Mechanism of Human SIRT1 Activation by Resveratrol*

By M. Borra, Brian C. Smith, J. Denu. Journal of Biological Chemistry, 2005

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The NAD+-dependent protein deacetylase family, Sir2 (or sirtuins), is important for many cellular processes including gene silencing, regulation of p53, fatty acid metabolism, cell cycle regulation, and life span extension. Resveratrol, a polyphenol found in wines and thought to harbor major health benefits, was reported to be an activator of Sir2 enzymes in vivo and in vitro. In addition, resveratrol was shown to increase life span in three model organisms through a Sir2-dependent pathway. Here, we investigated the molecular basis for Sir2 activation by resveratrol. Among the three enzymes tested (yeast Sir2, human SIRT1, and human SIRT2), only SIRT1 exhibited significant enzyme activation (∼8-fold) using the commercially available Fluor de Lys kit (BioMol). To examine the requirements for resveratrol activation of SIRT1, we synthesized three p53 acetylpeptide substrates either lacking a fluorophore or containing a 7-amino-4-methylcoumarin (p53-AMC) or rhodamine 110 (p53-R110). Although SIRT1 activation was independent of the acetylpeptide sequence, resveratrol activation was completely dependent on the presence of a covalently attached fluorophore. Substrate competition studies indicated that the fluorophore decreased the binding affinity of the peptide, and, in the presence of resveratrol, fluorophore-containing substrates bound more tightly to SIRT1. Using available crystal structures, a model of SIRT1 bound to p53-AMC peptide was constructed. Without resveratrol, the coumarin of p53-AMC peptide is solvent-exposed and makes no significant contacts with SIRT1. We propose that binding of resveratrol to SIRT1 promotes a conformational change that better accommodates the attached coumarin group.

Subfields: Epidemiology, Geriatrics and Gerontology, Pathology and Forensic Medicine

View Paper: https://www.semanticscholar.org/paper/2fff24819752a5d1c4f643e5e71399d9542f0a87

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Citations in the corpus

Child health, developmental plasticity, and epigenetic programming. (2011, background): “For example, resveratrol, a polyphenol constituent of red wine, can activate human SIRT1, which is member of the family of sirtuin proteins that are essential for gene silencing (541) (see Section II.”
Therapeutic potential of resveratrol: the in vivo evidence (2006, background): “The in vitro activity of SIRT1 against short, unconjugated peptide substrates is not enhanced by resveratrol, leading some to argue that the original observation was an artefact of the fluorescent substrate used to monitor the reactio”
AMP-Activated Protein Kinase–Deficient Mice Are Resistant to the Metabolic Effects of Resveratrol (2009, background): “However, whether the putative Sirt1 activators such as resveratrol actually target Sirt1 in vivo is controversial because resveratrol increases Sirt1 activity in vitro only if the substrate is modified with a fluorescent tag (9,10).”
Resveratrol Ameliorates Aging-Related Metabolic Phenotypes by Inhibiting cAMP Phosphodiesterases (2012, neutral): “DISCUSSION By demonstrating that resveratrol activates the cAMP-Epac1-AMPK-Sirt1 pathway, this study, in conjunction with previous studies (Beher et al., 2009; Borra et al., 2005; Kaeberlein et al., 2005; Pacholec et al., 2010), explains how resveratrol activates Sirt1 without directly targeting it.”
SIRT1 is required for AMPK activation and the beneficial effects of resveratrol on mitochondrial function (2012, background): “One school of thought is that the direct activation of SIRT1 by resveratrol is an in vitro artifact (Beher et al., 2009; Borra et al., 2005; Kaeberlein et al., 2005; Pacholec et al., 2010) and that resveratrol works primarily by activating AMPK (Canto et al.”
Age Related Changes in NAD+ Metabolism Oxidative Stress and Sirt1 Activity in Wistar Rats (2011, background): “While several studies have demonstrated the protective roles of sirtuin activators during aging [30,34,35,58,59,60,61,62,63], little is known regarding role of Sirt1 in the aging heart, lung, liver, and kidney.”
Resveratrol improves mitochondrial function and protects against metabolic disease by activating SIRT1 and PGC-1alpha. (2006, methodology): “Since such a convergence could potentially impact on metabolic diseases, we addressed our hypothesis not in the context of CR but under conditions of caloric excess using the specific SIRT1 activator, resveratrol (RSV) (Borra et al., 2005; Howitz et al., 2003).”
Sirtuins in aging and age-related disease. (2006, background): “However, other groups have shown that the stimulatory effects of resveratrol on Sirtuin enzyme activation are specific to this nonnative substrate (Borra et al., 2005; Kaeberlein et al., 2005c).”
Resveratrol prolongs lifespan and retards the onset of age-related markers in a short-lived vertebrate. (2006, background): “The effect of resveratrol is mimicked by overexpression of sirtuins [6, 7, 28], but does not seem to be mediated by direct action on sirtuins [32, 33].”
Molecular mechanisms and potential therapeutical targets in Huntington's disease. (2010, background): “Interestingly, activation, rather than inhibition of sirtuins, with the small molecule resveratrol was found neuroprotective in HD worms (73, 431).”