Inhibition of mono-ADP-ribosyltransferase activity during the execution phase of apoptosis prevents apoptotic body formation.
The objective of this study was to understand factors responsible for apoptotic body formation and release during apoptosis. We have found that inhibition of mono-ADP ribosylation after ultraviolet (UV) light induction of apoptosis in HL-60 cells does not block caspase-3 activation, gelsolin cleavage, or endonucleolytic DNA fragmentation. However, the cytoskeletal features of apoptosis leading to apoptotic body formation and release were inhibited by meta-iodobenzylguanidine (MIBG) and novobiocin, potent inhibitors of arginine-specific mono-ADP-ribosyltransferases (mono-ADPRTs). Suppression of mono-ADP ribosylation as late as 120 min following UV irradiation blocked the depolymerization of actin and release of apoptotic bodies. This suggested that the cytoskeletal changes of apoptosis may be decoupled from the caspase cascade and that there may be a biochemical event either distal to or independent of caspase-3 that regulates apoptotic body formation. To test the hypothesis that ADP ribosylation of actin may occur with the induction of apoptosis, an in vivo assay of mono-ADPRT activity using an antibody against ADP-ribosylarginine was used. An approximately 64% increase in the ADP ribosylation of actin was observed at 2 h following exposure to UV light. When MIBG or novobiocin was present, the ADP ribosylation of actin was only 14-18% above the levels observed in control nonirradiated cells. The current study is the first to demonstrate a relationship between ADP-ribosylation of actin and the formation of apoptotic bodies.
Lodhi
I J
IJ
Department of Surgery, Veterans Affairs Medical Center, University of Michigan Medical School, Ann Arbor 48105, USA.
Clift
R E
RE
Omann
G M
GM
Sweeney
J F
JF
McMahon
K K
KK
Hinshaw
D B
DB
D019797:3-Iodobenzylguanidine / Q000494:pharmacology; D036002:ADP Ribose Transferases / Q000037:antagonists & inhibitors*; D000199:Actins / Q000032:analysis; D017209:Apoptosis / Q000502:physiology*; D053148:Caspase 3; D020169:Caspases; D004720:Endonucleases / Q000378:metabolism; D018260:Gelsolin / Q000378:metabolism; D018922:HL-60 Cells / Q000528:radiation effects; D006801:Humans; D009675:Novobiocin / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D014466:Ultraviolet Rays
D000199:Actins; D018260:Gelsolin; D009675:Novobiocin; D019797:3-Iodobenzylguanidine; D036002:ADP Ribose Transferases; D004720:Endonucleases; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases