Opposing actions of heat shock protein 90 and 70 regulate nicotinamide adenine dinucleotide phosphate oxidase stability and reactive oxygen species production

Feng Chen, Yanfang Yu, Jin Qian, Yusi Wang, Bo Cheng, Christiana Dimitropoulou, Vijaykumar Surendrakant Patel, Ahmed Chadli, Radu Daniel Rudic, David W Stepp, John D. Catravas, David J Fulton

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

OBJECTIVE-: Excessive reactive oxygen species contribute to vascular dysfunction. We have previously shown that heat shock protein (Hsp90) inhibitors potently suppress Nox 1 to 3 and 5, and the goals of this study were to identify how molecular chaperones regulate Nox function. METHODS AND RESULTS-: In vitro, protein expression of Nox 1 to 2, 5 was decreased by Hsp90 inhibitors in multiple cell types (human pulmonary artery endothelial cells, neutrophils, macrophages, and human saphenous vein). In mice treated with Hsp90 inhibitors, Nox1 expression was reduced in lung along with reduced reactive oxygen species from leukocytes. Elevated reactive oxygen species production in obese (db/db) aorta was suppressed by Hsp90 inhibition. Hsp90 inhibitors did not alter Nox5 micro RNA levels, and proteasome inhibition prevented Nox2 and 5 protein degradation and increased ubiquitin incorporation. Inhibition of Hsp90 upregulated the expression of Hsp70 and Hsp70-bound Nox2, 5 and promoted degradation. Silencing Hsp70 prevented Hsp90 inhibitor-mediated degradation of Nox5. The Hsp70-regulated ubiquitin ligase, carboxyl terminus of Hsp70-interacting protein (CHIP), also bound Nox5 and promoted increased Nox5 ubiquitination and degradation. The chaperone binding and ubiquitination domains of CHIP were required, and the silencing of CHIP blunted Hsp90 inhibitor-mediated degradation of Nox2 and 5. CONCLUSION-: We conclude that Hsp90 binds to and regulates Nox protein stability. These actions are opposed by Hsp70 and CHIP, which promote the ubiquitination and degradation of Nox proteins and reduce reactive oxygen species production.

Original languageEnglish (US)
Pages (from-to)2989-2999
Number of pages11
JournalArteriosclerosis, thrombosis, and vascular biology
Volume32
Issue number12
DOIs
StatePublished - Dec 1 2012

Fingerprint

HSP90 Heat-Shock Proteins
HSP70 Heat-Shock Proteins
NADP
Reactive Oxygen Species
Oxidoreductases
Ubiquitination
Ubiquitin
Proteins
Proteolysis
Molecular Chaperones
Protein Stability
Saphenous Vein
Proteasome Endopeptidase Complex
Ligases
Heat-Shock Proteins
MicroRNAs
Pulmonary Artery
Blood Vessels
Aorta
Neutrophils

Keywords

  • CHIP
  • Hsp70
  • Hsp90
  • NADPH oxidase
  • Reactive oxygen species
  • inflammation
  • vascular biology

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

Opposing actions of heat shock protein 90 and 70 regulate nicotinamide adenine dinucleotide phosphate oxidase stability and reactive oxygen species production. / Chen, Feng; Yu, Yanfang; Qian, Jin; Wang, Yusi; Cheng, Bo; Dimitropoulou, Christiana; Patel, Vijaykumar Surendrakant; Chadli, Ahmed; Rudic, Radu Daniel; Stepp, David W; Catravas, John D.; Fulton, David J.

In: Arteriosclerosis, thrombosis, and vascular biology, Vol. 32, No. 12, 01.12.2012, p. 2989-2999.

