ID   C7016_ORYSJ             Reviewed;         505 AA.
AC   Q5Z5R4; A0A0P0WXW2; Q68YV8;
DT   29-OCT-2014, integrated into UniProtKB/Swiss-Prot.
DT   23-NOV-2004, sequence version 1.
DT   03-AUG-2022, entry version 137.
DE   RecName: Full=Ent-kaurene oxidase 2 {ECO:0000303|PubMed:15075394};
DE            Short=OsKO2 {ECO:0000303|PubMed:15075394};
DE            EC=1.14.14.86 {ECO:0000269|PubMed:22247270, ECO:0000269|PubMed:22487175};
DE   AltName: Full=Cytochrome P450 701A6 {ECO:0000303|PubMed:22247270};
DE   AltName: Full=Ent-kaurene oxidase-like 2 {ECO:0000303|PubMed:15316288};
DE            Short=OsKOL2 {ECO:0000303|PubMed:15316288};
DE   AltName: Full=OsKOS3 {ECO:0000303|PubMed:16299167};
DE   AltName: Full=Protein DWARF 35 {ECO:0000303|PubMed:15316288};
GN   Name=CYP701A6 {ECO:0000303|PubMed:22247270};
GN   Synonyms=D35 {ECO:0000303|PubMed:15316288};
GN   OrderedLocusNames=Os06g0570100 {ECO:0000312|EMBL:BAF19823.1},
GN   LOC_Os06g37364 {ECO:0000305};
GN   ORFNames=OSJNBa0062E01.38 {ECO:0000312|EMBL:BAD54598.1};
OS   Oryza sativa subsp. japonica (Rice).
OC   Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
OC   Spermatophyta; Magnoliopsida; Liliopsida; Poales; Poaceae; BOP clade;
OC   Oryzoideae; Oryzeae; Oryzinae; Oryza; Oryza sativa.
OX   NCBI_TaxID=39947;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [MRNA].
RC   STRAIN=cv. Xiushui 11;
RX   PubMed=16299167; DOI=10.1104/pp.105.072306;
RA   Zhu S., Gao F., Cao X., Chen M., Ye G., Wei C., Li Y.;
RT   "The rice dwarf virus P2 protein interacts with ent-kaurene oxidases in
RT   vivo, leading to reduced biosynthesis of gibberellins and rice dwarf
RT   symptoms.";
RL   Plant Physiol. 139:1935-1945(2005).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=cv. Nipponbare;
RX   PubMed=16100779; DOI=10.1038/nature03895;
RG   International rice genome sequencing project (IRGSP);
RT   "The map-based sequence of the rice genome.";
RL   Nature 436:793-800(2005).
RN   [3]
RP   GENOME REANNOTATION.
RC   STRAIN=cv. Nipponbare;
RX   PubMed=18089549; DOI=10.1093/nar/gkm978;
RG   The rice annotation project (RAP);
RT   "The rice annotation project database (RAP-DB): 2008 update.";
RL   Nucleic Acids Res. 36:D1028-D1033(2008).
RN   [4]
RP   GENOME REANNOTATION.
RC   STRAIN=cv. Nipponbare;
RX   PubMed=24280374; DOI=10.1186/1939-8433-6-4;
RA   Kawahara Y., de la Bastide M., Hamilton J.P., Kanamori H., McCombie W.R.,
RA   Ouyang S., Schwartz D.C., Tanaka T., Wu J., Zhou S., Childs K.L.,
RA   Davidson R.M., Lin H., Quesada-Ocampo L., Vaillancourt B., Sakai H.,
RA   Lee S.S., Kim J., Numa H., Itoh T., Buell C.R., Matsumoto T.;
RT   "Improvement of the Oryza sativa Nipponbare reference genome using next
RT   generation sequence and optical map data.";
RL   Rice 6:4-4(2013).
RN   [5]
RP   FUNCTION, TISSUE SPECIFICITY, DISRUPTION PHENOTYPE, AND MUTAGENESIS OF
RP   ARG-238.
RX   PubMed=15316288; DOI=10.1023/b:plan.0000038261.21060.47;
RA   Itoh H., Tatsumi T., Sakamoto T., Otomo K., Toyomasu T., Kitano H.,
RA   Ashikari M., Ichihara S., Matsuoka M.;
RT   "A rice semi-dwarf gene, Tan-Ginbozu (D35), encodes the gibberellin
RT   biosynthesis enzyme, ent-kaurene oxidase.";
RL   Plant Mol. Biol. 54:533-547(2004).
