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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

The quantitative expression of the CYP51 gene (mean ± SE) in mycelia grown on CM (complete medium) + methanol and CW (Chinese white pine phloem methanol extract (CWPPE)) and T (terpenoid blend) of L. qinlingensis. 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-f003: The quantitative expression of the CYP51 gene (mean ± SE) in mycelia grown on CM (complete medium) + methanol and CW (Chinese white pine phloem methanol extract (CWPPE)) and T (terpenoid blend) of L. qinlingensis. CYP expression was normalized with respect to EF1. The 2−ΔΔCt and SE values were transformed at log2 for plotting.

Mentions: To determine if L. qinlingensis CYP51F1 had a possible role in the detoxification of pine defense chemicals, we analyzed the expression profiles of CYP51F1 from mycelia grown on complete medium treated with a terpenoid blend (CM + T) or with Chinese white pine phloem methanol extract (CWPPE; CM + CW) for 12 and 36 h. Statistically-significant differences were found among treatments and time (one-way ANOVA, treatments: F = 54.536, df = 2, p < 0.001; time: F = 25.399, df = 2, p < 0.001). CYP51F1 was upregulated after being exposed to the terpene blend for 12 and 36 h, and at 36 h, the transcription level was lower than at 12 h (Figure 3). At 12 h following CWPPE treatment (CM + CW, 12 h), the expression of CYP51F1 was significantly affected compared to the methanol treatments; however, one day later (CM + CW, 36 h), the expression of the CYP51F1 gene was significantly downregulated (Figure 3).


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)

The quantitative expression of the CYP51 gene (mean ± SE) in mycelia grown on CM (complete medium) + methanol and CW (Chinese white pine phloem methanol extract (CWPPE)) and T (terpenoid blend) of L. qinlingensis. 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-f003: The quantitative expression of the CYP51 gene (mean ± SE) in mycelia grown on CM (complete medium) + methanol and CW (Chinese white pine phloem methanol extract (CWPPE)) and T (terpenoid blend) of L. qinlingensis. CYP expression was normalized with respect to EF1. The 2−ΔΔCt and SE values were transformed at log2 for plotting.
Mentions: To determine if L. qinlingensis CYP51F1 had a possible role in the detoxification of pine defense chemicals, we analyzed the expression profiles of CYP51F1 from mycelia grown on complete medium treated with a terpenoid blend (CM + T) or with Chinese white pine phloem methanol extract (CWPPE; CM + CW) for 12 and 36 h. Statistically-significant differences were found among treatments and time (one-way ANOVA, treatments: F = 54.536, df = 2, p < 0.001; time: F = 25.399, df = 2, p < 0.001). CYP51F1 was upregulated after being exposed to the terpene blend for 12 and 36 h, and at 36 h, the transcription level was lower than at 12 h (Figure 3). At 12 h following CWPPE treatment (CM + CW, 12 h), the expression of CYP51F1 was significantly affected compared to the methanol treatments; however, one day later (CM + CW, 36 h), the expression of the CYP51F1 gene was significantly downregulated (Figure 3).

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