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A human tRNA synthetase is a potent PARP1-activating effector target for resveratrol.

Sajish M, Schimmel P - Nature (2014)

Bottom Line: Resveratrol is reported to extend lifespan and provide cardio-neuro-protective, anti-diabetic, and anti-cancer effects by initiating a stress response that induces survival genes.Because human tyrosyl transfer-RNA (tRNA) synthetase (TyrRS) translocates to the nucleus under stress conditions, we considered the possibility that the tyrosine-like phenolic ring of resveratrol might fit into the active site pocket to effect a nuclear role.In contrast to functionally diverse tRNA synthetase catalytic s created by alternative splicing events that ablate active sites, here a non-spliced TyrRS catalytic reveals a new PARP1- and NAD(+)-dependent dimension to the physiological mechanism of resveratrol.

View Article: PubMed Central - PubMed

Affiliation: The Skaggs Institute for Chemical Biology, The Scripps Laboratories for tRNA Synthetase Research, Department of Molecular and Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.

ABSTRACT
Resveratrol is reported to extend lifespan and provide cardio-neuro-protective, anti-diabetic, and anti-cancer effects by initiating a stress response that induces survival genes. Because human tyrosyl transfer-RNA (tRNA) synthetase (TyrRS) translocates to the nucleus under stress conditions, we considered the possibility that the tyrosine-like phenolic ring of resveratrol might fit into the active site pocket to effect a nuclear role. Here we present a 2.1 Å co-crystal structure of resveratrol bound to the active site of TyrRS. Resveratrol ifies the catalytic activity and redirects TyrRS to a nuclear function, stimulating NAD(+)-dependent auto-poly-ADP-ribosylation of poly(ADP-ribose) polymerase 1 (PARP1). Downstream activation of key stress signalling pathways are causally connected to TyrRS-PARP1-NAD(+) collaboration. This collaboration is also demonstrated in the mouse, and is specifically blocked in vivo by a resveratrol-displacing tyrosyl adenylate analogue. In contrast to functionally diverse tRNA synthetase catalytic s created by alternative splicing events that ablate active sites, here a non-spliced TyrRS catalytic reveals a new PARP1- and NAD(+)-dependent dimension to the physiological mechanism of resveratrol.

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Resveratrol facilitates the TyrRS/PARP-1 interaction in an active-site-dependent mannera, Both heat shock (42°C for 30 min) and tunicamycin-treatment (10 μg/ml, ER stress) facilitated the nuclear translocation of TyrRS and activation of PARP-1. b, Resveratrol or serum starvation facilitate TyrRS interaction with PARP-1 and Tyr-SA prevents this interaction. ZZ-PARP-1 was immunoprecipiated with IgG from HeLa cells treated with RSV or serum starvation alone or in combination with Tyr-SA. c, Resveratrol or serum starvation mediated PARP-1 activation is blocked only by Tyr-SA and not by Gly-SA. d, TyrRS interacts directly with PARP-1. HeLa cell lysate after RSV treatment (5 μM, 30 min) was divided into three parts and treated with PARG and catalytically inactive PARG-MT. PARP-1 was immunoprecipitated and analyzed for TyrRS interaction. e. Model illustrating the mechanism of RSV mediated TyrRS interaction with PARP-1 and subsequent release after auto-PARylation. f. Ni-NTA pull-down of N- and C-terminal fragments of PARP-1 overexpressed in E. coli demonstrated that TyrRS interacts with the C-terminal region of PARP-1. g, Only the full-length TyrRS (1-528) and none of the various fragments of TyrRS (mini-TyrRS (1-364), ΔN-TyrRS (228-528) or the C-domain (328-528)) interacts with PARP-1. h, Coomassie blue staining of a gel showing the total protein input in for the experiment of Extended Data Figure 3g.
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Figure 7: Resveratrol facilitates the TyrRS/PARP-1 interaction in an active-site-dependent mannera, Both heat shock (42°C for 30 min) and tunicamycin-treatment (10 μg/ml, ER stress) facilitated the nuclear translocation of TyrRS and activation of PARP-1. b, Resveratrol or serum starvation facilitate TyrRS interaction with PARP-1 and Tyr-SA prevents this interaction. ZZ-PARP-1 was immunoprecipiated with IgG from HeLa cells treated with RSV or serum starvation alone or in combination with Tyr-SA. c, Resveratrol or serum starvation mediated PARP-1 activation is blocked only by Tyr-SA and not by Gly-SA. d, TyrRS interacts directly with PARP-1. HeLa cell lysate after RSV treatment (5 μM, 30 min) was divided into three parts and treated with PARG and catalytically inactive PARG-MT. PARP-1 was immunoprecipitated and analyzed for TyrRS interaction. e. Model illustrating the mechanism of RSV mediated TyrRS interaction with PARP-1 and subsequent release after auto-PARylation. f. Ni-NTA pull-down of N- and C-terminal fragments of PARP-1 overexpressed in E. coli demonstrated that TyrRS interacts with the C-terminal region of PARP-1. g, Only the full-length TyrRS (1-528) and none of the various fragments of TyrRS (mini-TyrRS (1-364), ΔN-TyrRS (228-528) or the C-domain (328-528)) interacts with PARP-1. h, Coomassie blue staining of a gel showing the total protein input in for the experiment of Extended Data Figure 3g.

