The fungal gene cluster for biosynthesis of the antibacterial agent viriditoxin

Urquhart, Andrew S.; Hu, Jinyu; Chooi, Yit‐Heng; Idnurm, Alexander

doi:10.1101/601716

Cited by 5 publications

(11 citation statements)

References 50 publications

(52 reference statements)

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“…For pcaA the regions of homology were amplified using primer pairs AP219 and AP220 and AP223 and AP224. The GFP - hph region was amplified from a GFP-tagging construct generated previously 67 using primers AP221 and AP222. For zrcA the regions of homology were amplified using primer pairs AP299 and AP302 and AP305 and AP307.…”

Section: Methodsmentioning

confidence: 99%

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Eukaryotic transposable elements as “cargo carriers”: the forging of metal resistance in the fungusPaecilomyces variotii

Urquhart

Chong

Yang

et al. 2020

Preprint

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29The horizontal transfer of large gene clusters by mobile elements is a key driver of prokaryotic 30 adaptation in response to environmental stresses. Eukaryotic microbes face similar 31 environmental stresses yet a parallel role for mobile elements has not yet been established. A 32 stress faced by all microorganisms is the prevalence of toxic metals in their environment. In 33 fungi, identified mechanisms for protection against metals generally rely on genes that are 34 dispersed within an organism's genome. Here we have discovered a large (~85 kb) region that 35 confers resistance to several metals in the genomes of some, but not all, strains of a fungus, 36Paecilomyces variotii. We name this region HEPHAESTUS (Hϕ) and present evidence that this 37 region is mobile within the P. variotii genome with features highly characteristic of a 38 transposable element. While large gene clusters including those for the synthesis of secondary 39 metabolites have been widely reported in fungi, these are not mobile within fungal genomes. 40 HEPHAESTUS contains the greatest complement of host-beneficial genes carried by a 41 transposable element in eukaryotes. This suggests that eukaryotic transposable elements might 42 play a role analogous to their bacterial counterparts in the horizontal transfer of large regions of 43 host-beneficial DNA. Genes within HEPHAESTUS responsible for individual metal resistances 44 include those encoding a P-type ATPase transporter, PcaA, required for cadmium and lead 45 resistance, a transporter, ZrcA, providing resistance to zinc, and a multicopper oxidase, McoA, 46 conferring resistance to copper. Additionally, a subregion of Hϕ conferring resistance to 47 arsenate was identified. The presence of a strikingly similar cluster in the genome of another 48 fungus, Penicillium fuscoglaucum, suggests that HEPHAESTUS arrived in P. variotii via horizontal 49 gene transfer.50 51 KEYWORDS 52 53 Eurotiales, gene cluster, metal homeostasis, horizontal gene transfer, transposon 54 3 INTRODUCTION 55 56Metals are used within diverse fields such as agriculture, construction, electronics and 57 pharmaceutical science. A number of these, such as cadmium, lead and arsenic, are toxic even 58 at low levels of exposure and thus environmental contamination resulting from their use is an 59 increasing concern 1 . The prevalence of toxic metals in the environment is of particular 60 relevance to fungal biology. Some fungi are able to accumulate metals to high concentrations, 61 making such fungi dangerous for human consumption 2,3 , allowing bioremediation of 62 contaminated sites 4 or concentrating valuable metals for extraction 5 . Zinc and copper play 63 essential roles in many cellular functions and are often important in fungal virulence 6 . However, 64 at high concentrations these metals can also be toxic; for example, copper-based molecules are 65 frequently used as fungicides in agriculture. 66 67 Compared to prokaryotes, our understanding of the evolutionary mechanisms through which 68 eukaryotes have ada...

