Sex-peptides bind to two molecularly different targets in Drosophila melanogaster females.
Sex-Peptide (SP) and the peptide DUP99B elicit two postmating responses in Drosophila melanogaster females: receptivity is reduced and oviposition is increased. Both are synthesized in the male genital tract and transferred into the female during copulation. To elucidate their function, we characterized the binding properties of SP and DUP99B in females. Cryostat sections of adult females were incubated with alkaline phosphatase (AP)-tagged peptides. In virgin females, both peptides have specific target sites in the nervous system and in the genital tract. The binding pattern is almost identical for both peptides. Incubation of sections of mated females confirm that some of these target sites correspond to the in vivo targets of the two peptides. Neuronal binding is dependent on an intact C-terminal sequence of SP, binding in the genital tract is less demanding in terms of amino acid sequence requirement. On affinity blots the AP-SP probe binds to membrane proteins extracted from abdomen and head plus thorax, respectively. The binding proteins in the nervous system and the genital tract differ in their molecular properties. Calculation of dissociation constants (K(d)), and also determination of the minimal peptide concentrations necessary for binding, indicate that SP is the more important peptide inducing the postmating responses. Our results suggest that binding of SP in the nervous system is responsible for eliciting the postmating responses, whereas binding in the genital tract reflects the presence of a peptide transporter.
Ding
Zhaobing
Z
Zoologisches Institut Universität Zürich-Irchel, Winterthurerstrasse190, CH-8057 Zürich, Switzerland.
Haussmann
Irmgard
I
Ottiger
Michael
M
Kubli
Eric
E
D000818:Animals; D029721:Drosophila Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000502:physiology*; D005260:Female; D005836:Genitalia, Female / Q000378:metabolism; D015151:Immunoblotting; D036341:Intercellular Signaling Peptides and Proteins; D009420:Nervous System / Q000378:metabolism; D010455:Peptides / Q000378:metabolism*; D011485:Protein Binding; D012724:Sex Attractants / Q000378:metabolism*; D012726:Sexual Behavior, Animal / Q000502:physiology*
D029721:Drosophila Proteins; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; D012724:Sex Attractants; C489177:Acp36DE protein, Drosophila; C449520:ductus ejaculatorius peptide 99B, Drosophila; C056736:SP protein, Drosophila