Genetic Variation of Adenylation Domains of the Anabaenopeptin Synthesis Operon and Evolution of Substrate Promiscuity

Christiansen, Guntram; Philmus, Benjamin; Hemscheidt, Thomas; Kurmayer, Rainer

doi:10.1128/jb.00360-11

Cited by 63 publications

(110 citation statements)

References 51 publications

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“…1 (25). In addition, smaller amounts of cyanopeptolins and aeruginosins can be found (25,28). The genetic manipulation of NIVA-CYA126/8 and the resulting mutants with knockout mutations for microcystins (NIVA-CYA126/8 McyϪ), anabaenopeptins (NIVA-CYA126/8 ApnϪ), and microviridin K (NIVA-CYA126/8 MvdϪ) were described earlier (14,15,25).…”

Section: Origin and Description Of Test Organisms A Detailed Descripmentioning

confidence: 99%

“…We used a simple but unique experimental setup that employed the bloom-forming cyanobacterium NIVA-CYA126/8 as a host model. NIVA-CYA126/8 is the first example of a cyanobacterium with a reasonably complex secondary metabolism for which knockout mutants for all of its major oligopeptide classes exist (14,15,25). These mutants differ from the wild type by their inability to produce microcystins, anabaenopeptins, or microviridins only, and so comparing the mutants with their wild type is one of the most direct ways to identify the significance of certain oligopeptide classes to biotic interactions.…”

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confidence: 99%

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Putative Antiparasite Defensive System Involving Ribosomal and Nonribosomal Oligopeptides in Cyanobacteria of the Genus Planktothrix

Rohrlack

Christiansen

Kurmayer

2013

Appl Environ Microbiol

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Parasitic chytrid fungi can inflict significant mortality on cyanobacteria but frequently fail to keep cyanobacterial dominance and bloom formation in check. Our study tested whether oligopeptide production, a common feature in many cyanobacteria, can be a defensive mechanism against chytrid parasitism. The study employed the cyanobacterial strain Planktothrix NIVA-CYA126/8 and its mutants with knockout mutations for microcystins, anabaenopeptins, and microviridins, major oligopeptide classes to be found in NIVA-CYA126/8. Four chytrid strains were used as parasite models. They are obligate parasites of Planktothrix and are unable to exploit alternative food sources. All chytrid strains were less virulent to the NIVA-CYA126/8 wild type than to at least one of its oligopeptide knockout mutants. One chytrid strain even failed to infect the wild type, while exhibiting considerable virulence to all mutants. It is therefore evident that producing microcystins, microviridins, and/or anabaenopeptins can reduce the virulence of chytrids to Planktothrix, thereby increasing the host's chance of survival. Microcystins and anabaenopeptins are nonribosomal oligopeptides, while microviridins are produced ribosomally, suggesting that Planktothrix resists chytrids by relying on metabolites that are produced via distinct biosynthetic pathways. Chytrids, on the other hand, can adapt to the oligopeptides produced by Planktothrix in different ways. This setting most likely results in an evolutionary arms race, which would probably lead to Planktothrix and chytrid population structures that closely resemble those actually found in nature. In summary, the findings of the present study suggest oligopeptide production in Planktothrix to be part of a defensive mechanism against chytrid parasitism.

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Section: Origin and Description Of Test Organisms A Detailed Descripmentioning

confidence: 99%

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confidence: 99%

Putative Antiparasite Defensive System Involving Ribosomal and Nonribosomal Oligopeptides in Cyanobacteria of the Genus Planktothrix

Rohrlack

Christiansen

Kurmayer

2013

Appl Environ Microbiol

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“…[6] To understand substrate selection, we determined its crystal structure and verified ligand binding by soaking the apo crystals in solutions containing several ligand compositions (Table S2 in the Supporting Information).…”

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Structural Elucidation of the Bispecificity of A Domains as a Basis for Activating Non‐natural Amino Acids

et al. 2015

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Many biologically active peptide secondary metabolites of bacteria are produced by modular enzyme complexes, the non-ribosomal peptide synthetases. Substrate selection occurs through an adenylation (A) domain, which activates the cognate amino acid with high fidelity. The recently discovered A domain of an Anabaenopeptin synthetase from Planktothrix agardhii (ApnA A1) is capable of activating two chemically distinct amino acids (Arg and Tyr). Crystal structures of the A domain reveal how both substrates fit into to binding pocket of the enzyme. Analysis of the binding pocket led to the identification of three residues that are critical for substrate recognition. Systematic mutagenesis of these residues created A domains that were monospecific, or changed the substrate specificity to tryptophan. The non-natural amino acid 4-azidophenylalanine is also efficiently activated by a mutant A domain, thus enabling the production of diversified non-ribosomal peptides for bioorthogonal labeling.

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“…It remains to be seen whether this is a feature of the remainder of the uncharacterized NRP synthetases within A. fumigatus and other fungi. Support for redundancy among NRP synthetases may come from observations that NRP synthetases show less strict substrate selection and incorporation than other adenylating enzymes (8,46).…”

Section: Discussionmentioning

confidence: 99%

Nonribosomal Peptide Synthetase Genes pesL and pes1 Are Essential for Fumigaclavine C Production in Aspergillus fumigatus

O’Hanlon

Gallagher

Schrettl

et al. 2012

Appl Environ Microbiol

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ABSTRACTThe identity of metabolites encoded by the majority of nonribosomal peptide synthetases in the opportunistic pathogen,Aspergillus fumigatus, remains outstanding. We found that the nonribosomal peptide (NRP) synthetases PesL and Pes1 were essential for fumigaclavine C biosynthesis, the end product of the complex ergot alkaloid (EA) pathway inA. fumigatus. Deletion of eitherpesL(ΔpesL) orpes1(Δpes1) resulted in complete loss of fumigaclavine C biosynthesis, relatively increased production of fumitremorgins such as TR-2, fumitremorgin C and verruculogen, increased sensitivity to H2O2, and increased sensitivity to the antifungals, voriconazole, and amphotericin B. Deletion ofpesLresulted in severely reduced virulence in an invertebrate infection model (P< 0.001). These findings indicate that NRP synthesis plays an essential role in mediating the final prenylation step of the EA pathway, despite the apparent absence of NRP synthetases in the proposed EA biosynthetic cluster forA. fumigatus. Liquid chromatography/diode array detection/mass spectrometry analysis also revealed the presence of fumiquinazolines A to F in bothA. fumigatuswild-type and ΔpesLstrains. This observation suggests that alternative NRP synthetases can also function in fumiquinazoline biosynthesis, since PesL has been shown to mediate fumiquinazoline biosynthesisin vitro. Furthermore, we provide here the first direct link between EA biosynthesis and virulence, in agreement with the observed toxicity associated with EA exposure. Finally, we demonstrate a possible cluster cross-talk phenomenon, a theme which is beginning to emerge in the literature.

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Genetic Variation of Adenylation Domains of the Anabaenopeptin Synthesis Operon and Evolution of Substrate Promiscuity

Cited by 63 publications

References 51 publications

Putative Antiparasite Defensive System Involving Ribosomal and Nonribosomal Oligopeptides in Cyanobacteria of the Genus Planktothrix

Putative Antiparasite Defensive System Involving Ribosomal and Nonribosomal Oligopeptides in Cyanobacteria of the Genus Planktothrix

Structural Elucidation of the Bispecificity of A Domains as a Basis for Activating Non‐natural Amino Acids

Nonribosomal Peptide Synthetase Genes pesL and pes1 Are Essential for Fumigaclavine C Production in Aspergillus fumigatus

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