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The CYP51F1 Gene of Leptographium qinlingensis: Sequence Characteristic, Phylogeny and Transcript Levels.

Dai L, Li Z, Yu J, Ma M, Zhang R, Chen H, Pham T - Int J Mol Sci (2015)

Bottom Line: We have identified an L. qinlingensis CYP51F1 gene, and the phylogenetic analysis shows the highest homology with the 14-α-demethylase sequence from Grosmannia clavigera (a fungal associate of Dendroctonus ponderosae).The homology modeling structure of CYP51F1 is similar to the structure of the lanosterol 14-α demethylase protein of Saccharomyces cerevisiae YJM789, which has an N-terminal membrane helix 1 (MH1) and transmembrane helix 1 (TMH1).The minimal inhibitory concentrations (MIC) of terpenoid and azole fungicides (itraconazole (ITC)) and the docking of terpenoid molecules, lanosterol and ITC in the protein structure suggested that CYP51F1 may be inhibited by terpenoid molecules by competitive binding with azole fungicides.

View Article: PubMed Central - PubMed

Affiliation: College of Forestry, Northwest A&F University, Yangling 712100, China. dailulu@nwafu.edu.cn.

ABSTRACT
Leptographium qinlingensis is a fungal associate of the Chinese white pine beetle (Dendroctonus armandi) and a pathogen of the Chinese white pine (Pinus armandi) that must overcome the terpenoid oleoresin defenses of host trees. L. qinlingensis responds to monoterpene flow with abundant mechanisms that include export and the use of these compounds as a carbon source. As one of the fungal cytochrome P450 proteins (CYPs), which play important roles in general metabolism, CYP51 (lanosterol 14-α demethylase) can catalyze the biosynthesis of ergosterol and is a target for antifungal drug. We have identified an L. qinlingensis CYP51F1 gene, and the phylogenetic analysis shows the highest homology with the 14-α-demethylase sequence from Grosmannia clavigera (a fungal associate of Dendroctonus ponderosae). The transcription level of CYP51F1 following treatment with terpenes and pine phloem extracts was upregulated, while using monoterpenes as the only carbon source led to the downregulation of CYP5F1 expression. The homology modeling structure of CYP51F1 is similar to the structure of the lanosterol 14-α demethylase protein of Saccharomyces cerevisiae YJM789, which has an N-terminal membrane helix 1 (MH1) and transmembrane helix 1 (TMH1). The minimal inhibitory concentrations (MIC) of terpenoid and azole fungicides (itraconazole (ITC)) and the docking of terpenoid molecules, lanosterol and ITC in the protein structure suggested that CYP51F1 may be inhibited by terpenoid molecules by competitive binding with azole fungicides.

No MeSH data available.


A maximum likelihood tree of cytochrome P450 gene from L. qinlingensis with partial sequences was performed using the amino acidic substitution model WAG model with a Gamma (−lnL = 2883.91). The CYP51F1 from L. qinlingensis is shown with an underline. The values of the bootstrap after 500 pseudoreplicates are shown at the nodes.
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ijms-16-12014-f001: A maximum likelihood tree of cytochrome P450 gene from L. qinlingensis with partial sequences was performed using the amino acidic substitution model WAG model with a Gamma (−lnL = 2883.91). The CYP51F1 from L. qinlingensis is shown with an underline. The values of the bootstrap after 500 pseudoreplicates are shown at the nodes.

Mentions: The CYP gene set of the CYP51 family, which had bootstrap values >99%, was found by Maximun Likelihood phylogenetic (ML-phylogenetic) analysis of the putative full-length amino acid sequences (Figure 1). BLAST searches indicated that CYP genes expressed in L. qinlingensis were similar to members of the gene family CYP51 reported in other species (Table 1). The full-length sequence from the CYP51 gene shared the highest level of amino acid sequence identity with variants from the fungal species G. clavigera kw1407, N. crassa OR74A, N. tetrasperma FGSC 2509, M. thermophila ATCC 42464, T. terrestris NRRL 8126 and O. piceae UAMH 11346 (Table 1). Amino acid sequence identity between partial-length sequences within each gene ranged from 86.3%–96.9% with respect to matched GenBank sequences (inter-variant). The sequence identity between the full length sequence of each gene and GenBank reference sequences was 77.5%–91.1%. The lanosterol 14-α demethylase sequence from G. clavigera kw1407 had the highest sequence identity.


