PARP cleavage in the apoptotic pathway in S2 cells from Drosophila melanogaster.
Caspase activities and two cDNA sequences have been identified in Drosophila melanogaster. To study the molecular events following the activation of the apoptotic pathway in D. melanogaster, S2 cells were treated with etoposide and the timing of the apoptotic events, such as caspase activation, mitochondrial pore opening, and loss of membrane asymmetry, was determined. Poly(ADP-ribose) polymerase (PARP) is known to be cleaved in the early phase of apoptosis in vertebrate systems. Little is known about the involvement of PARP cleavage in apoptosis in invertebrates. If PARP inactivation is a general event, this could mean that DNA repair enzymes need to be cleaved for the death pathway to be completed. We have found that in etoposide-treated cells, PARP protein is processed, but the nature of the cleavage is not known. Further experiments must be conducted and the peptide fragments must be sequenced to relate protease activities with PARP cleavage.
Poltronieri
P
P
Department of Biochemistry, Institute of Basic Medical Sciences, Tennoudai, Tsukuba, Japan. miro@md.tsukuba.ac.jp
Yokota
T
T
Koyama
Y
Y
Hanai
S
S
Uchida
K
K
Miwa
M
M
D000818:Animals; D017302:Annexins / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D003546:Cysteine Endopeptidases / Q000378:metabolism; D004331:Drosophila melanogaster / Q000166:cytology / Q000201:enzymology*; D004789:Enzyme Activation / Q000187:drug effects; D005047:Etoposide / Q000494:pharmacology; D016650:Fluorescein-5-isothiocyanate; D006868:Hydrolysis; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D011419:Propidium / Q000378:metabolism; D012235:Rhodamines / Q000378:metabolism
D017302:Annexins; D012235:Rhodamines; D011419:Propidium; D005047:Etoposide; D011065:Poly(ADP-ribose) Polymerases; D003546:Cysteine Endopeptidases; D016650:Fluorescein-5-isothiocyanate