The MSDIN family in amanitin-producing mushrooms and evolution of the prolyl oligopeptidase genes

Luo, Hong; Cai, Qing; Luli, Yungjiao; Li, Xuan; Sinha, Rohita; Hallen‐Adams, Heather E.; Yang, Zhu L.

doi:10.5598/imafungus.2018.09.02.01

Cited by 26 publications

(64 citation statements)

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“…It has been reported that HGT is very common in prokaryotes and can be an important source of their biological evolution, and HGT also occurs in eukaryotes at a lower frequency than that in prokaryotes [29][30][31][32]. The latest reports suggest that HGT may be responsible for the α-amanitin biosynthetic pathway found in the three distantly-related genera Amanita, Galerina and Lepiota [8,9]. It has been reported that in amanitin-producing mushrooms, the POPB gene catalyzes the cleavage and cyclization of the toxin precursor peptide, while the POPA gene is a housekeeping gene unrelated to toxin biosynthesis [7,12].…”

Section: Discussionmentioning

confidence: 99%

“…Recently, Pulman et al (2016) showed by generating draft genome sequences of A. palloides and A. bisporigera that both possessed approximately 30 members of the MSDIN family, but only three of these genes were common to both [6]. Luo et al (2018) found 18 and 22 MSDIN genes in the A. subjunquillea and A. pallidorosea genomes through PacBio and Illumina sequencing, respectively [8]. However, the MSDIN genes of many Amanita, Galerina and Lepiota amanitin-containing mushrooms have not been investigated in depth to date.…”

mentioning

confidence: 99%

“…According to previous research, whole-genome sequencing has proven to be the most comprehensive and in-depth method to identify MSDIN genes or genes related to the cyclopeptide biosynthetic pathway in amanitin-producing mushrooms [6,8]. Nevertheless, compared with genome sequencing, transcriptome sequencing provides an alternative efficient and low-cost method to obtain functional gene data.…”

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confidence: 99%

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Diversity of MSDIN family members in amanitin-producing mushrooms and the phylogeny of the MSDIN and prolyl oligopeptidase genes

Long

Fang

et al. 2020

Preprint

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Background Amanitin-producing mushrooms, mainly distributed in the genera Amanita , Galerina and Lepiota , possess MSDIN gene family for the biosynthesis of many cyclopeptides catalyzed by prolyl oligopeptidase (POP). Recently, transcriptome sequencing has proven to be a efficient way to mine MSDIN and POP genes in these lethal mushrooms. Until now, only A . palloides and A. bisporigera from North America and A . exitialis from Asia have been studied based on transcriptome analysis. However, MSDIN and POP genes of many amanitin-producing mushrooms in China remain unstudied, and hence the transcriptomes of these speices deserve to be analysed. Results In this study, the MSDIN and POP genes from ten Amanita species, two Galerina species and Lepiota venenata were studied and the phylogenetic relationships of their MSDIN and POP genes were analyzed. Through transcriptome sequencing and PCR cloning, 19 POP genes and 151 MSDIN genes predicted to encode 98 non-duplicated cyclopeptides, including α-amanitin, β-amanitin, phallacidin, phalloidin and 94 unknown peptides, were found in these species. Phylogenetic analysis showed that toxin peptide genes were clustered depending on the chemical substance within genus while depending on the taxonomy between genus and that the POPA genes of Amanita , Galerina and Lepiota were clustered and separated in three different groups, but the POPB genes of the three distinct genera were clustered in a highly monophyletic group. Conclusions These results above indicate that lethal Amanita species have the genetic capacity to produce numerous cyclopeptides, most of which are unknown, while lethal Galerina and Lepiota species seem to only have the genetic capacity to produce α-amanitin. Additionally, the POPB phylogeny of Amanita , Galerina and Lepiota conflicts with the taxonomic status of the three genera, suggesting that horizontal gene transfer might occur among the three genera.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Diversity of MSDIN family members in amanitin-producing mushrooms and the phylogeny of the MSDIN and prolyl oligopeptidase genes

Long

Fang

et al. 2020

Preprint

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“…unigenes. BLAST search against the transcriptome with MSDINs from the two As genomes produced 24 MSDINs, four more than those in our previous result [14]. These four sequences were used as queries to re-search the genome, and the result confirmed their presence ( In order to determine amino acid composition of CylG1 and CylG2, the candidate cyclic peptides were submitted to LC-MS/MS.…”

Section: Transcriptome Of Asmentioning

confidence: 94%

“…To achieve mature state of the toxins, many of these resultant cyclic peptides need to undergo further posttranslational modifications, mainly including multiple hydroxylations, sulfoxidation, epimerization, and formation of a cross-bridge between Trp and Cys, in which order these reactions occur is unknown. The MSDIN genes encoding amatoxins, such as α-and β-amanitins, and phallotoxins, such as phallacidin and phalloidin, are readily found in the sequenced genomes of lethal Amanita species [11,14]. In addition, there are 20 to 30 other unknown MSDIN genes discovered in each of these genomes.…”

Section: Introductionmentioning

confidence: 99%

Novel cyclic peptides from lethal Amanita species through a genomic approach, and major peptide toxins in the ectomycorrhizal association

