Interaction with Sug1 enables Ipaf ubiquitination leading to caspase 8 activation and cell death

Kumar, Yatender ; Radha, Vegesna ; Swarup, Ghanshyam (2010) Interaction with Sug1 enables Ipaf ubiquitination leading to caspase 8 activation and cell death Biochemical Journal, 427 . pp. 91-104. ISSN 0006-2936

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Activation of initiator caspases is dependent on interacting proteins, and Ipaf [ICE (interleukin-1β -converting enzyme)-protease activating factor] {NLRC4 [NLR (Nod-like receptor) family CARD (caspase activation and recruitment domain)-containing 4]} an inflammasome component, is involved in caspase 1 activation and apoptosis. Investigating the mechanisms of Ipaf activation, we found that the C-terminal LRR (leucine-rich repeat) domain of Ipaf, through intramolecular interaction, negatively regulates its apoptosis-inducing function. In A549 lung carcinoma cells, expression of Ac-Ipaf (LRR-domain-deleted Ipaf) induced cell death that was dependent on caspase 8, but not on caspase 1. A yeast two-hybrid screen using Ac-Ipaf as bait identified human Sug1 (suppressor of gal 1), a component of the 26S proteasome, as an interacting protein. In mammalian cells Sug1 interacts and co-localizes with Ipaf. Sug1 binds to amino acids 91-253 of Ipaf, which is also the region that the LRR domain binds to. It potentiates cell death induced by Ipaf and Ac-Ipaf, and co-expression of Sug1 and Ipaf induces caspase-8-dependent cell death. Cellular complexes formed by Ipaf and Sug1 contain caspase 8. Expression of Ac-Ipaf or co-expression of Sug1 with Ipaf results in the formation of cytoplasmic aggregates and caspase 8 activation. Sug1 co-expression enabled modification of Ipaf by ubiquitination. Tagging ubiquitin molecules to Ipaf led to aggregate formation, enhanced caspase 8 interaction and activation, resulting in induction of cell death. Using RNAi (RNA interference) and dominant-negative approaches, we have shown that cell death induced by Ac-Ipaf expression or by treatment with TNF-α (tumour necrosis factor α ) or doxorubicin is dependent on Sug1. Our results suggest a role for ubiquitination of Ipaf that is enabled by its interaction with Sug1, leading to caspase 8 activation and cell death.

Item Type:Article
Source:Copyright of this article belongs to Biochemical Society.
Keywords:Aggresome; Caspase 8; Interleukin-1β-converting Enzyme-protease-activating Factor(Ipaf); Leucine-rich Repeat Domain (LRR Domain); Suppressor of Gal 1(Sug1); Ubiquitination
ID Code:55930
Deposited On:19 Aug 2011 07:57
Last Modified:19 Aug 2011 07:57

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