxin in the toxin
CC       biosynthetic gene cluster, contrary to what is expected (Ref.5).
CC       {ECO:0000269|PubMed:18435561, ECO:0000269|PubMed:19449880,
CC       ECO:0000269|PubMed:19749175, ECO:0000269|Ref.5,
CC       ECO:0000305|PubMed:19449880}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=an organic molecule + O2 + reduced [NADPH--hemoprotein
CC         reductase] = an alcohol + H(+) + H2O + oxidized [NADPH--hemoprotein
CC         reductase]; Xref=Rhea:RHEA:17149, Rhea:RHEA-COMP:11964, Rhea:RHEA-
CC         COMP:11965, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC         ChEBI:CHEBI:30879, ChEBI:CHEBI:57618, ChEBI:CHEBI:58210,
CC         ChEBI:CHEBI:142491; EC=1.14.14.1;
CC         Evidence={ECO:0000250|UniProtKB:Q9Y8G7};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=NADPH + 2 oxidized [cytochrome P450] = H(+) + NADP(+) + 2
CC         reduced [cytochrome P450]; Xref=Rhea:RHEA:24040, Rhea:RHEA-
CC         COMP:14627, Rhea:RHEA-COMP:14628, ChEBI:CHEBI:15378,
CC         ChEBI:CHEBI:55376, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC         ChEBI:CHEBI:60344; EC=1.6.2.4;
CC         Evidence={ECO:0000250|UniProtKB:Q9Y8G7};
CC   -!- COFACTOR:
CC       Name=FAD; Xref=ChEBI:CHEBI:57692;
CC         Evidence={ECO:0000250|UniProtKB:Q9Y8G7};
CC       Note=Binds 1 FAD. {ECO:0000250|UniProtKB:Q9Y8G7};
CC   -!- COFACTOR:
CC       Name=FMN; Xref=ChEBI:CHEBI:58210;
CC         Evidence={ECO:0000250|UniProtKB:Q9Y8G7};
CC       Note=Binds 1 FMN. {ECO:0000250|UniProtKB:Q9Y8G7};
CC   -!- COFACTOR:
CC       Name=heme; Xref=ChEBI:CHEBI:30413;
CC         Evidence={ECO:0000250|UniProtKB:P04798};
CC   -!- PATHWAY: Mycotoxin biosynthesis. {ECO:0000250|UniProtKB:Q9Y8G7}.
CC   -!- MISCELLANEOUS: Gene clusters encoding host-selective toxins (HSTs) are
CC       localized on conditionally dispensable chromosomes (CDCs), also called
CC       supernumerary chromosomes, where they are present in multiple copies.
CC       The CDCs are not essential for saprophytic growth but controls host-
CC       selective pathogenicity. {ECO:0000269|PubMed:19749175}.
CC   -!- SIMILARITY: In the N-terminal section; belongs to the cytochrome P450
CC       family. {ECO:0000305}.
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DR   EMBL; AB969680; BBG74264.1; -; Genomic_DNA.
DR   AlphaFoldDB; A0A3G9HRC2; -.
DR   SMR; A0A3G9HRC2; -.
DR   GO; GO:0070330; F:aromatase activity; IEA:UniProtKB-EC.
DR   GO; GO:0010181; F:FMN binding; IEA:InterPro.
DR   GO; GO:0020037; F:heme binding; IEA:InterPro.
DR   GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR   GO; GO:0003958; F:NADPH-hemoprotein reductase activity; IEA:UniProtKB-EC.
DR   GO; GO:0008152; P:metabolic process; IEA:UniProtKB-KW.
DR   Gene3D; 1.10.630.10; -; 1.
DR   Gene3D; 1.20.990.10; -; 1.
DR   Gene3D; 3.40.50.360; -; 1.
DR   Gene3D; 3.40.50.80; -; 1.
DR   InterPro; IPR023206; Bifunctional_P450_P450_red.
DR   InterPro; IPR003097; CysJ-like_FAD-binding.
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   InterPro; IPR017927; FAD-bd_FR_type.
DR   InterPro; IPR008254; Flavodoxin/NO_synth.
DR   InterPro; IPR029039; Flavoprotein-like_sf.
DR   InterPro; IPR039261; FNR_nucleotide-bd.
DR   InterPro; IPR023173; NADPH_Cyt_P450_Rdtase_alpha.
DR   InterPro; IPR001433; OxRdtase_FAD/NAD-bd.
DR   InterPro; IPR017938; Riboflavin_synthase-like_b-brl.
DR   Pfam; PF00667; FAD_binding_1; 1.
DR   Pfam; PF00258; Flavodoxin_1; 1.
DR   Pfam; PF00175; NAD_binding_1; 1.
DR   Pfam; PF00067; p450; 1.
DR   PIRSF; PIRSF000209; Bifunctional_P450_P450R; 1.
DR   PRINTS; PR00463; EP450I.
DR   PRINTS; PR00385; P450.
DR   SUPFAM; SSF48264; SSF48264; 1.
DR   SUPFAM; SSF52218; SSF52218; 1.
DR   SUPFAM; SSF52343; SSF52343; 1.
