Biosynthetic Pathway for the Epipolythiodioxopiperazine Acetylaranotin in Aspergillus terreus Revealed by Genome-Based Deletion Analysis

Guo, Chun-Jun; Yeh, Hsu-Hua; Chiang, Yi‐Ming; Sanchez, James F.; Chang, Shu-Ling; Bruno, Kenneth S.; Wang, Clay C. C.

doi:10.1021/ja3123653

Cited by 85 publications

(130 citation statements)

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“…18 In addition, all data for the ring parts in 6 were similar to those of known bisdethiobis(methylthio)-deacetylapoaranotin (10). 15 The cross-peaks from 8-H (δ 5.88) to C-11 (δ 171.5) and from 8′-H (δ 6.22) to C-11′ (δ 171.8) in HMBC data supported the same acyl side chains was attached at both C-8 and C-8′. Taken together, the structure of 6 was elucidated as shown in Figure 1.…”

Section: )mentioning

confidence: 73%

“…We started to search for such inhibitors from natural sources using our established screening system, 12 leading to the discovery of tylopilusins 13 and citridone A and its derivatives. 14 Continuing our screening program, new compounds designated graphiumins A (1) to H (8), along with two structurally related known bisdethiobis(methylthio)-deacetylaranotin (9) 15 and bisdethiobis(methylthio)-deacetylapoaranotin (10), 15 were isolated from the culture broth of the marine-derived fungus Graphium sp. OPMF00224 (Figure 1).…”

Section: Introductionmentioning

confidence: 99%

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Graphiumins, new thiodiketopiperazines from the marine-derived fungus Graphium sp. OPMF00224

et al. 2015

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Eight new thiodiketopiperazines, designated as graphiumins A to H (1-8), were isolated along with bisdethiobis(methylthio)-deacetylaranotin (9) and bisdethiobis(methylthio)-deacetylapoaranotin (10) from the culture broth of the marine-derived fungus Graphium sp. OPMF00224. The structures of the graphiumins were elucidated based on spectroscopic analyses (1D and 2D NMR data, ROESY correlations and CD data) and chemical methods. The absolute configuration of the common (3S)-3-hydroxy-octanoyl acid residue in 1, 3 and 4 was determined by hydrolysis, benzoyl derivatization and HPLC analysis using a chiral column. Five graphiumins moderately inhibited yellow pigment production by methicillin-resistant Staphylococcus aureus.

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Section: )mentioning

confidence: 73%

Section: Introductionmentioning

confidence: 99%

Graphiumins, new thiodiketopiperazines from the marine-derived fungus Graphium sp. OPMF00224

et al. 2015

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“…HPLC-MS was carried out using a ThermoFinnigan LCQ Advantage ion-trap mass spectrometer with a reverse-phase C18 column (Alltech Prevail C18; 2.1 mm × 100 mm with a 3-μm particle size) at a flow rate of 125 μL/min. The solvent-gradient system for HPLC and the conditions for MS analysis were carried out as described previously (47).…”

Section: Methodsmentioning

confidence: 99%

Prototype of an intertwined secondary-metabolite supercluster

Wiemann

Guo²,

Palmer

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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The hallmark trait of fungal secondary-metabolite gene clusters is well established, consisting of contiguous enzymatic and often regulatory gene(s) devoted to the production of a metabolite of a specific chemical class. Unexpectedly, we have found a deviation from this motif in a subtelomeric region of Aspergillus fumigatus. This region, under the control of the master regulator of secondary metabolism, LaeA, contains, in its entirety, the genetic machinery for three natural products (fumitremorgin, fumagillin, and pseurotin), where genes for fumagillin and pseurotin are physically intertwined in a single supercluster. Deletions of 29 adjoining genes revealed that fumagillin and pseurotin are coregulated by the supercluster-embedded regulatory gene with biosynthetic genes belonging to one of the two metabolic pathways in a noncontiguous manner. Comparative genomics indicates the fumagillin/pseurotin supercluster is maintained in a rapidly evolving region of diverse fungal genomes. This blended design confounds predictions from established secondary-metabolite cluster search algorithms and provides an expanded view of natural product evolution.gene regulation | Zn(II) 2 Cys 6 transcription factor | FapR | biosynthesis | cluster evolution F ilamentous fungi are well known for their ability to produce a variety of natural products, so-called secondary metabolites that are not essential for growth under laboratory conditions (reviewed in refs. 1 and 2). However, the maintenance of the genetic information allowing fungi to produce secondary metabolites suggests that these small molecules provide essential benefits in environmental niches ranging from protection from fungivory (reviewed in ref.3) to chemical shields from UV radiation (4). Apart from providing evolutionary fitness to the producing organism in their natural habitat, many secondary metabolites are of major importance to humans because of their beneficial and deleterious effects as drugs and toxins, respectively.Fungal secondary metabolism has been characterized by physical linkage of the genes required for synthesis of specific metabolites and the distinct enzymatic machinery encoded by these genes (1, 2, 5). For instance, most fungal secondary metabolites belong to one of four chemical classes that are characterized based on the key or backbone enzymes that consist of polyketide synthases (PKSs), nonribosomal peptide synthetases (NRPSs), terpene cyclases (TCs), and prenyltransferases (PTs). Typically, cluster genes adjacent to these backbone genes code for accessory enzymes involved in either modification of the chemical scaffold, transcriptional control of cluster genes, transport of substrates and/or products, and resistance mechanisms. The most common regulatory genes of clusters encode fungal-specific C6 zinc binuclear cluster (Zn(II) 2 Cys 6 ) transcription factors (6), which, in general, exert positive transcriptional regulation of most of the genes within a single cluster (7). In addition to cluster-specific transcription factors, a higher order...

