Expression and Functional Characterization of aDrosophila Neuropeptide Precursor with Homology to Mammalian Preprotachykinin A

doi:10.1074/jbc.M002875200

Expression and Functional Characterization of aDrosophila Neuropeptide Precursor with Homology to Mammalian Preprotachykinin A*

From the ^‡School of Biology, University of Leeds, Leeds, West Yorkshire LS2 9JT, United Kingdom, the ^**Department of Biological Sciences, University of Lancaster, Lancaster LA1 4YQ, United Kingdom, the ^¶Department of Zoology, Stockholm University, S-10691 Stockholm, Sweden, the ^‖Southern Plains Agricultural Research Center, United States Department of Agriculture, College Station, Texas 77845, and the ^§Department of Biology, Birkbeck College, University of London, London WC1E 7HX, United Kingdom

Abstract

Peptides structurally related to mammalian tachykinins have recently been isolated from the brain and intestine of several insect species, where they are believed to function as both neuromodulators and hormones. Further evidence for the signaling role of insect tachykinin-related peptides was provided by the cloning and characterization of cDNAs for two tachykinin receptors fromDrosophila melanogaster. However, no endogenous ligand has been isolated for the Drosophila tachykinin receptors to date. Analysis of the Drosophila genome allowed us to identify a putative tachykinin-related peptide prohormone (prepro-DTK) gene. A 1.5-kilobase pair cDNA amplified from aDrosophila head cDNA library contained an 870-base pair open reading frame, which encodes five novelD rosophila tachykinin-related peptides (called DTK peptides) with conserved C-terminal FXGXR-amide motifs common to other insect tachykinin-related peptides. The tachykinin-related peptide prohormone gene (Dtk) is both expressed and post-translationally processed in larval and adult midgut endocrine cells and in the central nervous system, with midgut expression starting at stage 17 of embryogenesis. The predictedDrosophila tachykinin peptides have potent stimulatory effects on the contractions of insect gut. These data provide additional evidence for the conservation of both the structure and function of the tachykinin peptides in the brain and gut during the course of evolution.

Footnotes

↵* This work was funded by Biotechnology and Biological Sciences Research Council Grants 24/S09564 and 89/S09563 (to R. E. I., D. C., and A. D. S.), by a grant from the Swedish Natural Science Research Council (to D. R. N.), and by NATO Grant 973325 (to G. M. C. and R. J. N.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) .
↵‡ To whom correspondence should be addressed. Tel.: 44-1132-332890; Fax: 44-1132-332835; E-mail: r.e.isaac@leeds.ac.uk.
Published, JBC Papers in Press, May 8, 2000, DOI 10.1074/jbc.M002875200
↵2 R. J. Siviter, unpublished results.
Abbreviations:

PPT-A

preprotachykinin A

TRP

tachykinin-related peptide

DTK

Drosophila tachykinin-related peptide

BAC

bacterial artificial chromosome

RACE

rapid amplification of cDNA ends

bp

base pair(s)

LemTRP

Le ucophaea m aderae tachykinin-related peptide

LomTk-II

Locusta migratoria tachykinin-II

TAP

tachykinin-associated peptide

LTK-LI

L ocusta tachykinin-likeimmunoreactive
- Received April 5, 2000.
The American Society for Biochemistry and Molecular Biology, Inc.