RN   [6]
RP   GENE FAMILY.
RX   PubMed=15075394; DOI=10.1104/pp.103.033696;
RA   Sakamoto T., Miura K., Itoh H., Tatsumi T., Ueguchi-Tanaka M., Ishiyama K.,
RA   Kobayashi M., Agrawal G.K., Takeda S., Abe K., Miyao A., Hirochika H.,
RA   Kitano H., Ashikari M., Matsuoka M.;
RT   "An overview of gibberellin metabolism enzyme genes and their related
RT   mutants in rice.";
RL   Plant Physiol. 134:1642-1653(2004).
RN   [7]
RP   FUNCTION, AND CATALYTIC ACTIVITY.
RX   PubMed=22487175; DOI=10.1016/j.bmcl.2012.03.028;
RA   Todoroki Y., Naiki K., Muramatsu T., Ohnishi T., Ueno K., Mizutani M.,
RA   Hirai N.;
RT   "A conformationally restricted uniconazole analogue as a specific inhibitor
RT   of rice ent-kaurene oxidase, CYP701A6.";
RL   Bioorg. Med. Chem. Lett. 22:3240-3243(2012).
RN   [8]
RP   FUNCTION, AND CATALYTIC ACTIVITY.
RX   PubMed=22247270; DOI=10.1104/pp.111.187518;
RA   Wang Q., Hillwig M.L., Wu Y., Peters R.J.;
RT   "CYP701A8: a rice ent-kaurene oxidase paralog diverted to more specialized
RT   diterpenoid metabolism.";
RL   Plant Physiol. 158:1418-1425(2012).
CC   -!- FUNCTION: Catalyzes three successive oxidations of the 4-methyl group
CC       of ent-kaurene giving kaurenoic acid, a key step in gibberellins (GAs)
CC       biosynthesis. GAs, which are involved many processes, including stem
CC       elongation, play a central role in plant development.
CC       {ECO:0000269|PubMed:15316288, ECO:0000269|PubMed:22247270,
CC       ECO:0000269|PubMed:22487175}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=ent-kaur-16-ene + 3 O2 + 3 reduced [NADPH--hemoprotein
CC         reductase] = ent-kaur-16-en-19-oate + 4 H(+) + 4 H2O + 3 oxidized
CC         [NADPH--hemoprotein reductase]; Xref=Rhea:RHEA:32323, Rhea:RHEA-
CC         COMP:11964, Rhea:RHEA-COMP:11965, ChEBI:CHEBI:15377,
CC         ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:15415,
CC         ChEBI:CHEBI:57297, ChEBI:CHEBI:57618, ChEBI:CHEBI:58210;
CC         EC=1.14.14.86; Evidence={ECO:0000269|PubMed:22247270,
CC         ECO:0000269|PubMed:22487175};
CC   -!- COFACTOR:
CC       Name=heme; Xref=ChEBI:CHEBI:30413;
CC         Evidence={ECO:0000250|UniProtKB:P04798};
CC   -!- PATHWAY: Plant hormone biosynthesis; gibberellin biosynthesis.
CC   -!- SUBCELLULAR LOCATION: Membrane {ECO:0000255}; Single-pass membrane
CC       protein {ECO:0000255}.
CC   -!- TISSUE SPECIFICITY: Widely expressed. {ECO:0000269|PubMed:15316288}.
CC   -!- DISRUPTION PHENOTYPE: Semi-dwarf phenotype.
CC       {ECO:0000269|PubMed:15316288}.
CC   -!- SIMILARITY: Belongs to the cytochrome P450 family.
CC       {ECO:0000255|RuleBase:RU000461}.
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DR   EMBL; AY660665; AAT81230.1; -; mRNA.
DR   EMBL; AP005471; BAD54598.1; -; Genomic_DNA.
DR   EMBL; AP008212; BAF19823.1; -; Genomic_DNA.
DR   EMBL; AP014962; BAS98310.1; -; Genomic_DNA.
DR   RefSeq; XP_015643248.1; XM_015787762.1.
DR   AlphaFoldDB; Q5Z5R4; -.