The CYP51F1 Gene of Leptographium qinlingensis: Sequence Characteristic, Phylogeny and Transcript Levels.

Dai L, Li Z, Yu J, Ma M, Zhang R, Chen H, Pham T - Int J Mol Sci (2015)

A maximum likelihood tree of cytochrome P450 gene from L. qinlingensis with partial sequences was performed using the amino acidic substitution model WAG model with a Gamma (−lnL = 2883.91). The CYP51F1 from L. qinlingensis is shown with an underline. The values of the bootstrap after 500 pseudoreplicates are shown at the nodes.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4490426&req=5

ijms-16-12014-f001: A maximum likelihood tree of cytochrome P450 gene from L. qinlingensis with partial sequences was performed using the amino acidic substitution model WAG model with a Gamma (−lnL = 2883.91). The CYP51F1 from L. qinlingensis is shown with an underline. The values of the bootstrap after 500 pseudoreplicates are shown at the nodes.
Mentions: The CYP gene set of the CYP51 family, which had bootstrap values >99%, was found by Maximun Likelihood phylogenetic (ML-phylogenetic) analysis of the putative full-length amino acid sequences (Figure 1). BLAST searches indicated that CYP genes expressed in L. qinlingensis were similar to members of the gene family CYP51 reported in other species (Table 1). The full-length sequence from the CYP51 gene shared the highest level of amino acid sequence identity with variants from the fungal species G. clavigera kw1407, N. crassa OR74A, N. tetrasperma FGSC 2509, M. thermophila ATCC 42464, T. terrestris NRRL 8126 and O. piceae UAMH 11346 (Table 1). Amino acid sequence identity between partial-length sequences within each gene ranged from 86.3%–96.9% with respect to matched GenBank sequences (inter-variant). The sequence identity between the full length sequence of each gene and GenBank reference sequences was 77.5%–91.1%. The lanosterol 14-α demethylase sequence from G. clavigera kw1407 had the highest sequence identity.

Bottom Line: We have identified an L. qinlingensis CYP51F1 gene, and the phylogenetic analysis shows the highest homology with the 14-α-demethylase sequence from Grosmannia clavigera (a fungal associate of Dendroctonus ponderosae).The homology modeling structure of CYP51F1 is similar to the structure of the lanosterol 14-α demethylase protein of Saccharomyces cerevisiae YJM789, which has an N-terminal membrane helix 1 (MH1) and transmembrane helix 1 (TMH1).The minimal inhibitory concentrations (MIC) of terpenoid and azole fungicides (itraconazole (ITC)) and the docking of terpenoid molecules, lanosterol and ITC in the protein structure suggested that CYP51F1 may be inhibited by terpenoid molecules by competitive binding with azole fungicides.

View Article: PubMed Central - PubMed

Affiliation: College of Forestry, Northwest A&F University, Yangling 712100, China. dailulu@nwafu.edu.cn.

ABSTRACT
Leptographium qinlingensis is a fungal associate of the Chinese white pine beetle (Dendroctonus armandi) and a pathogen of the Chinese white pine (Pinus armandi) that must overcome the terpenoid oleoresin defenses of host trees. L. qinlingensis responds to monoterpene flow with abundant mechanisms that include export and the use of these compounds as a carbon source. As one of the fungal cytochrome P450 proteins (CYPs), which play important roles in general metabolism, CYP51 (lanosterol 14-α demethylase) can catalyze the biosynthesis of ergosterol and is a target for antifungal drug. We have identified an L. qinlingensis CYP51F1 gene, and the phylogenetic analysis shows the highest homology with the 14-α-demethylase sequence from Grosmannia clavigera (a fungal associate of Dendroctonus ponderosae). The transcription level of CYP51F1 following treatment with terpenes and pine phloem extracts was upregulated, while using monoterpenes as the only carbon source led to the downregulation of CYP5F1 expression. The homology modeling structure of CYP51F1 is similar to the structure of the lanosterol 14-α demethylase protein of Saccharomyces cerevisiae YJM789, which has an N-terminal membrane helix 1 (MH1) and transmembrane helix 1 (TMH1). The minimal inhibitory concentrations (MIC) of terpenoid and azole fungicides (itraconazole (ITC)) and the docking of terpenoid molecules, lanosterol and ITC in the protein structure suggested that CYP51F1 may be inhibited by terpenoid molecules by competitive binding with azole fungicides.

No MeSH data available.