Research output: Contribution to journalArticle

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abstract = "OBJECTIVE-: Excessive reactive oxygen species contribute to vascular dysfunction. We have previously shown that heat shock protein (Hsp90) inhibitors potently suppress Nox 1 to 3 and 5, and the goals of this study were to identify how molecular chaperones regulate Nox function. METHODS AND RESULTS-: In vitro, protein expression of Nox 1 to 2, 5 was decreased by Hsp90 inhibitors in multiple cell types (human pulmonary artery endothelial cells, neutrophils, macrophages, and human saphenous vein). In mice treated with Hsp90 inhibitors, Nox1 expression was reduced in lung along with reduced reactive oxygen species from leukocytes. Elevated reactive oxygen species production in obese (db/db) aorta was suppressed by Hsp90 inhibition. Hsp90 inhibitors did not alter Nox5 micro RNA levels, and proteasome inhibition prevented Nox2 and 5 protein degradation and increased ubiquitin incorporation. Inhibition of Hsp90 upregulated the expression of Hsp70 and Hsp70-bound Nox2, 5 and promoted degradation. Silencing Hsp70 prevented Hsp90 inhibitor-mediated degradation of Nox5. The Hsp70-regulated ubiquitin ligase, carboxyl terminus of Hsp70-interacting protein (CHIP), also bound Nox5 and promoted increased Nox5 ubiquitination and degradation. The chaperone binding and ubiquitination domains of CHIP were required, and the silencing of CHIP blunted Hsp90 inhibitor-mediated degradation of Nox2 and 5. CONCLUSION-: We conclude that Hsp90 binds to and regulates Nox protein stability. These actions are opposed by Hsp70 and CHIP, which promote the ubiquitination and degradation of Nox proteins and reduce reactive oxygen species production.",
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T1 - Opposing actions of heat shock protein 90 and 70 regulate nicotinamide adenine dinucleotide phosphate oxidase stability and reactive oxygen species production

AU - Chen, Feng

AU - Yu, Yanfang

AU - Qian, Jin

AU - Wang, Yusi

AU - Cheng, Bo

AU - Dimitropoulou, Christiana

AU - Patel, Vijaykumar Surendrakant

AU - Chadli, Ahmed

AU - Rudic, Radu Daniel

AU - Stepp, David W

AU - Catravas, John D.

AU - Fulton, David J

PY - 2012/12/1

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N2 - OBJECTIVE-: Excessive reactive oxygen species contribute to vascular dysfunction. We have previously shown that heat shock protein (Hsp90) inhibitors potently suppress Nox 1 to 3 and 5, and the goals of this study were to identify how molecular chaperones regulate Nox function. METHODS AND RESULTS-: In vitro, protein expression of Nox 1 to 2, 5 was decreased by Hsp90 inhibitors in multiple cell types (human pulmonary artery endothelial cells, neutrophils, macrophages, and human saphenous vein). In mice treated with Hsp90 inhibitors, Nox1 expression was reduced in lung along with reduced reactive oxygen species from leukocytes. Elevated reactive oxygen species production in obese (db/db) aorta was suppressed by Hsp90 inhibition. Hsp90 inhibitors did not alter Nox5 micro RNA levels, and proteasome inhibition prevented Nox2 and 5 protein degradation and increased ubiquitin incorporation. Inhibition of Hsp90 upregulated the expression of Hsp70 and Hsp70-bound Nox2, 5 and promoted degradation. Silencing Hsp70 prevented Hsp90 inhibitor-mediated degradation of Nox5. The Hsp70-regulated ubiquitin ligase, carboxyl terminus of Hsp70-interacting protein (CHIP), also bound Nox5 and promoted increased Nox5 ubiquitination and degradation. The chaperone binding and ubiquitination domains of CHIP were required, and the silencing of CHIP blunted Hsp90 inhibitor-mediated degradation of Nox2 and 5. CONCLUSION-: We conclude that Hsp90 binds to and regulates Nox protein stability. These actions are opposed by Hsp70 and CHIP, which promote the ubiquitination and degradation of Nox proteins and reduce reactive oxygen species production.

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KW - Reactive oxygen species

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KW - vascular biology

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