De Novo Transcriptome Sequencing of the Deep-Sea-Derived Fungus Dichotomomyces cejpii and Analysis of Gliotoxin Biosynthesis Genes

Ye, Wei; Zhang, Weiyang; Liu, Taomei; Huang, Zilei; Zhu, Muzi; Chen, Yuchan; Li, Haohua; Li, Saini

doi:10.3390/ijms19071910

Cited by 6 publications

(4 citation statements)

References 33 publications

(47 reference statements)

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“…However, the binding capacity of DcGliZ core protein with the pG promoter was indeed verified by EMSA assay. The pG promoter was obtained using genome walking kit to amplify the upstream sequences of the gliG gene, which encodes glutathione S-transferase (GST), and the function of GST in gliotoxin biosynthesis has been proven in our previous study [27]. The transcriptional activity of pG promoter was also confirmed by assaying fluorescence intensity [48].…”

Section: Discussionmentioning

confidence: 99%

Interaction of a Novel Zn2Cys6 Transcription Factor DcGliZ with Promoters in the Gliotoxin Biosynthetic Gene Cluster of the Deep-Sea-Derived Fungus Dichotomomyces cejpii

et al. 2019

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Gliotoxin is an important epipolythiodioxopiperazine, which was biosynthesized by the gli gene cluster in Aspergillus genus. However, the regulatory mechanism of gliotoxin biosynthesis remains unclear. In this study, a novel Zn2Cys6 transcription factor DcGliZ that is responsible for the regulation of gliotoxin biosynthesis from the deep-sea-derived fungus Dichotomomyces cejpii was identified. DcGliZ was expressed in Escherichia coli and effectively purified from inclusion bodies by refolding. Using electrophoretic mobility shift assay, we demonstrated that purified DcGliZ can bind to gliG, gliM, and gliN promoter regions in the gli cluster. Furthermore, the binding kinetics and affinity of DcGliZ protein with different promoters were measured by surface plasmon resonance assays, and the results demonstrated the significant interaction of DcGliZ with the gliG, gliM, and gliN promoters. These new findings would lay the foundation for the elucidation of future gliotoxin biosynthetic regulation mechanisms in D. cejpii.The putative gene cluster (gli) of gliotoxin is composed of 13 genes that encode crucial enzymes responsible for the biosynthesis of various gliotoxins and their derivatives in A. fumigatus [9]. The non-ribosomal peptide synthetase encoded by gliP gene catalyzes diketopiperazine scaffold formation, the first biosynthetic reaction in a gliotoxin biosynthesis pathway [10]. Subsequently, cytochrome P450 monooxygenase encoded by gliC catalyzes the hydroxylation at the α-carbon of L-Phe [11]. Then, glutathione S-transferase (GST) encoded by the gliG promotes the sulfurization of gliotoxin biosynthetic imtermediates; GST is known for its ability in the catalyzation of carbon-sulfur bond formation, as opposed to detoxification [12,13]. After the process by the enzymes of GliK and GliJ, the γ-glutamyl moieties are removed [14,15]. The carbon-sulfur lyase expressed by the gliI gene then catalyzes the intermediate to generate a notorious epidithiol moiety [16]. After the catalysis of cytochrome P450 monooxygenases (GliF or GliC) and GliH (function remains elusive), the N-methyltransferase encoded by gliN or gliM functions as a freestanding amide to promote amide methylation and confer stability on ETP [17,18]. Finally, the oxidoreductase GliT-mediated disulfide bridge closure might be a prerequisite for the formation of gliotoxins, and the major facilitator superfamily transporter GliA also plays an important role in exporting the toxins to prevent the harmful effect of gliotoxin on hosts [9,19].Many regulators involved in gliotoxin biosynthesis pathway have been identified in A. fumigatus, regardless of whether they deploy a non-gli cluster or a gli cluster. GtmA (or termed TtmA) is encoded outside the gli cluster and functions as a bis-thiomethyl transferase for the conversion of dithiogliotoxin to bisdethiobis (methylthio) gliotoxin (BmGT), which mainly attenuates the formation of disulfide bridge closure [20]. Other non-gli clusters encoding transcription factors, including the global regulator laeA, C 2 H 2...

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Section: Discussionmentioning

confidence: 99%

Interaction of a Novel Zn2Cys6 Transcription Factor DcGliZ with Promoters in the Gliotoxin Biosynthetic Gene Cluster of the Deep-Sea-Derived Fungus Dichotomomyces cejpii

et al. 2019

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“…The cDNA library was sequenced on the Illumina sequencing platform (Illumina HiSeq™ 2500, San Diego, CA, USA) using the paired-end (PE) technology within a single run, in which 100 bp PE reads were obtained. The original image process to sequences, base calling, and quality value calculation were performed using the Illumina GA Pipeline (version 1.6) [42,43]. All the sequencing processes of different B. sorokiniana samples were conducted in duplicates.…”

Section: Methodsmentioning

confidence: 99%

Disclosure of the Molecular Mechanism of Wheat Leaf Spot Disease Caused by Bipolaris sorokiniana through Comparative Transcriptome and Metabolomics Analysis

