PKG1-modified TSC2 regulates mTORC1 activity to counter adverse cardiac stress

Mark Ranek, Kristen M. Kokkonen-Simon, Anna Chen, Brittany L. Dunkerly-Eyring, Miguel Pinilla Vera, Christian U. Oeing, Chirag H. Patel, Taishi Nakamura, Guangshuo Zhu, Djahida Bedja, Masayuki Sasaki, Ronald J. Holewinski, Jennifer E. Van Eyk, Jonathan Powell, Dong Lee, David A Kass

Research output: Contribution to journalLetter

Abstract

The mechanistic target of rapamycin complex-1 (mTORC1) coordinates regulation of growth, metabolism, protein synthesis and autophagy 1 . Its hyperactivation contributes to disease in numerous organs, including the heart 1,2 , although broad inhibition of mTORC1 risks interference with its homeostatic roles. Tuberin (TSC2) is a GTPase-activating protein and prominent intrinsic regulator of mTORC1 that acts through modulation of RHEB (Ras homologue enriched in brain). TSC2 constitutively inhibits mTORC1; however, this activity is modified by phosphorylation from multiple signalling kinases that in turn inhibits (AMPK and GSK-3β) or stimulates (AKT, ERK and RSK-1) mTORC1 activity 3–9 . Each kinase requires engagement of multiple serines, impeding analysis of their role in vivo. Here we show that phosphorylation or gain- or loss-of-function mutations at either of two adjacent serine residues in TSC2 (S1365 and S1366 in mice; S1364 and S1365 in humans) can bidirectionally control mTORC1 activity stimulated by growth factors or haemodynamic stress, and consequently modulate cell growth and autophagy. However, basal mTORC1 activity remains unchanged. In the heart, or in isolated cardiomyocytes or fibroblasts, protein kinase G1 (PKG1) phosphorylates these TSC2 sites. PKG1 is a primary effector of nitric oxide and natriuretic peptide signalling, and protects against heart disease 10–13 . Suppression of hypertrophy and stimulation of autophagy in cardiomyocytes by PKG1 requires TSC2 phosphorylation. Homozygous knock-in mice that express a phosphorylation-silencing mutation in TSC2 (TSC2(S1365A)) develop worse heart disease and have higher mortality after sustained pressure overload of the heart, owing to mTORC1 hyperactivity that cannot be rescued by PKG1 stimulation. However, cardiac disease is reduced and survival of heterozygote Tsc2 S1365A knock-in mice subjected to the same stress is improved by PKG1 activation or expression of a phosphorylation-mimicking mutation (TSC2(S1365E)). Resting mTORC1 activity is not altered in either knock-in model. Therefore, TSC2 phosphorylation is both required and sufficient for PKG1-mediated cardiac protection against pressure overload. The serine residues identified here provide a genetic tool for bidirectional regulation of the amplitude of stress-stimulated mTORC1 activity.

Original languageEnglish (US)
Pages (from-to)264-269
Number of pages6
JournalNature
Volume566
Issue number7743
DOIs
StatePublished - Feb 14 2019

Fingerprint

Protein Kinases
Phosphorylation
Autophagy
Serine
Heart Diseases
Cardiac Myocytes
Mutation
Phosphotransferases
mechanistic target of rapamycin complex 1
Glycogen Synthase Kinase 3
GTPase-Activating Proteins
Pressure
Natriuretic Peptides
AMP-Activated Protein Kinases
Heterozygote
Growth
Hypertrophy
Intercellular Signaling Peptides and Proteins
Nitric Oxide
Fibroblasts

ASJC Scopus subject areas

  • General

Cite this

Ranek, M., Kokkonen-Simon, K. M., Chen, A., Dunkerly-Eyring, B. L., Vera, M. P., Oeing, C. U., ... Kass, D. A. (2019). PKG1-modified TSC2 regulates mTORC1 activity to counter adverse cardiac stress. Nature, 566(7743), 264-269. https://doi.org/10.1038/s41586-019-0895-y

PKG1-modified TSC2 regulates mTORC1 activity to counter adverse cardiac stress. / Ranek, Mark; Kokkonen-Simon, Kristen M.; Chen, Anna; Dunkerly-Eyring, Brittany L.; Vera, Miguel Pinilla; Oeing, Christian U.; Patel, Chirag H.; Nakamura, Taishi; Zhu, Guangshuo; Bedja, Djahida; Sasaki, Masayuki; Holewinski, Ronald J.; Van Eyk, Jennifer E.; Powell, Jonathan; Lee, Dong; Kass, David A.

In: Nature, Vol. 566, No. 7743, 14.02.2019, p. 264-269.

