Abstract
Ergot alkaloids are fungal natural products with important roles in agriculture and medicine. We used heterologous expression and gene knockout approaches to investigate potential roles for the product of a major facilitator superfamily transporter gene (easT) recently found in an ergot alkaloid biosynthetic gene cluster in Aspergillus leporis. A strain of Aspergillus fumigatus previously engineered to accumulate lysergic acid, but which did not convert the precursor agroclavine to lysergic acid efficiently or secrete lysergic acid well, was chosen as an expression host for easT. Expression of easT in this strain resulted in accumulation of significantly more pathway intermediates but no detectable lysergic acid. Secretion of ergot alkaloids was reduced in the easT-expressing strain. EasT localized to discrete vesicle-like structures in the cytosol of A. fumigatus, with no localization detected in the plasma membrane. When easT was knocked out in A. leporis, accumulation of lysergic acid amides was reduced relative to the wild type. There was no negative effect on secretion of ergot alkaloids in the knockout mutant. The data indicate that easT encodes a product that contributes to accumulation of ergot alkaloids, perhaps by transporting intermediates between cellular compartments, but does not have a significant role in secreting ergot alkaloids.
Original language | English |
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Pages (from-to) | 406-417 |
Number of pages | 12 |
Journal | Applied Microbiology |
Volume | 4 |
Issue number | 1 |
DOIs | |
State | Published - Mar 2024 |
Externally published | Yes |
Funding
This research was funded by the National Institute of General Medical Sciences, grant number R15-GM114774, with additional salary support to D.G.P. from USDA NIFA Hatch funds. A.M.J. is a Beckman Scholar supported by the Arnold and Mabel Beckman Foundation. Imaging experiments were performed at the West Virginia University (WVU) Microscope Imaging Facility which has been supported by the WVU Cancer Institute, the WVU HSC Office of Research and Graduate Education, and NIH grants P20GM121322 and P20GM144230.
Keywords
- Aspergillus
- ergot alkaloids
- lysergic acid
- lysergic acid amides
- major facilitator superfamily
- transporter