SMAC negatively regulates the anti-apoptotic activity of melanoma inhibitor of apoptosis (ML-IAP).
Inhibitors of apoptosis (IAPs) physically interact with a variety of pro-apoptotic proteins and inhibit apoptosis induced by diverse stimuli. X-linked IAP (X-IAP) is a prototype IAP family member that inhibits several caspases, the effector proteases of apoptosis. The inhibitory activity of X-IAP is regulated by SMAC, a protein that is processed to its active form upon receipt of a death stimulus. Cleaved SMAC binds X-IAP and antagonizes its anti-apoptotic activity. Here we show that melanoma IAP (ML-IAP), a potent anti-cell death protein and caspase inhibitor, physically interacts with SMAC through its BIR (baculovirus IAP repeat) domain. In addition to binding full-length SMAC, ML-IAP BIR associates with SMAC peptides that are derived from the amino terminus of active, processed SMAC. This high affinity interaction is very specific and can be completely abolished by single amino acid mutations either in the amino terminus of active SMAC or in the BIR domain of ML-IAP. In cells expressing ML-IAP and X-IAP, SMAC coexpression or addition of SMAC peptides abrogates the ability of the IAPs to inhibit cell death. These results demonstrate the feasibility of using SMAC peptides as a way to sensitize IAP-expressing cells to pro-apoptotic stimuli such as chemotherapeutic agents.
Vucic
Domagoj
D
Department of Molecular Oncology, Genentech, Inc., South San Francisco, California 94080, USA.
Deshayes
Kurt
K
Ackerly
Heidi
H
Pisabarro
Maria Teresa
MT
Kadkhodayan
Saloumeh
S
Fairbrother
Wayne J
WJ
Dixit
Vishva M
VM
D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D017209:Apoptosis; D002352:Carrier Proteins / Q000378:metabolism*; D053453:Caspase 9; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D015938:Complement Membrane Attack Complex; D003165:Complement System Proteins; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D051034:Inhibitor of Apoptosis Proteins; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D009363:Neoplasm Proteins / Q000378:metabolism*; D009707:Nucleosomes / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D013997:Time Factors; D014162:Transfection; D014407:Tumor Cells, Cultured
D048868:Adaptor Proteins, Signal Transducing; C419866:BIRC7 protein, human; D002352:Carrier Proteins; D015938:Complement Membrane Attack Complex; D006023:Glycoproteins; D051034:Inhibitor of Apoptosis Proteins; D009363:Neoplasm Proteins; D009707:Nucleosomes; D010455:Peptides; C047610:SC5b-9 protein complex; D003165:Complement System Proteins; C506433:CASP9 protein, human; D053453:Caspase 9; D020169:Caspases