Research output: Contribution to journalLetter

Ranek, M, Kokkonen-Simon, KM, Chen, A, Dunkerly-Eyring, BL, Vera, MP, Oeing, CU, Patel, CH, Nakamura, T, Zhu, G, Bedja, D, Sasaki, M, Holewinski, RJ, Van Eyk, JE, Powell, J, Lee, D & Kass, DA 2019, 'PKG1-modified TSC2 regulates mTORC1 activity to counter adverse cardiac stress', Nature, vol. 566, no. 7743, pp. 264-269. https://doi.org/10.1038/s41586-019-0895-y
Ranek M, Kokkonen-Simon KM, Chen A, Dunkerly-Eyring BL, Vera MP, Oeing CU et al. PKG1-modified TSC2 regulates mTORC1 activity to counter adverse cardiac stress. Nature. 2019 Feb 14;566(7743):264-269. https://doi.org/10.1038/s41586-019-0895-y
Ranek, Mark ; Kokkonen-Simon, Kristen M. ; Chen, Anna ; Dunkerly-Eyring, Brittany L. ; Vera, Miguel Pinilla ; Oeing, Christian U. ; Patel, Chirag H. ; Nakamura, Taishi ; Zhu, Guangshuo ; Bedja, Djahida ; Sasaki, Masayuki ; Holewinski, Ronald J. ; Van Eyk, Jennifer E. ; Powell, Jonathan ; Lee, Dong ; Kass, David A. / PKG1-modified TSC2 regulates mTORC1 activity to counter adverse cardiac stress. In: Nature. 2019 ; Vol. 566, No. 7743. pp. 264-269.
@article{1e09a2bc2bd44b0bba080fe102ae8caf,
title = "PKG1-modified TSC2 regulates mTORC1 activity to counter adverse cardiac stress",
abstract = "The mechanistic target of rapamycin complex-1 (mTORC1) coordinates regulation of growth, metabolism, protein synthesis and autophagy 1 . Its hyperactivation contributes to disease in numerous organs, including the heart 1,2 , although broad inhibition of mTORC1 risks interference with its homeostatic roles. Tuberin (TSC2) is a GTPase-activating protein and prominent intrinsic regulator of mTORC1 that acts through modulation of RHEB (Ras homologue enriched in brain). TSC2 constitutively inhibits mTORC1; however, this activity is modified by phosphorylation from multiple signalling kinases that in turn inhibits (AMPK and GSK-3β) or stimulates (AKT, ERK and RSK-1) mTORC1 activity 3–9 . Each kinase requires engagement of multiple serines, impeding analysis of their role in vivo. Here we show that phosphorylation or gain- or loss-of-function mutations at either of two adjacent serine residues in TSC2 (S1365 and S1366 in mice; S1364 and S1365 in humans) can bidirectionally control mTORC1 activity stimulated by growth factors or haemodynamic stress, and consequently modulate cell growth and autophagy. However, basal mTORC1 activity remains unchanged. In the heart, or in isolated cardiomyocytes or fibroblasts, protein kinase G1 (PKG1) phosphorylates these TSC2 sites. PKG1 is a primary effector of nitric oxide and natriuretic peptide signalling, and protects against heart disease 10–13 . Suppression of hypertrophy and stimulation of autophagy in cardiomyocytes by PKG1 requires TSC2 phosphorylation. Homozygous knock-in mice that express a phosphorylation-silencing mutation in TSC2 (TSC2(S1365A)) develop worse heart disease and have higher mortality after sustained pressure overload of the heart, owing to mTORC1 hyperactivity that cannot be rescued by PKG1 stimulation. However, cardiac disease is reduced and survival of heterozygote Tsc2 S1365A knock-in mice subjected to the same stress is improved by PKG1 activation or expression of a phosphorylation-mimicking mutation (TSC2(S1365E)). Resting mTORC1 activity is not altered in either knock-in model. Therefore, TSC2 phosphorylation is both required and sufficient for PKG1-mediated cardiac protection against pressure overload. The serine residues identified here provide a genetic tool for bidirectional regulation of the amplitude of stress-stimulated mTORC1 activity.",
author = "Mark Ranek and Kokkonen-Simon, {Kristen M.} and Anna Chen and Dunkerly-Eyring, {Brittany L.} and Vera, {Miguel Pinilla} and Oeing, {Christian U.} and Patel, {Chirag H.} and Taishi Nakamura and Guangshuo Zhu and Djahida Bedja and Masayuki Sasaki and Holewinski, {Ronald J.} and {Van Eyk}, {Jennifer E.} and Jonathan Powell and Dong Lee and Kass, {David A}",
year = "2019",
month = "2",
day = "14",
doi = "10.1038/s41586-019-0895-y",
language = "English (US)",
volume = "566",
pages = "264--269",
journal = "Nature",
issn = "0028-0836",
publisher = "Nature Publishing Group",
number = "7743",

}

TY - JOUR

T1 - PKG1-modified TSC2 regulates mTORC1 activity to counter adverse cardiac stress

AU - Ranek, Mark

AU - Kokkonen-Simon, Kristen M.

AU - Chen, Anna

AU - Dunkerly-Eyring, Brittany L.

AU - Vera, Miguel Pinilla

AU - Oeing, Christian U.

AU - Patel, Chirag H.

AU - Nakamura, Taishi

AU - Zhu, Guangshuo

AU - Bedja, Djahida

AU - Sasaki, Masayuki

AU - Holewinski, Ronald J.

AU - Van Eyk, Jennifer E.

