The enzymatic component of Drosophila melanogaster chorion is the Pxd peroxidase.
In the present study, we demonstrate the isolation and characterization of the Pxd cDNA clone, which codes for the Drosophila melanogaster chorion peroxidase. This specific peroxidase is involved in the chorion hardening process, through protein crosslinking mediated by the formation of di- and tri-tyrosine bonds. The Pxd gene product has been identified in crude protein extracts from adult flies as three immunoreacting, with the anti-rAePO polyclonal antibody, bands of 77, 67 and 55 kDa, while in larvae and purified chorions as a unique 55 kDa band. Moreover, the mature form of the Pxd recombinant protein was specifically recognized by the anti-rAePO antibody as a 77 kDa band, while in the presence of H2O2 was able to convert tyrosine residues to di-tyrosine moieties. Northern blotting analysis of total RNA preparations revealed distinct molecular weight patterns of the Pxd RNA transcripts among adult flies, ovaries and larvae. The in situ hybridization clearly shows that the Pxd mRNA is specifically expressed in follicle cells during the late stages of oogenesis 11-14, while the reverse transcription reactions dictate the stage-specific developmental regulation of the Pxd gene. The immunolocalization approach, using the anti-rAePO polyclonal antibody, has revealed that the Pxd peroxidase is selectively localized in the chorion structures and particularly in the endochorion and innermost chorionic layer (ICL).
Department of Cell Biology and Biophysics, Faculty of Biology, University of Athens, Panepistimiopolis, 15784 Athens, Greece.
Ourania A
OA
Konstandi
Issidora S
IS
Papassideri
Dimitrios J
DJ
Stravopodis
Christos A
CA
Kenoutis
Zulfiqar
Z
Hasan
Theodoros
T
Katsorchis
Ron
R
Wever
Lukas H
LH
Margaritis
D000818:Animals; D002823:Chorion / Q000201:enzymology; D004331:Drosophila melanogaster / Q000201:enzymology*; D005786:Gene Expression Regulation; D019476:Insect Proteins / Q000737:chemistry* / Q000378:metabolism; D010544:Peroxidases / Q000737:chemistry* / Q000378:metabolism
D019476:Insect Proteins; D010544:Peroxidases