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Identification of glutathione S-transferase (GST) genes from a dark septate endophytic fungus (Exophiala pisciphila) and their expression patterns under varied metals stress.

Shen M, Zhao DK, Qiao Q, Liu L, Wang JL, Cao GH, Li T, Zhao ZW - PLoS ONE (2015)

Bottom Line: Moreover, the variable expression patterns of these EpGSTs were observed under different heavy metal stresses at their effective concentrations for inhibiting growth by 50% (EC50).These results indicate that E.Pisciphila harbored a diverse of GST genes and the up-regulated EpGSTs are closely related to the heavy metal tolerance of E. pisciphila.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Conservation and Utilization for Bioresources and Key Laboratory of Microbial Diversity in Southwest China, Ministry of Education, Yunnan University, Kunming, Yunnan, China.

ABSTRACT
Glutathione S-transferases (GSTs) compose a family of multifunctional enzymes that play important roles in the detoxification of xenobiotics and the oxidative stress response. In the present study, twenty four GST genes from the transcriptome of a metal-tolerant dark septate endophyte (DSE), Exophiala pisciphila, were identified based on sequence homology, and their responses to various heavy metal exposures were also analyzed. Phylogenetic analysis showed that the 24 GST genes from E. pisciphila (EpGSTs) were divided into eight distinct classes, including seven cytosolic classes and one mitochondrial metaxin 1-like class. Moreover, the variable expression patterns of these EpGSTs were observed under different heavy metal stresses at their effective concentrations for inhibiting growth by 50% (EC50). Lead (Pb) exposure caused the up-regulation of all EpGSTs, while cadmium (Cd), copper (Cu) and zinc (Zn) treatments led to the significant up-regulation of most of the EpGSTs (p < 0.05 to p < 0.001). Furthermore, although heavy metal-specific differences in performance were observed under various heavy metals in Escherichia coli BL21 (DE3) transformed with EpGSTN-31, the over-expression of this gene was able to enhance the heavy metal tolerance of the host cells. These results indicate that E. Pisciphila harbored a diverse of GST genes and the up-regulated EpGSTs are closely related to the heavy metal tolerance of E. pisciphila. The study represents the first investigation of the GST family in E. pisciphila and provides a primary interpretation of heavy metal detoxification for E. pisciphila.

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Phylogenetic relationships among the 24 EpGSTs.Seventy fungal proteins and seven additional full-length proteins come from plants, animals, bacteria and nematodes. The accession numbers of all these genes are listed in S1 Table. The tree begins with the GST of the N-2 class from Aspergillus flavu and extends clockwise. The amino acid sequences were aligned using Clustal W, and a UPGMA tree was generated using the MEGA 5.0 software with 1000 bootstrap replicates. The numbers at each node of the phylogenetic tree represent the bootstrap values, and only those bootstrap values greater than 45% are shown. The red letters represent the GSTs from E. pisciphila. The eight clades harboring the EpGSTs are shaded in different colors.
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pone.0123418.g001: Phylogenetic relationships among the 24 EpGSTs.Seventy fungal proteins and seven additional full-length proteins come from plants, animals, bacteria and nematodes. The accession numbers of all these genes are listed in S1 Table. The tree begins with the GST of the N-2 class from Aspergillus flavu and extends clockwise. The amino acid sequences were aligned using Clustal W, and a UPGMA tree was generated using the MEGA 5.0 software with 1000 bootstrap replicates. The numbers at each node of the phylogenetic tree represent the bootstrap values, and only those bootstrap values greater than 45% are shown. The red letters represent the GSTs from E. pisciphila. The eight clades harboring the EpGSTs are shaded in different colors.

Mentions: A total of 24 non-redundant gene loci were predicted to code for putative full-length GST proteins in E. pisciphila. The ORF, protein lengths and modular domain structure of all the 24 GST genes are provided in S1 Fig and S2 Table. The proteins included in the Ure2p-like, N-2 (GST_N family, unknown subfamily 2), N-3 (GST_N family, unknown subfamily 3), zeta, theta, GTT1, EF1Bγ, and metaxin 1-like classes were designated as EpUre2p, EpGSTN-2, EpGSTN-3, EpGSTZ, EpGSTT, EpGSTG, EpEF1Bγ and EpMetaxin1, respectively (Fig 1). Twenty three full-length genes encoding putative cytosolic GSTs and one encoding a mitochondrial GST (EpMetaxin11) were identified. These newly identified members were named according to their sequence identities, and all of the EpGST sequences have been deposited into GenBank and assigned accession numbers (S2 Table).


