Molecular characterization of the lysosomal acid phosphatase from Drosophila melanogaster.
In Drosophila, unlike humans, the lysosomal acid phosphatase (Acph-1) is a non-essential enzyme. It is also one of the most rapidly evolving gene-enzyme systems in the genus. In order to determine which parts of the enzyme are conserved and which parts are apparently under little functional constraint, we cloned the gene from Drosophila melanogaster via a chromosomal walk. Fragments from the gene were used to recover an apparently full-length cDNA. The cDNA was sub-cloned into a Drosophila transformation vector where it was under the control of the 5' promoter sequence of the hsp-70 gene. Three independent transformants were obtained; in each, Acph-1 expression from the cDNA was constitutive and not dependent on heat shock, as determined by densitometric analyses of the allozymic forms of the enzyme. The pattern of expression indicates the hsp-70 and endogenous Acph-1 promoters act together in some, but not all, tissues. The sequence of the cDNA was determined using deletions made with exonuclease III, and primers deduced from the cDNA sequence were used to sequence the genomic clone. Five introns were found, and putative 5' upstream regulatory sequences were identified. Amino acid sequence comparisons have revealed several highly conserved motifs between Drosophila Acph-1 and vertebrate lysosomal and prostatic acid phosphatases.
Chung
H J
HJ
Section of Genetics and Development, Cornell University, Ithaca, New York 14853, USA.
Shaffer
C
C
MacIntyre
R
R
D000135:Acid Phosphatase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016386:Chromosome Walking; D003001:Cloning, Molecular; D016384:Consensus Sequence; D017124:Conserved Sequence; D018076:DNA, Complementary; D004331:Drosophila melanogaster / Q000201:enzymology* / Q000235:genetics; D015870:Gene Expression; D017344:Genes, Insect; D006801:Humans; D007438:Introns; D008247:Lysosomes / Q000201:enzymology*; D008969:Molecular Sequence Data; D010802:Phylogeny; D011994:Recombinant Proteins / Q000737:chemistry; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D011994:Recombinant Proteins; D000135:Acid Phosphatase