P13513 · TRI5_FUSSP
- ProteinTrichodiene synthase
- GeneTRI5
- StatusUniProtKB reviewed (Swiss-Prot)
- Organism
- Amino acids374 (go to sequence)
- Protein existenceEvidence at protein level
- Annotation score5/5
Function
function
Trichodiene synthase; part of the core gene cluster that mediates the biosynthesis of trichothecenes, a very large family of chemically related bicyclic sesquiterpene compounds acting as mycotoxins, including T2-toxin (PubMed:11352533, PubMed:2817906).
The biosynthesis of trichothecenes begins with the cyclization of farnesyl diphosphate to trichodiene and is catalyzed by the trichodiene synthase TRI5 (PubMed:3800398, PubMed:7873527, PubMed:8823172).
Trichodiene undergoes a series of oxygenations catalyzed by the cytochrome P450 monooxygenase TRI4 (PubMed:7651333).
TRI4 controls the addition of four oxygens at C-2, C-3, C-11, and the C-12, C-13-epoxide to form the intermediate isotrichotriol (PubMed:16917519).
Isotrichotriol then undergoes a non-enzymatic isomerization and cyclization to form isotrichodermol (PubMed:2317042).
During this process, the oxygen at the C-2 position becomes the pyran ring oxygen and the hydroxyl group at C-11 is lost (PubMed:2317042).
More complex type A trichothecenes are built by modifying isotrichodermol through a series of paired hydroxylation and acetylation or acylation steps (PubMed:11352533).
Isotrichodermol is converted to isotrichodermin by the acetyltransferase TRI101 (PubMed:10583973).
TRI101 encodes a C-3 transacetylase that acts as a self-protection or resistance factor during biosynthesis and that the presence of a free C-3 hydroxyl group is a key component of Fusarium trichothecene phytotoxicity (PubMed:10583973).
A second hydroxyl group is added to C-15 by the trichothecene C-15 hydroxylase TRI11, producing 15-decalonectrin, which is then acetylated by TRI3, producing calonectrin (PubMed:8593041, PubMed:9435078).
A third hydroxyl group is added at C-4 by the cytochrome P450 monooxygenase TRI13, converting calonectrin to 3,15-diacetoxyspirpenol, which is subsequently acetylated by the acetyltransferase TRI7 (PubMed:11352533, PubMed:12135578).
A fourth hydroxyl group is added to C-8 by the cytochrome P450 monooxygenase TRI1, followed by the addition of an isovaleryl moiety by TRI16 (PubMed:12620849, PubMed:14532047).
Finally, the acetyl group is removed from the C-3 position by the trichothecene C-3 esterase TRI8 to produce T-2 toxin (PubMed:12039755).
The biosynthesis of trichothecenes begins with the cyclization of farnesyl diphosphate to trichodiene and is catalyzed by the trichodiene synthase TRI5 (PubMed:3800398, PubMed:7873527, PubMed:8823172).
Trichodiene undergoes a series of oxygenations catalyzed by the cytochrome P450 monooxygenase TRI4 (PubMed:7651333).
TRI4 controls the addition of four oxygens at C-2, C-3, C-11, and the C-12, C-13-epoxide to form the intermediate isotrichotriol (PubMed:16917519).
Isotrichotriol then undergoes a non-enzymatic isomerization and cyclization to form isotrichodermol (PubMed:2317042).
During this process, the oxygen at the C-2 position becomes the pyran ring oxygen and the hydroxyl group at C-11 is lost (PubMed:2317042).
More complex type A trichothecenes are built by modifying isotrichodermol through a series of paired hydroxylation and acetylation or acylation steps (PubMed:11352533).
Isotrichodermol is converted to isotrichodermin by the acetyltransferase TRI101 (PubMed:10583973).
TRI101 encodes a C-3 transacetylase that acts as a self-protection or resistance factor during biosynthesis and that the presence of a free C-3 hydroxyl group is a key component of Fusarium trichothecene phytotoxicity (PubMed:10583973).
A second hydroxyl group is added to C-15 by the trichothecene C-15 hydroxylase TRI11, producing 15-decalonectrin, which is then acetylated by TRI3, producing calonectrin (PubMed:8593041, PubMed:9435078).