DR   SMR; Q5Z5R4; -.
DR   STRING; 4530.OS06T0570100-01; -.
DR   PRIDE; Q5Z5R4; -.
DR   EnsemblPlants; Os06t0570100-01; Os06t0570100-01; Os06g0570100.
DR   GeneID; 4341347; -.
DR   Gramene; Os06t0570100-01; Os06t0570100-01; Os06g0570100.
DR   KEGG; osa:4341347; -.
DR   eggNOG; KOG0156; Eukaryota.
DR   HOGENOM; CLU_001570_4_0_1; -.
DR   InParanoid; Q5Z5R4; -.
DR   OMA; STFMQWI; -.
DR   OrthoDB; 702827at2759; -.
DR   BRENDA; 1.14.14.86; 4460.
DR   PlantReactome; R-OSA-1119557; GA12 biosynthesis.
DR   UniPathway; UPA00390; -.
DR   Proteomes; UP000000763; Chromosome 6.
DR   Proteomes; UP000059680; Chromosome 6.
DR   Genevisible; Q5Z5R4; OS.
DR   GO; GO:0009707; C:chloroplast outer membrane; IBA:GO_Central.
DR   GO; GO:0016021; C:integral component of membrane; IEA:UniProtKB-KW.
DR   GO; GO:0052617; F:ent-kaur-16-en-19-al oxidase activity; IEA:UniProtKB-EC.
DR   GO; GO:0052616; F:ent-kaur-16-en-19-ol oxidase activity; IEA:UniProtKB-EC.
DR   GO; GO:0052615; F:ent-kaurene oxidase activity; IDA:UniProtKB.
DR   GO; GO:0020037; F:heme binding; IEA:InterPro.
DR   GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR   GO; GO:0016709; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen; IBA:GO_Central.
DR   GO; GO:0010241; P:ent-kaurene oxidation to kaurenoic acid; IDA:UniProtKB.
DR   GO; GO:0009686; P:gibberellin biosynthetic process; IMP:UniProtKB.
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   InterPro; IPR044225; KO_chloroplastic.
DR   PANTHER; PTHR47283; PTHR47283; 1.
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   Heme; Iron; Membrane; Metal-binding; Monooxygenase; NADP; Oxidoreductase;
KW   Reference proteome; Transmembrane; Transmembrane helix.
FT   CHAIN           1..505
FT                   /note="Ent-kaurene oxidase 2"
FT                   /id="PRO_0000430733"
FT   TRANSMEM        3..23
FT                   /note="Helical"
FT                   /evidence="ECO:0000255"
FT   BINDING         449
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   MUTAGEN         238
FT                   /note="R->S: In d35; semi-dwarf phenotype."
FT                   /evidence="ECO:0000269|PubMed:15316288"
FT   CONFLICT        11
FT                   /note="A -> V (in Ref. 1; AAT81230)"
FT   CONFLICT        217
FT                   /note="K -> R (in Ref. 1; AAT81230)"
SQ   SEQUENCE   505 AA;  57170 MW;  AFBE61BAFB59BF0A CRC64;
     MEAFVPGGAG AAAAAVGGFV AAAALAERAG VIAPRKRPNA PPAVPGLPII GNLHQLKEKK
     PHQTFAKWAE IYGPIYTIRT GASSVVVLNS TEVAKEAMVA KFSSISTRKL SKALTVLTRD
     KSMVATSDYC DFHKMVKRYV MSSMLGTSAQ KQFRDIRDMM IHNMLSTFHK LVKDDPHAPL
     IFRDVFKDEL FRLSMIQSLG EDVSSVYVDE FGRDISKEEI YNATVTDMMM CAIEVDWRDF
     FPYLSWVPNK SFETRVFTTE TRRTAVMRAL IKQQKERIVR GEAKTCYLDF LLAENTLTDE
     QLMMLVWEAL IEAADTTLVT TEWAMYELAK NPDKQERLYQ EIREVCGDET VTEEHLPRLP
     YLNAVFHETL RRHSPVPLIP PRFVHEDTKL AGYDVPAGTE MVINLYGCNM NRKEWESPEE
     WVPERFAGGR LEVADMYKTM AFGAGRRACA GSLQATHIAC AAVARFVQEF GWRLREGDEE
     KVDTVQLTAY KLHPLHVHLT RRGRM