A single locus encodes both phenylalanine hydroxylase and tryptophan hydroxylase activities in Drosophila.
We have used a full-length clone encoding rabbit tryptophan hydroxylase (TRH) to isolate the Drosophila homologue (DTPH). Southern analysis of Drosophila genomic DNA reveals a pattern indicative of a single gene. The single transcript is expressed in adult head and body mRNA but is also detected in mRNA from early embryos. The embryonic transcript is ubiquitously expressed and appears to concentrate in yolk granules. In situ hybridization of TRH-homologous antisense RNA probe to sectioned tissue from third instar larvae demonstrated the presence of this transcript in fat body and cuticular tissue. Developmental immunoblot analysis using antibodies raised against a beta-galactosidase-Drosophila fusion protein revealed a 45-kDa embryonic protein also detected in female abdomens and a 50-kDa protein found in larval and adult stages. Immunocytochemical analysis of the Drosophila protein in the larval central nervous system showed that it appeared to be present in both serotonin- and catecholamine-containing neurons. A nonfusion protein generated in Escherichia coli hydroxylates both tryptophan and phenylalanine. We propose that there are only two aromatic amino acid hydroxylase genes in Drosophila: one encoding tyrosine hydroxylase, DTH, and DTPH, a gene encoding both tryptophan and phenylalanine hydroxylase activities.
Department of Biology, Brandeis University, Waltham, Massachusetts 02254.
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Neckameyer
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White
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002874:Chromosome Mapping; D004247:DNA / Q000235:genetics; D004331:Drosophila melanogaster / Q000201:enzymology / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D015870:Gene Expression; D007150:Immunohistochemistry; D008297:Male; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010651:Phenylalanine Hydroxylase / Q000235:genetics* / Q000378:metabolism; D012313:RNA / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015183:Restriction Mapping; D014365:Tryptophan Hydroxylase / Q000235:genetics* / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D012313:RNA; D004247:DNA; D010651:Phenylalanine Hydroxylase; D014365:Tryptophan Hydroxylase; D001616:beta-Galactosidase