A third hydroxyl group is added at C-4 by the cytochrome P450 monooxygenase TRI13, converting calonectrin to 3,15-diacetoxyspirpenol, which is subsequently acetylated by the acetyltransferase TRI7 (PubMed:11352533, PubMed:12135578).
A fourth hydroxyl group is added to C-8 by the cytochrome P450 monooxygenase TRI1, followed by the addition of an isovaleryl moiety by TRI16 (PubMed:12620849, PubMed:14532047).
Finally, the acetyl group is removed from the C-3 position by the trichothecene C-3 esterase TRI8 to produce T-2 toxin (PubMed:12039755).
Miscellaneous
Trichothecenes are sesquiterpenoid toxins that act by inhibiting protein biosynthesis.
Catalytic activity
- (2E,6E)-farnesyl diphosphate = diphosphate + trichodiene
Cofactor
Mn2+ (UniProtKB | Rhea| CHEBI:29035 )
Note: Some of the cofactor binding sites show unusual localization within the protein.
Activity regulation
Benzyl triethylammonium cation (BTAC) acts as a competitive inhibitor of trichodiene synthase reaction in the presence of pyrophosphate (PPi) (PubMed:17678871).
Kinetics
KM | SUBSTRATE | pH | TEMPERATURE[C] | NOTES | EVIDENCE | |
---|---|---|---|---|---|---|
62 nM | farnesyl pyrophosphate | |||||
78 nM | Mg2+ | |||||
84.8 nM | Mn2+ |
pH Dependence
Optimum pH is 6.75-7.75.
Pathway
Sesquiterpene biosynthesis; trichothecene biosynthesis.
Features
Showing features for binding site.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Binding site | 100 | Mg2+ 1 (UniProtKB | ChEBI) | ||||
Sequence: D | ||||||
Binding site | 164 | Mg2+ 1 (UniProtKB | ChEBI) | ||||
Sequence: E | ||||||
Binding site | 225 | Mg2+ 2 (UniProtKB | ChEBI) | ||||
Sequence: N | ||||||
Binding site | 229 | Mg2+ 2 (UniProtKB | ChEBI) | ||||
Sequence: S | ||||||
Binding site | 233 | Mg2+ 2 (UniProtKB | ChEBI) | ||||
Sequence: E | ||||||
Binding site | 239 | Mg2+ 3 (UniProtKB | ChEBI) | ||||
Sequence: D | ||||||
Binding site | 241 | Mg2+ 3 (UniProtKB | ChEBI) | ||||
Sequence: I |
GO annotations
Aspect | Term | |
---|---|---|
Molecular Function | metal ion binding | |
Molecular Function | trichodiene synthase activity | |
Biological Process | sesquiterpenoid biosynthetic process |
Keywords
- Molecular function
- Ligand
Enzyme and pathway databases
Names & Taxonomy
Protein names
- Recommended nameTrichodiene synthase
- EC number
- Alternative names
Gene names
Organism names
- Organism
- Strain
- Taxonomic lineageEukaryota > Fungi > Dikarya > Ascomycota > Pezizomycotina > Sordariomycetes > Hypocreomycetidae > Hypocreales > Nectriaceae > Fusarium
Accessions
- Primary accessionP13513
- Secondary accessions
Phenotypes & Variants
Features
Showing features for mutagenesis.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Mutagenesis | 100 | Does not significantly perturb the overall structure of trichodiene synthase but leads to an increased KM, a reduction in kcat, as well as to the production of anomalous sesquiterpene products in addition to trichodiene when incubated with farnesyl diphosphate. | ||||
Sequence: D → E | ||||||
Mutagenesis | 101 | Leads to an increased KM for Mg2+, a reduction in kcat, as well as to the production of anomalous sesquiterpene products in addition to trichodiene when incubated with farnesyl diphosphate. | ||||
Sequence: D → E | ||||||
Mutagenesis | 104 | Does not significantly affect the KM and kcat for farnesyl diphosphate. | ||||
Sequence: D → E | ||||||
