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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.


Related in: MedlinePlus

Quantitative expression of the CYP51 gene (mean ± SE) in L. qinlingensis grown on YNB (yeast nitrogen base without amino acids) + Ma (mannose) and MT (monoterpenes) and OA (oleic acid) with different carbon sources. CYP expression was normalized with respect to EF1. The 2−ΔΔCt and SE values were transformed at log2 for plotting.
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ijms-16-12014-f004: Quantitative expression of the CYP51 gene (mean ± SE) in L. qinlingensis grown on YNB (yeast nitrogen base without amino acids) + Ma (mannose) and MT (monoterpenes) and OA (oleic acid) with different carbon sources. CYP expression was normalized with respect to EF1. The 2−ΔΔCt and SE values were transformed at log2 for plotting.

Mentions: To determine whether CYP51F1 was involved in the utilization of different carbon sources, we analyzed CYP gene expression profiles of L. qinlingensis grown on minimal medium with a single carbon source: a monoterpene blend for 10 days (yeast nitrogen base (YNB) + MT) and long-chain fatty acids (oleic acid; YNB + OA) for five days. A statistically-significant difference was found only between YNB + MT (10 days) and YNB + Ma (mannose) (3 days) (one-way ANOVA, F = 41.181, df = 1, p = 0.003). In mycelia grown on monoterpenes as the sole carbon source (YNB + MT), CYP51F1 was significantly downregulated (Figure 4). The expression of CYP51F1 displayed almost no change between YNB + OA (five days) and YNB + Ma (five days) (one-way ANOVA, F = 0.249, df = 1, p = 0.644) (Figure 4).


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)

Quantitative expression of the CYP51 gene (mean ± SE) in L. qinlingensis grown on YNB (yeast nitrogen base without amino acids) + Ma (mannose) and MT (monoterpenes) and OA (oleic acid) with different carbon sources. CYP expression was normalized with respect to EF1. The 2−ΔΔCt and SE values were transformed at log2 for plotting.
© Copyright Policy
Related In: Results  -  Collection

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

ijms-16-12014-f004: Quantitative expression of the CYP51 gene (mean ± SE) in L. qinlingensis grown on YNB (yeast nitrogen base without amino acids) + Ma (mannose) and MT (monoterpenes) and OA (oleic acid) with different carbon sources. CYP expression was normalized with respect to EF1. The 2−ΔΔCt and SE values were transformed at log2 for plotting.
Mentions: To determine whether CYP51F1 was involved in the utilization of different carbon sources, we analyzed CYP gene expression profiles of L. qinlingensis grown on minimal medium with a single carbon source: a monoterpene blend for 10 days (yeast nitrogen base (YNB) + MT) and long-chain fatty acids (oleic acid; YNB + OA) for five days. A statistically-significant difference was found only between YNB + MT (10 days) and YNB + Ma (mannose) (3 days) (one-way ANOVA, F = 41.181, df = 1, p = 0.003). In mycelia grown on monoterpenes as the sole carbon source (YNB + MT), CYP51F1 was significantly downregulated (Figure 4). The expression of CYP51F1 displayed almost no change between YNB + OA (five days) and YNB + Ma (five days) (one-way ANOVA, F = 0.249, df = 1, p = 0.644) (Figure 4).

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.


Related in: MedlinePlus