Molecular and Biochemical Characterization of the aaNATl (Dat) Locus in Drosophila melanogaster: Differential Expression of Two Gene Products

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Abstract
In insects, arylalkylamine N-acetyltransferases (AANATs) have been implicated in several physiological processes, including sclerotization, inactivation of certain neurotransmitters, and, similar to the function in vertebrates, catalysis of the rate-limiting step in melatonin biosynthesis. Here, we report an extensive biochemical and functional analysis of the products of the aaNAT1 gene of Drosophila melanogaster. The aaNAT1 gene generates two transcripts through alternative first-exon usage. These transcripts are under tissue-specific and developmental control and encode proteins which differ in their N-terminus with respect to their starting methionine. The more abundant isoform, AANAT1b, is first expressed during late embryogenesis in the brain, the ventral nerve cord, and the midgut; in adults, AANAT1b is still detectable in the brain and midgut. The less abundant isoform, AANAT1a, appears only during late pupal stages and in adults is found predominantly in the brain. We demonstrate that the mutation Datlo represents a hypomorphic allele of aaNAT1b, in which an insertion of two transposable elements, MDG412 and blastopia, has occurred within the first intron of the gene. Using a deficiency which removes the aaNAT1 gene, we provide evidence that aaNAT1 is not essential for the process of sclerotization. Furthermore, neither of the two enzyme isoforms shows circadian regulation of RNA or protein levels. The differing levels of abundance and distinct developmental control of AANAT1a and AANAT1b suggest different in vivo functions for these two enzymes.