Sirt1 activation by resveratrol is substrate sequence-selective

Abstract

Sirtuins are protein deacetylases used as therapeutic targets. Pharmacological Sirt1 activation has been questioned since the in vitro activator resveratrol failed to stimulate deacetylation of several physiological substrates. We tested the influence of substrate sequence by analyzing resveratrol effects on Sirt1-dependent deacetylation of 6802 physiological acetylation sites using peptide microarrays. Resveratrol stimulated deacetylation of a small set of sites and inhibited deacetylation of another set, whereas most substrates were hardly affected. Solution assays confirmed these substrate categories, and statistical analysis revealed their sequence features. Our results reveal substrate sequence dependence for Sirt1 modulation and suggest substrates contributing to resveratrol effects.One Sentence SummaryTesting 6802 acetylation sites reveals that resveratrol effects on Sirt1-dependent deacetylation depend on substrate sequence and suggests substrates relevant for in vivo effects.

Resveratrol enhanced FOXO3 phosphorylation via synergetic activation of SIRT1 and PI3K/Akt signaling to improve the effects of exercise in elderly rat hearts

“…Resveratrol is also a SIRT1 activator (Frazzi et al 2013;Lakshminarasimhan et al 2013;Li et al 2013). When SIRT1 is activated, SIRT1 will block FOXO3 accumulation in the nucleus and inhibit cell death.…”

“…SIRT1 is a deacetylase that can remove the acetyl group from FOXO3 and NFκB (Frazzi et al 2013;Lakshminarasimhan et al 2013;Li et al 2013). …”

Ex-527 inhibits Sirtuins by exploiting their unique NAD + -dependent deacetylation mechanism

“…Recombinant human Sirt1, Sirt3, and Sirt5 were purified as described previously (12,45). Briefly, His-tagged full-length Sirt1 (in pET15b), His-Trxtagged Sirt3 residues 118-399 (in pVFT3S), and His-tagged Sirt5 residues 34-302 (in pET151/D-TOPO) were expressed in Escherichia coli BL21DE3 Rosetta2 (Novagen) and purified by affinity chromatography with Talon resin (Clontech).…”

Structural basis for allosteric, substrate-dependent stimulation of SIRT1 activity by resveratrol

“…However, several studies questioned the effectiveness of resveratrol and related compounds as direct SIRT1 activators, pointing out that the observed stimulatory effect depended on the presence of a fluorescent group in the substrates used in in vitro assays (Borra et al 2005;Kaeberlein et al 2005;Pacholec et al 2010). Nevertheless, it is possible that the artificial fluorescent group in the substrate peptide may mimic an aromatic residue in certain natural substrates of SIRT1 (Dai et al 2010;Hubbard et al 2013;Lakshminarasimhan et al 2013;Sinclair and Guarente 2014).…”