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Section: Methodsmentioning

confidence: 99%

“…The RNA was reverse transcribed using AMV reverse transcriptase (New England Biolabs) using oligo-dT. The mcoA gene was amplified using primers AP369 and AP370 and the constitutive beta-tubulin gene was amplified as described previously 67 .…”

Section: Methodsmentioning

confidence: 99%

Eukaryotic transposable elements as “cargo carriers”: the forging of metal resistance in the fungusPaecilomyces variotii

Urquhart

Chong

Yang

et al. 2020

Preprint

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“…For pcaA the regions of homology used for targeted integration into the native locus on a 5' flank and 3' flank were amplified using primer pairs AP219-AP220 and AP223-AP224. The GFP-hph region was amplified from a GFP-tagging construct generated previously 90 using primers AP221-AP222. For zrcA the regions of homology were amplified using primer pairs AP299-AP302 and AP305-AP307.…”

Section: Cloning and Dna Manipulationsmentioning

confidence: 99%

“…The mcoA gene was amplified using primers AP369-AP370, and the constitutive beta-tubulin gene was amplified with primers b-tubu-linqPCRF and b-tubulinqPCRR as designed previously. 90…”

Section: Supplemental Informationmentioning

confidence: 99%

A large transposable element mediates metal resistance in the fungus Paecilomyces variotii

et al. 2022

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A large transposable element mediates metal resistance in the fungus Paecilomyces variotiiHighlights d A gene cluster in the fungus Paecilomyces variotii confers metal resistance d The gene cluster displays hallmarks of coordinated movement as a mobile element d The cluster is variably located in different genomic locations in different strains d HEPHAESTUS is an example of transposons carrying beneficial traits in eukaryotes Authors

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“…In many ascomycetes, disruption of a specific polyketide synthase (PKS) gene results in loss of conidial pigmentation. As a consequence, these transformants produce lighter or white conidia [46][47][48][49]. Comparing the conidia of these mutants with their parental strain will lead to new insights into the potential roles of melanin in preservation stress resistance of conidia.…”

Section: Fungal Biology and Biotechnologymentioning

confidence: 99%

Preservation stress resistance of melanin deficient conidia from Paecilomyces variotii and Penicillium roqueforti mutants generated via CRISPR/Cas9 genome editing

Seekles

Teunisse

Punt

et al. 2021

Fungal Biol Biotechnol

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Background The filamentous fungi Paecilomyces variotii and Penicillium roqueforti are prevalent food spoilers and are of interest as potential future cell factories. A functional CRISPR/Cas9 genome editing system would be beneficial for biotechnological advances as well as future (genetic) research in P. variotii and P. roqueforti. Results Here we describe the successful implementation of an efficient AMA1-based CRISPR/Cas9 genome editing system developed for Aspergillus niger in P. variotii and P. roqueforti in order to create melanin deficient strains. Additionally, kusA− mutant strains with a disrupted non-homologous end-joining repair mechanism were created to further optimize and facilitate efficient genome editing in these species. The effect of melanin on the resistance of conidia against the food preservation stressors heat and UV-C radiation was assessed by comparing wild-type and melanin deficient mutant conidia. Conclusions Our findings show the successful use of CRISPR/Cas9 genome editing and its high efficiency in P. variotii and P. roqueforti in both wild-type strains as well as kusA− mutant background strains. Additionally, we observed that melanin deficient conidia of three food spoiling fungi were not altered in their heat resistance. However, melanin deficient conidia had increased sensitivity towards UV-C radiation.

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The fungal gene cluster for biosynthesis of the antibacterial agent viriditoxin

Cited by 5 publications

References 50 publications

Eukaryotic transposable elements as “cargo carriers”: the forging of metal resistance in the fungusPaecilomyces variotii

Eukaryotic transposable elements as “cargo carriers”: the forging of metal resistance in the fungusPaecilomyces variotii

A large transposable element mediates metal resistance in the fungus Paecilomyces variotii

Preservation stress resistance of melanin deficient conidia from Paecilomyces variotii and Penicillium roqueforti mutants generated via CRISPR/Cas9 genome editing

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