Mentions: Associated with a previous study12, a distinct TyrRS-PARP-1 interaction was observed. PARP-1 is a major modulator of NAD+ metabolism and its related signaling13. Because RSV acts through NAD+-dependent proteins14, the TyrRS-PARP-1 interaction was further studied. Given that RSV treatment elicits a stress response2, serum starvation (SS) was used to mimic a general ‘stand-alone’ stress condition so that common signaling pathways, if any, between RSV treatment and a general stress condition, could be compared ex-vivo. Either serum starvation or RSV treatment promoted nuclear translocation of endogenous TyrRS in HeLa cells (Fig. 1c). Translocation was observed under different stress conditions (heat shock and ER stress, Extended Data Fig. 3a), suggesting that TyrRS is a general stress transducer. Nuclear translocation of endogenous TyrRS was concomitant with strong auto-PARylation of PARP-1 (PARP-1PAR) (Fig. 1c and Extended Data Fig. 3a).


A human tRNA synthetase is a potent PARP1-activating effector target for resveratrol.

Sajish M, Schimmel P - Nature (2014)

Resveratrol facilitates the TyrRS/PARP-1 interaction in an active-site-dependent mannera, Both heat shock (42°C for 30 min) and tunicamycin-treatment (10 μg/ml, ER stress) facilitated the nuclear translocation of TyrRS and activation of PARP-1. b, Resveratrol or serum starvation facilitate TyrRS interaction with PARP-1 and Tyr-SA prevents this interaction. ZZ-PARP-1 was immunoprecipiated with IgG from HeLa cells treated with RSV or serum starvation alone or in combination with Tyr-SA. c, Resveratrol or serum starvation mediated PARP-1 activation is blocked only by Tyr-SA and not by Gly-SA. d, TyrRS interacts directly with PARP-1. HeLa cell lysate after RSV treatment (5 μM, 30 min) was divided into three parts and treated with PARG and catalytically inactive PARG-MT. PARP-1 was immunoprecipitated and analyzed for TyrRS interaction. e. Model illustrating the mechanism of RSV mediated TyrRS interaction with PARP-1 and subsequent release after auto-PARylation. f. Ni-NTA pull-down of N- and C-terminal fragments of PARP-1 overexpressed in E. coli demonstrated that TyrRS interacts with the C-terminal region of PARP-1. g, Only the full-length TyrRS (1-528) and none of the various fragments of TyrRS (mini-TyrRS (1-364), ΔN-TyrRS (228-528) or the C-domain (328-528)) interacts with PARP-1. h, Coomassie blue staining of a gel showing the total protein input in for the experiment of Extended Data Figure 3g.
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Figure 7: Resveratrol facilitates the TyrRS/PARP-1 interaction in an active-site-dependent mannera, Both heat shock (42°C for 30 min) and tunicamycin-treatment (10 μg/ml, ER stress) facilitated the nuclear translocation of TyrRS and activation of PARP-1. b, Resveratrol or serum starvation facilitate TyrRS interaction with PARP-1 and Tyr-SA prevents this interaction. ZZ-PARP-1 was immunoprecipiated with IgG from HeLa cells treated with RSV or serum starvation alone or in combination with Tyr-SA. c, Resveratrol or serum starvation mediated PARP-1 activation is blocked only by Tyr-SA and not by Gly-SA. d, TyrRS interacts directly with PARP-1. HeLa cell lysate after RSV treatment (5 μM, 30 min) was divided into three parts and treated with PARG and catalytically inactive PARG-MT. PARP-1 was immunoprecipitated and analyzed for TyrRS interaction. e. Model illustrating the mechanism of RSV mediated TyrRS interaction with PARP-1 and subsequent release after auto-PARylation. f. Ni-NTA pull-down of N- and C-terminal fragments of PARP-1 overexpressed in E. coli demonstrated that TyrRS interacts with the C-terminal region of PARP-1. g, Only the full-length TyrRS (1-528) and none of the various fragments of TyrRS (mini-TyrRS (1-364), ΔN-TyrRS (228-528) or the C-domain (328-528)) interacts with PARP-1. h, Coomassie blue staining of a gel showing the total protein input in for the experiment of Extended Data Figure 3g.
