10.5061/DRYAD.0RXWDBS0T
Panaccione, Daniel
0000-0002-4159-164X
West Virginia University
Steen, Chey
West Virginia University
Chemical analyses and insect interactions of an easO mutant of Metarhizium
brunneum
Dryad
dataset
2021
FOS: Biological sciences
National Cancer Institute
https://ror.org/040gcmg81
2R15-GM114774-2
2021-06-23T00:00:00Z
2021-06-23T00:00:00Z
en
https://doi.org/10.1128/AEM.00748-21
69001 bytes
7
CC0 1.0 Universal (CC0 1.0) Public Domain Dedication
Several fungi, including the plant root symbiont and insect pathogen
Metarhizium brunneum, produce lysergic acid amides via a branch of the
ergot alkaloid pathway. Lysergic acid amides include important
pharmaceuticals and pharmaceutical lead compounds and have potential
ecological significance, making knowledge of their biosynthesis relevant.
Many steps in the biosynthesis of lysergic acid amides have been
determined, but terminal steps in the synthesis of lysergic acid
α-hydroxyethylamide (LAH)––by far the most abundant lysergic acid amide in
M. brunneum––are unknown. Ergot alkaloid synthesis (eas) genes are
clustered in the genomes of fungi that produce these compounds, and the
eas clusters of LAH producers contain two uncharacterized genes (easO and
easP) not found in fungi that do not produce LAH. Knockout of easO via a
CRISPR-Cas9 approach eliminated LAH and resulted in accumulation of
alternate lysergic acid amides lysergyl-alanine and ergonovine. Despite
the elimination of LAH, the total concentration of lysergic acid
derivatives was not affected significantly by the mutation.
Complementation with a wild-type allele of easO restored the ability to
synthesize LAH. Substrate feeding studies indicated that neither
lysergyl-alanine nor ergonovine were substrates for the product of easO
(EasO). EasO had structural similarity to Baeyer-Villiger monooxygenases
(BVMOs), and labeling studies with deuterated alanine supported a role for
a BVMO in LAH biosynthesis. The easO knockout had reduced virulence to
larvae of the insect Galleria mellonella, indicating that LAH contributes
to virulence of M. brunneum on insects and that LAH has biological
activities different from ergonovine and lysergyl-alanine.
Data were collected as described in Steen et al. 2021. Applied and
Environmental Microbiology 88:e00748-21.
Data are contained in an excel file with separate tabs for each of the
major data set. The tab labeled “hplc data” contains data contributing to
Table 1 of the Applied and Environmental Microbiology article. The “deu
ala” tab contains data supporting the statistical analyses in Fig. 3. The
tab labeled “insect survival” contains insect survival data used to
prepare Figure 4. The “spores” tab contains data used to prepare Figure
5. The data in each tab are arrayed to be copied and pasted into a JMP
worksheet.