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Knockdown of arylalkylamine N ‐acetyltransferase‐like 2 in Drosophila melanogaster
Author(s) -
Anderson Ryan L.,
Wallis Dylan J.,
Aguirre Alexander,
Holliday Dean,
Merkler David J.
Publication year - 2019
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.21608
Subject(s) - drosophila melanogaster , biology , gene knockdown , melanogaster , transcription (linguistics) , gene expression , rna interference , microbiology and biotechnology , gene silencing , gene , biochemistry , rna , linguistics , philosophy
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.