AU - Powell, Jonathan

AU - Lee, Dong

AU - Kass, David A

PY - 2019/2/14

Y1 - 2019/2/14

N2 - The mechanistic target of rapamycin complex-1 (mTORC1) coordinates regulation of growth, metabolism, protein synthesis and autophagy 1 . Its hyperactivation contributes to disease in numerous organs, including the heart 1,2 , although broad inhibition of mTORC1 risks interference with its homeostatic roles. Tuberin (TSC2) is a GTPase-activating protein and prominent intrinsic regulator of mTORC1 that acts through modulation of RHEB (Ras homologue enriched in brain). TSC2 constitutively inhibits mTORC1; however, this activity is modified by phosphorylation from multiple signalling kinases that in turn inhibits (AMPK and GSK-3β) or stimulates (AKT, ERK and RSK-1) mTORC1 activity 3–9 . Each kinase requires engagement of multiple serines, impeding analysis of their role in vivo. Here we show that phosphorylation or gain- or loss-of-function mutations at either of two adjacent serine residues in TSC2 (S1365 and S1366 in mice; S1364 and S1365 in humans) can bidirectionally control mTORC1 activity stimulated by growth factors or haemodynamic stress, and consequently modulate cell growth and autophagy. However, basal mTORC1 activity remains unchanged. In the heart, or in isolated cardiomyocytes or fibroblasts, protein kinase G1 (PKG1) phosphorylates these TSC2 sites. PKG1 is a primary effector of nitric oxide and natriuretic peptide signalling, and protects against heart disease 10–13 . Suppression of hypertrophy and stimulation of autophagy in cardiomyocytes by PKG1 requires TSC2 phosphorylation. Homozygous knock-in mice that express a phosphorylation-silencing mutation in TSC2 (TSC2(S1365A)) develop worse heart disease and have higher mortality after sustained pressure overload of the heart, owing to mTORC1 hyperactivity that cannot be rescued by PKG1 stimulation. However, cardiac disease is reduced and survival of heterozygote Tsc2 S1365A knock-in mice subjected to the same stress is improved by PKG1 activation or expression of a phosphorylation-mimicking mutation (TSC2(S1365E)). Resting mTORC1 activity is not altered in either knock-in model. Therefore, TSC2 phosphorylation is both required and sufficient for PKG1-mediated cardiac protection against pressure overload. The serine residues identified here provide a genetic tool for bidirectional regulation of the amplitude of stress-stimulated mTORC1 activity.

AB - The mechanistic target of rapamycin complex-1 (mTORC1) coordinates regulation of growth, metabolism, protein synthesis and autophagy 1 . Its hyperactivation contributes to disease in numerous organs, including the heart 1,2 , although broad inhibition of mTORC1 risks interference with its homeostatic roles. Tuberin (TSC2) is a GTPase-activating protein and prominent intrinsic regulator of mTORC1 that acts through modulation of RHEB (Ras homologue enriched in brain). TSC2 constitutively inhibits mTORC1; however, this activity is modified by phosphorylation from multiple signalling kinases that in turn inhibits (AMPK and GSK-3β) or stimulates (AKT, ERK and RSK-1) mTORC1 activity 3–9 . Each kinase requires engagement of multiple serines, impeding analysis of their role in vivo. Here we show that phosphorylation or gain- or loss-of-function mutations at either of two adjacent serine residues in TSC2 (S1365 and S1366 in mice; S1364 and S1365 in humans) can bidirectionally control mTORC1 activity stimulated by growth factors or haemodynamic stress, and consequently modulate cell growth and autophagy. However, basal mTORC1 activity remains unchanged. In the heart, or in isolated cardiomyocytes or fibroblasts, protein kinase G1 (PKG1) phosphorylates these TSC2 sites. PKG1 is a primary effector of nitric oxide and natriuretic peptide signalling, and protects against heart disease 10–13 . Suppression of hypertrophy and stimulation of autophagy in cardiomyocytes by PKG1 requires TSC2 phosphorylation. Homozygous knock-in mice that express a phosphorylation-silencing mutation in TSC2 (TSC2(S1365A)) develop worse heart disease and have higher mortality after sustained pressure overload of the heart, owing to mTORC1 hyperactivity that cannot be rescued by PKG1 stimulation. However, cardiac disease is reduced and survival of heterozygote Tsc2 S1365A knock-in mice subjected to the same stress is improved by PKG1 activation or expression of a phosphorylation-mimicking mutation (TSC2(S1365E)). Resting mTORC1 activity is not altered in either knock-in model. Therefore, TSC2 phosphorylation is both required and sufficient for PKG1-mediated cardiac protection against pressure overload. The serine residues identified here provide a genetic tool for bidirectional regulation of the amplitude of stress-stimulated mTORC1 activity.

UR - http://www.scopus.com/inward/record.url?scp=85061233152&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85061233152&partnerID=8YFLogxK

U2 - 10.1038/s41586-019-0895-y

DO - 10.1038/s41586-019-0895-y

M3 - Letter

VL - 566

SP - 264

EP - 269

JO - Nature

JF - Nature

SN - 0028-0836

IS - 7743

ER -