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“…iosynthesis, self-protection mechanisms, and functionality of gliotoxin and related epidithiodiketopiperazine (ETP) molecular species, such as chaetocin and acetylaranotin, are attracting ever-increasing attention as a consequence of findings from highthroughput genome sequencing projects, application of gene deletion technologies, and mass spectrometric analytical methodologies (1)(2)(3)(4)(5). Indeed, existing paradigms of gliotoxin ( Fig.…”

mentioning

confidence: 99%

“…Deletion of hlmI impaired holomycin biosynthesis and sensitized S. clavuligerus to exogenous holomycin, as had been observed for gliotoxin in A. fumigatus. Additionally, Guo et al revealed that in Aspergillus terreus, a fusion gene, ataTC, encodes both hydroxylation and disulfide bridge closure of biosynthetic intermediates during acetylaranotin formation (3). Wild-type A. terreus did not exhibit acetylaranotin sensitivity; however, no data were presented regarding the sensitivity of the ⌬ataTC strain to exogenous acetylaranotin.…”

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confidence: 99%

Interplay between Gliotoxin Resistance, Secretion, and the Methyl/Methionine Cycle in Aspergillus fumigatus

et al. 2015

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Mechanistic studies on gliotoxin biosynthesis and self-protection in Aspergillus fumigatus, both of which require the gliotoxin oxidoreductase GliT, have revealed a rich landscape of highly novel biochemistries, yet key aspects of this complex molecular architecture remain obscure. Here we show that an A. fumigatus ⌬gliA strain is completely deficient in gliotoxin secretion but still retains the ability to efflux bisdethiobis(methylthio)gliotoxin (BmGT). This correlates with a significant increase in sensitivity to exogenous gliotoxin because gliotoxin trapped inside the cell leads to (i) activation of the gli cluster, as disabling gli cluster activation, via gliZ deletion, attenuates the sensitivity of an A. fumigatus ⌬gliT strain to gliotoxin, thus implicating cluster activation as a factor in gliotoxin sensitivity, and (ii) increased methylation activity due to excess substrate (dithiol gliotoxin) for the gliotoxin bis-thiomethyltransferase GtmA. Intracellular dithiol gliotoxin is oxidized by GliT and subsequently effluxed by GliA. In the absence of GliA, gliotoxin persists in the cell and is converted to BmGT, with levels significantly higher than those in the wild type. Similarly, in the ⌬gliT strain, gliotoxin oxidation is impeded, and methylation occurs unchecked, leading to significant S-adenosylmethionine (SAM) depletion and S-adenosylhomocysteine (SAH) overproduction. This in turn significantly contributes to the observed hypersensitivity of gliT-deficient A. fumigatus to gliotoxin. Our observations reveal a key role for GliT in preventing dysregulation of the methyl/methionine cycle to control intracellular SAM and SAH homeostasis during gliotoxin biosynthesis and exposure. Moreover, we reveal attenuated GliT abundance in the A. fumigatus ⌬gliK strain, but not the ⌬gliG strain, following exposure to gliotoxin, correlating with relative sensitivities. Overall, we illuminate new systems interactions that have evolved in gliotoxin-producing, compared to gliotoxin-naive, fungi to facilitate their cellular presence. Biosynthesis, self-protection mechanisms, and functionality of gliotoxin and related epidithiodiketopiperazine (ETP) molecular species, such as chaetocin and acetylaranotin, are attracting ever-increasing attention as a consequence of findings from highthroughput genome sequencing projects, application of gene deletion technologies, and mass spectrometric analytical methodologies (1-5). Indeed, existing paradigms of gliotoxin (Fig. 1) as a toxin and the perspective of the disulfide bridge-containing (oxidized) form as the final, or only, product are undergoing significant reconsideration (6-11).Self-protection against disulfide-containing metabolites appears to be essential in both fungi and bacteria. It has been demonstrated that the gliotoxin oxidoreductase GliT (12), encoded within the gli cluster, protects Aspergillus fumigatus against exogenous gliotoxin and is essential for gliotoxin biosynthesis (12, 13). A similar mechanism for self-protection against holomycin in Streptomyces clavulige...

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Biosynthetic Pathway for the Epipolythiodioxopiperazine Acetylaranotin in Aspergillus terreus Revealed by Genome-Based Deletion Analysis

Cited by 85 publications

References 41 publications

Graphiumins, new thiodiketopiperazines from the marine-derived fungus Graphium sp. OPMF00224

Graphiumins, new thiodiketopiperazines from the marine-derived fungus Graphium sp. OPMF00224

Prototype of an intertwined secondary-metabolite supercluster

Interplay between Gliotoxin Resistance, Secretion, and the Methyl/Methionine Cycle in Aspergillus fumigatus

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