Detection of poly(ADP-ribose) synthesis in Drosophila testes upon gamma-irradiation.
Poly(ADP-ribose) polymerase (PARP) activity is widespread among eukaryotes. Upon DNA damage PARP binds to DNA strand breaks and transfers ADP-ribose residues from NAD+ to acceptor proteins and to ADP-ribosyl protein adducts. This leads to branched polymers of protein-coupled poly(ADP-ribose) (pADPr). Because the germline of Drosophila has recently become important in the study of DNA double-strand break repair (DSBR) as opposed to somatic DSBR we tested whether the catalytic activity of PARP can be stimulated by gamma-irradiation during Drosophila spermatogenesis. Using antibodies against pADPr we detected a significant increase in PARP activity in male germline cells during spermatogenesis upon gamma-irradiation. Different stages of spermatogenesis revealed different subnuclear localization patterns of pADPr. In premeiotic and postmeiotic cells pADPr localized in a pattern overlapping with lamin and topoisomerase II at the nuclear rim. In primary spermatocytes pADPr is associated with three loci corresponding to the chromosomes at the nuclear periphery.
Lankenau
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Department of Tumor Virology, Deutsches Krebsforschungszentrum Heidelberg, Im Neuenheimer Feld 280, D-69120 Heidelberg, Germany.
Bürkle
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Lankenau
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DH
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism / Q000528:radiation effects; D004249:DNA Damage; D004250:DNA Topoisomerases, Type II / Q000378:metabolism; D004307:Dose-Response Relationship, Radiation; D004331:Drosophila melanogaster / Q000378:metabolism* / Q000528:radiation effects*; D004789:Enzyme Activation / Q000528:radiation effects; D007150:Immunohistochemistry; D034882:Lamins; D008297:Male; D009687:Nuclear Proteins / Q000378:metabolism; D011064:Poly Adenosine Diphosphate Ribose / Q000096:biosynthesis*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D013091:Spermatogenesis / Q000502:physiology / Q000528:radiation effects; D013737:Testis / Q000378:metabolism* / Q000528:radiation effects*
D034882:Lamins; D009687:Nuclear Proteins; D011064:Poly Adenosine Diphosphate Ribose; D011065:Poly(ADP-ribose) Polymerases; D004250:DNA Topoisomerases, Type II