Study of DFF45 in its role of chaperone and inhibitor: two independent inhibitory domains of DFF40 nuclease activity.
CAD/DFF40, the nuclease responsible for DNA fragmentation during apoptosis, exists as a heterodimeric complex with DFF45/ICAD. This study determines the molecular mechanisms of regulation of DFF40 via the chaperone and inhibition activities of DFF45. We analyze proteins corresponding to the fragments (D1, D2, and D3) of DFF45 generated by cleavage at the caspase consensus sites in DFF45. Either D1 or D2, as an isolated domain, is capable of inhibiting DFF40 nuclease activity while double domain fragments D1-2 and D2-3, as well as full-length DFF45, bind to DFF40 with high affinity and are much more effective inhibitors. The chaperone activity of DFF45 resides in part in its ability to maintain DFF40 as a soluble protein. In addition, D1 of DFF45 was found to be critical for the expression of active DFF40 in vivo, suggesting a role for DFF45 in binding nascent DFF40.
McCarty
J S
JS
Laboratory of Apoptosis Regulation, Institute of Molecular and Cell Biology, 30 Medical Drive, Singapore, 117609, Singapore.
Toh
S Y
SY
Li
P
P
D051017:Apoptosis Regulatory Proteins; D020169:Caspases / Q000378:metabolism; D002478:Cells, Cultured; D003851:Deoxyribonucleases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D004791:Enzyme Inhibitors; D004794:Enzyme Repression; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D018832:Molecular Chaperones / Q000502:physiology*; D010446:Peptide Fragments / Q000378:metabolism; D000075223:Poly-ADP-Ribose Binding Proteins; D011506:Proteins / Q000502:physiology*
D051017:Apoptosis Regulatory Proteins; D004791:Enzyme Inhibitors; D047908:Intracellular Signaling Peptides and Proteins; D018832:Molecular Chaperones; D010446:Peptide Fragments; D000075223:Poly-ADP-Ribose Binding Proteins; D011506:Proteins; C109664:caspase-activated DNase inhibitor; C113696:DFFB protein, human; D003851:Deoxyribonucleases; D020169:Caspases