Mutagenesis | 146 | Leads to the loss of activity. | ||||
Sequence: C → F | ||||||
Mutagenesis | 190 | Increases the KM for farnesyl diphosphate by about 1.3-fold and reduces the kcat by about 2000-fold. | ||||
Sequence: C → F | ||||||
Mutagenesis | 225 | Increases the KM for farnesyl diphosphate by about 6-fold and reduces the kcat by about 28-fold. Leads to complete loss of activity; when associated with S-229. | ||||
Sequence: N → D | ||||||
Mutagenesis | 229 | Increases the KM for farnesyl diphosphate by about 77-fold and reduces the kcat by about 9-fold. Leads to complete loss of activity; when associated with D-225. | ||||
Sequence: S → T | ||||||
Mutagenesis | 295 | Does not affect the catalytic activity. | ||||
Sequence: Y → F | ||||||
Mutagenesis | 304 | Does not cause large changes in the overall structure but increases the KM for farnesyl diphosphate by about 25-fold, reduces the kcat by about 200-fold, and leads to conversion of farnesyl diphosphate not only to trichodiene but to at least 2 additional C15H24 hydrocarbons. | ||||
Sequence: R → K | ||||||
Mutagenesis | 305 | Does not cause large changes in the overall structure but increases the KM for farnesyl diphosphate by about 7-fold. | ||||
Sequence: Y → F | ||||||
Mutagenesis | 305 | Increases the KM for farnesyl diphosphate by about 80-fold, reduces the kcat by about 120-fold, and leads to the conversion of farneyl diphosphate to an approximately equal mixture of trichodiene and an unidentified sesquiterpene hydrocarbon. | ||||
Sequence: Y → T |
PTM/Processing
Features
Showing features for chain.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Chain | PRO_0000221584 | 1-374 | Trichodiene synthase | |||
Sequence: MENFPTEYFLNTTVRLLEYIRYRDSNYTREERIENLHYAYNKAAHHFAQPRQQQLLKVDPKRLQASLQTIVGMVVYSWAKVSKECMADLSIHYTYTLVLDDSKDDPYPTMVNYFDDLQAGREQAHPWWALVNEHFPNVLRHFGPFCSLNLIRSTLDFFEGCWIEQYNFGGFPGSHDYPQFLRRMNGLGHCVGASLWPKEQFNERSLFLEITSAIAQMENWMVWVNDLMSFYKEFDDERDQISLVKNYVVSDEISLHEALEKLTQDTLHSSKQMVAVFSDKDPQVMDTIECFMHGYVTWHLCDRRYRLSEIYEKVKEEKTEDAQKFCKFYEQAANVGAVSPSEWAYPPVAQLANVRSKDVKEVQKPFLSSIELVE |
Expression
Induction
Expression is positively regulated by the trichothecene cluster-specific transcription activator TRI10 (PubMed:12732543).
Expression is induced between 18h and 21h growth on GYEP medium (PubMed:16347944).
The initial detection of trichothecenes occurs several hours after the initial detection of TRI5 (PubMed:16347944).
The initiation of trichothecene biosynthesis occurs with a high concentration of glucose remaining in the culture medium (PubMed:16347944).
Expression is induced between 18h and 21h growth on GYEP medium (PubMed:16347944).
The initial detection of trichothecenes occurs several hours after the initial detection of TRI5 (PubMed:16347944).
The initiation of trichothecene biosynthesis occurs with a high concentration of glucose remaining in the culture medium (PubMed:16347944).
Structure
Sequence
- Sequence statusComplete
- Length374
- Mass (Da)44,000
- Last updated1990-01-01 v1
- Checksum189E8FC1663C3763
Keywords
- Technical term
Sequence databases
Nucleotide Sequence | Protein Sequence | Molecule Type | Status | |
---|---|---|---|---|
AF364179 EMBL· GenBank· DDBJ | AAD13657.1 EMBL· GenBank· DDBJ | Genomic DNA | ||
AF359360 EMBL· GenBank· DDBJ | AAK33074.1 EMBL· GenBank· DDBJ | Genomic DNA | ||
AY032745 EMBL· GenBank· DDBJ | AAK77935.1 EMBL· GenBank· DDBJ | mRNA |