Knockdown of Arylalkylamine N-acetyltransferase-like 2 in Drosophila Melanogaster
Document Type
Article
Publication Date
2019
Keywords
arylalkylamine N-acetyltransferase, Drosophila melanogaster, expression knockdown, fatty acid amide, LC-QToF-MS, N-palmitoyldopamine, UAS/Gal4
Digital Object Identifier (DOI)
https://doi.org/10.1002/arch.21608
Abstract
Drosophila melanogaster produces fatty acid amides, and thus, provides a model to unravel the pathways for their biosynthesis. We previously demonstrated that arylalkylamine N-acetyltransferase-like 2 (AANATL2) from D. melanogaster will catalyze the formation of long-chain N-acylserotonins and N-acyldopamines in vitro. Generating silencing RNA via the UAS/GAL4 bipartite approach for targeted gene expression effectively decreased the endogenous levels of the AANATL2 transcripts in D. melanogaster, as shown by reverse transcription quantitative polymerase chain reaction. Consistent with these data, western blot analysis of the offspring of the AANATL2 knockdown flies using an anti-AANATL2 antibody revealed a significant reduction in the expression of the AANATL2 protein. Reduced expression of AANATL2 decreased the cellular levels of N-palmitoyldopamine (PALDA), providing strong evidence that AANATL2 is responsible for the biosynthesis of PALDA in vivo. This is the first time that the expression of an AANAT has been reduced in D. melanogaster to link one of these enzymes to the in vivo production of an N-acylarylalkylamide.
Was this content written or created while at USF?
Yes
Citation / Publisher Attribution
Archives of Insect Biochemistry and Physiology, v. 102, issue 4, art. e21608
Scholar Commons Citation
Anderson, Ryan L.; Wallis, Dylan J.; Aguirre, Alexander; Holliday, Dean; and Merkler, David J., "Knockdown of Arylalkylamine N-acetyltransferase-like 2 in Drosophila Melanogaster" (2019). Chemistry Faculty Publications. 140.
https://digitalcommons.usf.edu/chm_facpub/140