DR   SUPFAM; SSF63380; SSF63380; 1.
DR   PROSITE; PS00086; CYTOCHROME_P450; 1.
DR   PROSITE; PS51384; FAD_FR; 1.
DR   PROSITE; PS50902; FLAVODOXIN_LIKE; 1.
PE   3: Inferred from homology;
KW   Electron transport; FAD; Flavoprotein; FMN; Heme; Iron; Metal-binding;
KW   Monooxygenase; Multifunctional enzyme; NADP; Oxidoreductase; Transport.
FT   CHAIN           1..1103
FT                   /note="Bifunctional cytochrome P450/NADPH--P450 reductase
FT                   ALT2"
FT                   /id="PRO_0000449850"
FT   DOMAIN          506..648
FT                   /note="Flavodoxin-like"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00088"
FT   DOMAIN          688..932
FT                   /note="FAD-binding FR-type"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00716"
FT   REGION          16..483
FT                   /note="Cytochrome P450"
FT                   /evidence="ECO:0000250|UniProtKB:Q9Y8G7"
FT   REGION          484..1103
FT                   /note="NADPH-P-450 reductase"
FT                   /evidence="ECO:0000250|UniProtKB:Q9Y8G7"
FT   BINDING         412
FT                   /ligand="heme"
FT                   /ligand_id="ChEBI:CHEBI:30413"
FT                   /ligand_part="Fe"
FT                   /ligand_part_id="ChEBI:CHEBI:18248"
FT                   /note="axial binding residue"
FT                   /evidence="ECO:0000250|UniProtKB:P04798"
FT   BINDING         512..516
FT                   /ligand="FMN"
FT                   /ligand_id="ChEBI:CHEBI:58210"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00088"
FT   BINDING         560..563
FT                   /ligand="FMN"
FT                   /ligand_id="ChEBI:CHEBI:58210"
FT                   /evidence="ECO:0000250|UniProtKB:P14779"
FT   BINDING         591..623
FT                   /ligand="FMN"
FT                   /ligand_id="ChEBI:CHEBI:58210"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00088"
FT   SITE            278
FT                   /note="Important for catalytic activity"
FT                   /evidence="ECO:0000250|UniProtKB:P14779"
SQ   SEQUENCE   1103 AA;  122186 MW;  4F9EDCD8CCE80AE5 CRC64;
     MAISSPSQAR SVGNPSPRCV KSSTYPFLGN ILDIDPDNFT KSLGDVYMIN YGSYRDVIVT
     SRKVAQELCD ESRFCKLPGG AIDRMKRVIG NGLFTAETRD PRWQSAHRVI APLFNPMRIR
     GMMDDMRDVC EQMCLRWARF GPGVPIKICD EMTKLTLDTI ALCTVDHRFN SFYRPDGIEE
     PFAEAVVNVM TDSLIQSNLP DWINNWVRFR SMNKFNRQAD ELRHAIEELI ESRRKNPVDR
     NDLLNAMLSH EDPETGQRLS DELVVDNLLT FFIAGHETTS SLLSFCMYYL LECPDVLQKA
     RAEVHATVGT STIMPEHLSK LPYLESVLRE TLRLRDPGPG FFVKPLRDDV IAGKYFVKKD
     QSIFIVFDSV HRDPDVYGDD ADEFRPERMS QEKFDQLPPC AYKPFGNGVR ACIGRPFAMQ
     QAILAVAMIL QHFDLIKDES YKLKIHVTMT VRPIGLTMKV RPREGLRATD VNLRMHQASG
     TATPKPLASG TDGSMLTVTK NGPMHLAIVH ASNSGTSEAL AGLLASNAVD RGLGVKSISV
     ANDIVEKLPR DVPVVIITAS YNGEPSRNAA DFVSWLKSTK QHELEGVRYA VFGCGHRDWA
     SSLFAVPKLI DSLLSTNGAE QIAQMGTSDT GGSTDIYSDF EDWTTKFLFP YLNSKQEMKD
     TKLNSGADTR DVDLHVSLGK PPRVAMRKGF AAATVTKNRS LSAPGVPEKC ELELCLPDGF
     TYKAGDHLQI LPRNSSNDVQ SVLRHFHLEA ETLVSIQSAH RNKRLGLPLD VPIMASELFA
     AYVELGRTAS SRNIHVLAGL VRSDKPKIKR ILLSLANGES YKTEVLDKRI SVLDLLKQFP
     DIEISLAGFL SLLMPIRPRS YSFSSGPNWK PGFATLTYTV VGAGKLVSKA KMTEHVAMSM
     RGGLASTFLS TLSARDDLYV SLDPASPSFY DDQSVSCPII MIAAGTGIAP FIGFLQERKL
     SLAHTALLQG SKKAGPYART LLFFGCRGPA LDSLYTEELA AFEADGLVEV RRAFSRDHTA
     AGSNGCKYVD QRLAASAEEL VELWRLGARV FVCGGKKMAN NVFDVLGPLF HEADKLDQKT
     AEDDVGKWRT TLGKGRYAEE IFI