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Journal of Neuroscience, Vol 10, 1929-1942, Copyright © 1990 by Society for Neuroscience
Interspecific comparison of a Drosophila gene encoding FMRFamide- related neuropeptides
PH Taghert and LE Schneider
Department of Anatomy and Neurobiology, Washington University School of Medicine, Saint Louis, Missouri 63110.
In order to identify functionally important regions of a neuropeptide gene
in Drosophila melanogaster, we have studied its occurrence in related
species and have characterized the structure of a homologous gene in
Drosophila virilis. The melanogaster gene encodes a precursor that contains
13 neuropeptides related to the molluscan tetrapeptide FMRFamide (Nambu et
al., 1988; Schneider and Taghert, 1988). Using the melanogaster gene as a
probe in Southern blot analysis, related sequences were detected in DNA
from each of 7 species tested. D. virilis, which is estimated to have
diverged from D. melanogaster between 60 and 80 million years ago
(Throckmorton, 1975), was chosen for more detailed study.
Immunocytochemical staining using an antibody to authentic FMRFamide
revealed a similar set of immunoreactive neurons in the CNS of larvae from
the 2 Drosophila species. Using a melanogaster gene probe, overlapping
clones were isolated from a virilis genomic library; DNA sequence analysis
indicated the presence of a homologous gene. Comparisons of the genes and
deduced proteins between the 2 species revealed the following points. (1)
Both genes are divided into 2 exons: in D. melanogaster the exons are 106
and 1352 bp long; in D. virilis, they are 169 and at least 1232 bp long; in
both species, the intron is approximately 2.5 kb long. (2) The sequence of
exon I has largely diverged, and in neither species are exon I sequences
translated. In this vicinity of the gene, sequence conservation is limited
to a 67 bp region that spans the TATA box and the RNA start site. (3) The
deduced neuropeptide precursors have very similar sizes (347 vs 339 amino
acids) and the presumed signal sequences are perfectly conserved. (4) While
the melanogaster precursor contains 13 FMRFamide-related peptides, the
virilis precursor contains only 10. (5) The sequences of some but not all
of the FMRFamide-like peptides are perfectly conserved. (6) In the rest of
the precursor, significant sequence conservation is found only in the
N-terminal portion; immediately downstream of the final FMRFamide-like
peptide, the protein sequences are highly divergent. (7) 5' to the RNA
start sites (1.2 kb of melanogaster DNA and 1.8 kb of virilis DNA), 17
small (9-52 base pairs) regions are evolutionarily conserved (greater than
80% sequence conservation). We discuss neuropeptide biosynthesis, the
functions and evolution of FMRFamide-like neuropeptides in insects, and the
cell-specific regulation of neuropeptide gene expression in the contexts of
these results.
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