Knockdown of arylalkylamine N-acetyltransferase-like 2 in Drosophila melanogaster
Autor: | Dean Holliday, David J. Merkler, Ryan L. Anderson, Dylan J. Wallis, Alexander Aguirre |
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Rok vydání: | 2019 |
Předmět: |
0106 biological sciences
0301 basic medicine Physiology AANAT Dopamine 01 natural sciences Biochemistry Article 03 medical and health sciences Gene expression Melanogaster Animals Drosophila Proteins Gene Silencing Gene knockdown biology General Medicine biology.organism_classification Cell biology 010602 entomology RNA silencing 030104 developmental biology Real-time polymerase chain reaction Drosophila melanogaster Insect Science Arylalkylamine Acyltransferases |
Zdroj: | Archives of insect biochemistry and physiology. 102(4) |
ISSN: | 1520-6327 |
Popis: | 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. |
Databáze: | OpenAIRE |
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