Mentions: Associated with a previous study12, a distinct TyrRS-PARP-1 interaction was observed. PARP-1 is a major modulator of NAD+ metabolism and its related signaling13. Because RSV acts through NAD+-dependent proteins14, the TyrRS-PARP-1 interaction was further studied. Given that RSV treatment elicits a stress response2, serum starvation (SS) was used to mimic a general ‘stand-alone’ stress condition so that common signaling pathways, if any, between RSV treatment and a general stress condition, could be compared ex-vivo. Either serum starvation or RSV treatment promoted nuclear translocation of endogenous TyrRS in HeLa cells (Fig. 1c). Translocation was observed under different stress conditions (heat shock and ER stress, Extended Data Fig. 3a), suggesting that TyrRS is a general stress transducer. Nuclear translocation of endogenous TyrRS was concomitant with strong auto-PARylation of PARP-1 (PARP-1PAR) (Fig. 1c and Extended Data Fig. 3a).

Bottom Line: Resveratrol is reported to extend lifespan and provide cardio-neuro-protective, anti-diabetic, and anti-cancer effects by initiating a stress response that induces survival genes.Because human tyrosyl transfer-RNA (tRNA) synthetase (TyrRS) translocates to the nucleus under stress conditions, we considered the possibility that the tyrosine-like phenolic ring of resveratrol might fit into the active site pocket to effect a nuclear role.In contrast to functionally diverse tRNA synthetase catalytic s created by alternative splicing events that ablate active sites, here a non-spliced TyrRS catalytic reveals a new PARP1- and NAD(+)-dependent dimension to the physiological mechanism of resveratrol.

View Article: PubMed Central - PubMed

Affiliation: The Skaggs Institute for Chemical Biology, The Scripps Laboratories for tRNA Synthetase Research, Department of Molecular and Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.

ABSTRACT
Resveratrol is reported to extend lifespan and provide cardio-neuro-protective, anti-diabetic, and anti-cancer effects by initiating a stress response that induces survival genes. Because human tyrosyl transfer-RNA (tRNA) synthetase (TyrRS) translocates to the nucleus under stress conditions, we considered the possibility that the tyrosine-like phenolic ring of resveratrol might fit into the active site pocket to effect a nuclear role. Here we present a 2.1 Å co-crystal structure of resveratrol bound to the active site of TyrRS. Resveratrol ifies the catalytic activity and redirects TyrRS to a nuclear function, stimulating NAD(+)-dependent auto-poly-ADP-ribosylation of poly(ADP-ribose) polymerase 1 (PARP1). Downstream activation of key stress signalling pathways are causally connected to TyrRS-PARP1-NAD(+) collaboration. This collaboration is also demonstrated in the mouse, and is specifically blocked in vivo by a resveratrol-displacing tyrosyl adenylate analogue. In contrast to functionally diverse tRNA synthetase catalytic s created by alternative splicing events that ablate active sites, here a non-spliced TyrRS catalytic reveals a new PARP1- and NAD(+)-dependent dimension to the physiological mechanism of resveratrol.

Show MeSH
Related in: MedlinePlus