Liu

Zhang

et al. 2019

IJMS

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Wheat yield is greatly reduced because of the occurrence of leaf spot diseases. Bipolaris sorokiniana is the main pathogenic fungus in leaf spot disease. In this study, B. sorokiniana from wheat leaf (W-B. sorokiniana) showed much stronger pathogenicity toward wheat than endophytic B. sorokiniana from Pogostemon cablin (P-B. sorokiniana). The transcriptomes and metabolomics of the two B. sorokiniana strains and transcriptomes of B. sorokiniana-infected wheat leaves were comparatively analyzed. In addition, the expression levels of unigenes related to pathogenicity, toxicity, and cell wall degradation were predicted and validated by quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis. Results indicated that pathogenicity-related genes, especially the gene encoding loss-of-pathogenicity B (LopB) protein, cell wall-degrading enzymes (particularly glycosyl hydrolase-related genes), and killer and Ptr necrosis toxin-producing related unigenes in the W-B. sorokiniana played important roles in the pathogenicity of W-B. sorokiniana toward wheat. The down-regulation of cell wall protein, photosystem peptide, and rubisco protein suggested impairment of the phytosynthetic system and cell wall of B. sorokiniana-infected wheat. The up-regulation of hydrolase inhibitor, NAC (including NAM, ATAF1 and CUC2) transcriptional factor, and peroxidase in infected wheat tissues suggests their important roles in the defensive response of wheat to W-B. sorokiniana. This is the first report providing a comparison of the transcriptome and metabolome between the pathogenic and endophytic B. sorokiniana strains, thus providing a molecular clue for the pathogenic mechanism of W-B. sorokiniana toward wheat and wheat’s defensive response mechanism to W-B. sorokiniana. Our study could offer molecular clues for controlling the hazard of leaf spot and root rot diseases in wheat, thus improving wheat yield in the future.

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“…Moreover, LaeA [18] and VeA [19] global regulators have also been demonstrated to play an important role in the generation of gliotoxins in A. fumigutas. In addition, the in vitro function of genes related to gliotoxin biosynthesis, including gliZ, gliG, gliI, and gliO in deep-sea fungus Dichotomyces cejpii FS110 [20] and gliotoxin biosynthesis-related genes gliK, gliM, and gliT in deep-sea fungus Geosmithia pallida FS140 [21], have been characterized by their biochemical reaction. Interestingly, GliK was reported to exert the function of gliotoxin acetylation, thus facilitating the generation of gliotoxin derivates in G. pallida FS140.…”

Section: The Biosynthesis Of Gliotoxinmentioning

confidence: 99%

“…Gliotoxin and its derivatives have been excavated from deep-sea fungus D. cejpii FS110 [20] and G. pallida FS140 [21] in our group. Additionally, epipolythiodioxopiperazine derivatives of gliotoxin with the dithiol group showed weaker cytotoxicity compared with gliotoxin, and gliotoxin derivates of the methylthio group showed much weaker cytotoxicity than that of dithiol gliotoxin.…”

Section: The Biosynthesis Of Gliotoxinmentioning

confidence: 99%

The Toxic Mechanism of Gliotoxins and Biosynthetic Strategies for Toxicity Prevention

Liu

Zhang

2021

IJMS

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Gliotoxin is a kind of epipolythiodioxopiperazine derived from different fungi that is characterized by a disulfide bridge. Gliotoxins can be biosynthesized by a gli gene cluster and regulated by a positive GliZ regulator. Gliotoxins show cytotoxic effects via the suppression the function of macrophage immune function, inflammation, antiangiogenesis, DNA damage by ROS production, peroxide damage by the inhibition of various enzymes, and apoptosis through different signal pathways. In the other hand, gliotoxins can also be beneficial with different doses. Low doses of gliotoxin can be used as an antioxidant, in the diagnosis and treatment of HIV, and as an anti-tumor agent in the future. Gliotoxins have also been used in the control of plant pathogens, including Pythium ultimum and Sclerotinia sclerotiorum. Thus, it is important to elucidate the toxic mechanism of gliotoxins. The toxic mechanism of gliotoxins and biosynthetic strategies to reduce the toxicity of gliotoxins and their producing strains are summarized in this review.

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De Novo Transcriptome Sequencing of the Deep-Sea-Derived Fungus Dichotomomyces cejpii and Analysis of Gliotoxin Biosynthesis Genes

Cited by 6 publications

References 33 publications

Interaction of a Novel Zn2Cys6 Transcription Factor DcGliZ with Promoters in the Gliotoxin Biosynthetic Gene Cluster of the Deep-Sea-Derived Fungus Dichotomomyces cejpii

Interaction of a Novel Zn2Cys6 Transcription Factor DcGliZ with Promoters in the Gliotoxin Biosynthetic Gene Cluster of the Deep-Sea-Derived Fungus Dichotomomyces cejpii

Disclosure of the Molecular Mechanism of Wheat Leaf Spot Disease Caused by Bipolaris sorokiniana through Comparative Transcriptome and Metabolomics Analysis

The Toxic Mechanism of Gliotoxins and Biosynthetic Strategies for Toxicity Prevention

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