Pyrin N-terminal homology domain- and caspase recruitment domain-dependent oligomerization of ASC.
ASC was first identified as a caspase recruitment domain (CARD)-containing proapoptotic molecule that forms insoluble aggregates during apoptosis. Here, we report both the pyrin N-terminal homology domain (PYD) and CARD domains are involved in the aggregation of ASC. Preliminary experiments indicated that overexpression of ASC formed filament-like aggregates in COS-7 cells. Expression experiments using green fluorescent protein (GFP) constructs showed that not only the GFP-ASC-CARD but also the GFP-ASC-PYD formed filament-like aggregates in COS-7 cells. We confirmed these filament-like aggregates of both the ASC-PYD and the ASC-CARD due to homophilic interaction by immunoprecipitation method. We also demonstrated that the ASC-PYD associated with the ASC-CARD by heterophilic interaction. These observations suggest that the dimerization of the PYD as well as the CARD plays an important role in the oligomerization of ASC as an adaptor molecule.
Masumoto
J
J
Department of Molecular Oncology and Angiology, Research Center on Aging and Adaptation, Shinshu University School of Medicine, Asahi 3-1-1, Matsumoto 390-8621, Nagano, Japan.
Taniguchi
S
S
Sagara
J
J
D000818:Animals; D053468:CARD Signaling Adaptor Proteins; D019556:COS Cells; D020169:Caspases / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019281:Dimerization; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry*; D000071198:Pyrin; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D053468:CARD Signaling Adaptor Proteins; D003598:Cytoskeletal Proteins; D008164:Luminescent Proteins; C000604192:MEFV protein, human; C402202:PYCARD protein, human; D011506:Proteins; D000071198:Pyrin; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D020169:Caspases