Identification of glutathione S-transferase (GST) genes from a dark septate endophytic fungus (Exophiala pisciphila) and their expression patterns under varied metals stress.

Shen M, Zhao DK, Qiao Q, Liu L, Wang JL, Cao GH, Li T, Zhao ZW - PLoS ONE (2015)

Phylogenetic relationships among the 24 EpGSTs.Seventy fungal proteins and seven additional full-length proteins come from plants, animals, bacteria and nematodes. The accession numbers of all these genes are listed in S1 Table. The tree begins with the GST of the N-2 class from Aspergillus flavu and extends clockwise. The amino acid sequences were aligned using Clustal W, and a UPGMA tree was generated using the MEGA 5.0 software with 1000 bootstrap replicates. The numbers at each node of the phylogenetic tree represent the bootstrap values, and only those bootstrap values greater than 45% are shown. The red letters represent the GSTs from E. pisciphila. The eight clades harboring the EpGSTs are shaded in different colors.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401685&req=5

pone.0123418.g001: Phylogenetic relationships among the 24 EpGSTs.Seventy fungal proteins and seven additional full-length proteins come from plants, animals, bacteria and nematodes. The accession numbers of all these genes are listed in S1 Table. The tree begins with the GST of the N-2 class from Aspergillus flavu and extends clockwise. The amino acid sequences were aligned using Clustal W, and a UPGMA tree was generated using the MEGA 5.0 software with 1000 bootstrap replicates. The numbers at each node of the phylogenetic tree represent the bootstrap values, and only those bootstrap values greater than 45% are shown. The red letters represent the GSTs from E. pisciphila. The eight clades harboring the EpGSTs are shaded in different colors.
Mentions: A total of 24 non-redundant gene loci were predicted to code for putative full-length GST proteins in E. pisciphila. The ORF, protein lengths and modular domain structure of all the 24 GST genes are provided in S1 Fig and S2 Table. The proteins included in the Ure2p-like, N-2 (GST_N family, unknown subfamily 2), N-3 (GST_N family, unknown subfamily 3), zeta, theta, GTT1, EF1Bγ, and metaxin 1-like classes were designated as EpUre2p, EpGSTN-2, EpGSTN-3, EpGSTZ, EpGSTT, EpGSTG, EpEF1Bγ and EpMetaxin1, respectively (Fig 1). Twenty three full-length genes encoding putative cytosolic GSTs and one encoding a mitochondrial GST (EpMetaxin11) were identified. These newly identified members were named according to their sequence identities, and all of the EpGST sequences have been deposited into GenBank and assigned accession numbers (S2 Table).

Bottom Line: Moreover, the variable expression patterns of these EpGSTs were observed under different heavy metal stresses at their effective concentrations for inhibiting growth by 50% (EC50).These results indicate that E.Pisciphila harbored a diverse of GST genes and the up-regulated EpGSTs are closely related to the heavy metal tolerance of E. pisciphila.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Conservation and Utilization for Bioresources and Key Laboratory of Microbial Diversity in Southwest China, Ministry of Education, Yunnan University, Kunming, Yunnan, China.

ABSTRACT
Glutathione S-transferases (GSTs) compose a family of multifunctional enzymes that play important roles in the detoxification of xenobiotics and the oxidative stress response. In the present study, twenty four GST genes from the transcriptome of a metal-tolerant dark septate endophyte (DSE), Exophiala pisciphila, were identified based on sequence homology, and their responses to various heavy metal exposures were also analyzed. Phylogenetic analysis showed that the 24 GST genes from E. pisciphila (EpGSTs) were divided into eight distinct classes, including seven cytosolic classes and one mitochondrial metaxin 1-like class. Moreover, the variable expression patterns of these EpGSTs were observed under different heavy metal stresses at their effective concentrations for inhibiting growth by 50% (EC50). Lead (Pb) exposure caused the up-regulation of all EpGSTs, while cadmium (Cd), copper (Cu) and zinc (Zn) treatments led to the significant up-regulation of most of the EpGSTs (p < 0.05 to p < 0.001). Furthermore, although heavy metal-specific differences in performance were observed under various heavy metals in Escherichia coli BL21 (DE3) transformed with EpGSTN-31, the over-expression of this gene was able to enhance the heavy metal tolerance of the host cells. These results indicate that E. Pisciphila harbored a diverse of GST genes and the up-regulated EpGSTs are closely related to the heavy metal tolerance of E. pisciphila. The study represents the first investigation of the GST family in E. pisciphila and provides a primary interpretation of heavy metal detoxification for E. pisciphila.

Show MeSH
Related in: MedlinePlus