SIRT1 ubiquitination is regulated by opposing activities of APC/C-Cdh1 and AROS during stress-induced premature senescence

Abstract

SIRT1, a member of the mammalian sirtuin family, is a nicotinamide adenosine dinucleotide (NAD)-dependent deacetylase with key roles in aging-related diseases and cellular senescence. However, the mechanism by which SIRT1 protein homeostasis is controlled under senescent conditions remains elusive. Here, we revealed that SIRT1 protein is significantly downregulated due to ubiquitin-mediated proteasomal degradation during stress-induced premature senescence (SIPS) and that SIRT1 physically associates with anaphase-promoting complex/cyclosome (APC/C), a multisubunit E3 ubiquitin ligase. Ubiquitin-dependent SIRT1 degradation is stimulated by the APC/C coactivator Cdh1 and not by the coactivator Cdc20. We found that Cdh1 depletion impaired the SIPS-promoted downregulation of SIRT1 expression and reduced cellular senescence, likely through SIRT1-driven p53 inactivation. In contrast, AROS, a SIRT1 activator, reversed the SIRT1 degradation induced by diverse stressors and antagonized Cdh1 function through competitive interactions with SIRT1. Furthermore, our data indicate opposite roles for Cdh1 and AROS in the epigenetic regulation of the senescence-associated secretory phenotype genes IL-6 and IL-8. Finally, we demonstrated that pinosylvin restores downregulated AROS (and SIRT1) expression levels in bleomycin-induced mouse pulmonary senescent tissue while repressing bleomycin-promoted Cdh1 expression. Overall, our study provides the first evidence of the reciprocal regulation of SIRT1 stability by APC/C-Cdh1 and AROS during stress-induced premature senescence, and our findings suggest pinosylvin as a potential senolytic agent for pulmonary fibrosis. Aging: protecting a protein that promotes proliferationTwo opposing molecular mechanisms determine the stability of SIRT1, a protein that prevents cells from prematurely entering the 'retirement' state known as senescence. This state, in which cell division is halted and tissue repair and regeneration impeded, is a natural endpoint of aging but also arises from cellular damage and other stressors. South Korean researchers led by Eun-Joo Kim at Dankook University, Cheonan, and Soo-Jong Um at Sejong University, Seoul, have identified two proteins that govern the onset of senescence by affecting SIRT1's stability. One promotes the addition of chemical modifications to SIRT1 that mark it for destruction, while the other prevents such modifications. The researchers further showed that a drug that tips the balance in favor of SIRT1 stabilization could prevent early senescence in an animal model of pulmonary fibrosis, a currently incurable lung disorder.