Luo¹,

Zhou²,

Li³

et al. 2020

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Background: Most species in the genus Amanita are ectomycorrhizal fungi, and the cyclic peptide toxins that some species produce are notoriously deadly. In total, around 25 of these peptides were found in the fruiting bodies over the past 82 years, and whether any of them are present in the mycorrhizae is unknown. Reportedly, sequenced lethal Amanita genomes harbor a significant number of precursor genes of MSDIN family, indicating there could be a much larger capacity for cyclic peptide production in these mushrooms. However, it is largely unknown that to what extent these genes are transcribed, and further, translated into true cyclic peptides. Method:In this study, three poisonous Amanita species, A. rimosa, A. exitialis and A. subjunquillea, were sequenced through PacBio and Illumina techniques. For expression analysis, one strain of A.subjunquillea was sequenced through RNA-Seq. A genome-guided approach was adopted to identify cyclic peptides by coupling predicted toxin-biosynthetic genes with mass spectrometry (MS and MS/MS). To investigate whether any of the toxins were express in the microbiome, profiling of known major toxins was conducted on A. subjunquillea mycorrhizae via HRMS and gene cloning. Results:The resultant genomes showed significant potential to produce known and unknown cyclic peptides. Together with our 2 previously sequenced genomes, in total 37 unknown MSDIN genes were discovered. Expression of over 90% of the MSDIN genes was demonstrated in two strains of A. subjunquillea. Through the genome-guided approach, 12 MSDIN genes were found to produce true, novel cyclic peptides with no additional posttranslational modifications. When the ectomycorrhizae of A. subjunquillea were analyzed by MS, all major toxins were detected. The corresponding MSDINs for these cyclic peptides were successfully cloned directly from the mycorrhizae. Conclusions:The genome-guided approach provided a speedy method to identify cyclic peptides both in Amanita mushrooms and in the ectomycorrhizae. In this study, a significant number of novel MSDIN genes were discovered, most of which were found to be expressed in the tested species. The identification of the 12 novel cyclic peptides strongly suggests that Amanita species possess a much larger reservoir of these peptides than previously thought. This is the first report to demonstrate that the cyclic peptides in Amanita species are expressed in the mycorrhizal association. All four major

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Discovery of novel fungal RiPP biosynthetic pathways and their application for the development of peptide therapeutics

Vogt

Künzler

2019

Appl Microbiol Biotechnol

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Bioactive peptide natural products are an important source of therapeutics. Prominent examples are the 19 antibiotic penicillin and the immunosuppressant cyclosporine which are both produced by fungi and have 20 revolutionized modern medicine. Peptide biosynthesis can occur either non-ribosomally via large enzymes 21 referred to as non-ribosomal peptide synthetases (NRPS) or ribosomally. Ribosomal peptides are synthesized as 22 part of a larger precursor peptide where they are posttranslationally modified and subsequently proteolytically 23 released. Such peptide natural products are referred to as ribosomally synthesized and posttranslationallly 24 modified peptides (RiPPs). Their biosynthetic pathways have recently received a lot of attention, both from a 25 basic and applied research point of view, due to the discoveries of several novel posttranslational modifications 26 of the peptide backbone. Some of these modifications were so far only known from NRPSs and significantly 27 increase the chemical space covered by this class of peptide natural products. Latter feature, in combination with 28 the promiscuity of the modifying enzymes and the genetic encoding of the peptide sequence, makes RiPP 29 biosynthetic pathways attractive for synthetic biology approaches to identify novel peptide therapeutics via 30 screening of de novo generated peptide libraries and, thus, exploit bioactive peptide natural products beyond 31 their direct use as therapeutics. This review focuses on the recent discovery and characterization of novel RiPP 32 biosynthetic pathways in fungi and their possible application for the development of novel peptide therapeutics. 33 34 38 Meanwhile, the search for novel natural products with pharmacological activities continues. High-throughput 39 screens using molecular and cellular bioassays allow the evaluation of large numbers of natural products and 40 synthetically generated compound libraries for bioactivities of interest. Statistical analysis of the 41 complementarity of natural products and synthetic compounds showed that 40% of chemical scaffolds of natural 42 products are absent in synthetic compound libraries (Henkel et al. 1999), demonstrating that the biomedical 43 relevance of natural products lies in their wide range of structural diversity and complexity. Thus, natural 44 products continue to play a significant role in drug development. From the 1940s to 2014, 40% of small molecules 45 approved for cancer treatment were natural products or derivatives thereof (Newman and Cragg 2016). Plants 46 and microorganisms are the main sources of bioactive natural products (Dias et al. 2012). The vast dimension of 47 'microbial dark matter' suggested by genomic microbiome studies promises an enormous variety of bioactive48natural products yet to be discovered (Solden et al. 2016). This is also true for fungi where it is assumed that only 49 about one-twentieth of all existing fungi have been described and an even smaller fraction has been cultured 50 successfully in the lab (Jiang and An 2000...

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The MSDIN family in amanitin-producing mushrooms and evolution of the prolyl oligopeptidase genes

Cited by 26 publications

References 70 publications

Diversity of MSDIN family members in amanitin-producing mushrooms and the phylogeny of the MSDIN and prolyl oligopeptidase genes

Diversity of MSDIN family members in amanitin-producing mushrooms and the phylogeny of the MSDIN and prolyl oligopeptidase genes

Novel cyclic peptides from lethal Amanita species through a genomic approach, and major peptide toxins in the ectomycorrhizal association

Discovery of novel fungal RiPP biosynthetic pathways and their application for the development of peptide therapeutics

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