Nitric oxide regulates the 26S proteasome in vascular smooth muscle cells

Kapadia, Muneera R.; Eng, Jason W.-L.; Jiang, Qun; Stoyanovsky, Detcho A.; Kibbe, Melina R.

doi:10.1016/j.niox.2009.02.005

Cited by 58 publications

(53 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…2E and supplemental Table S5). These results suggest extensive regulation of these processes by reversible S-nitrosylation and they may also help explain some of the known biological effects of NO, such as inhibition of protein translation (43) and modulation of proteasome function (44). We note that many of the proteins and pathways that we identified in the analysis of the macrophage nitrosoproteome overlap with the recently identified iNOS interactome (45) consistent with recent evidence that nitrosylation occurs in a localized manner and facilitated by protein-protein interactions (3, 46 -49).…”

Section: Discussionmentioning

confidence: 76%

A Substrate Trapping Approach Identifies Proteins Regulated by Reversible S-nitrosylation

Ben-Lulu

Ziv

Admon

et al. 2014

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

Protein S-nitrosylation, the nitric oxide-mediated posttranslational modification of cysteine residues, has emerged as an important regulatory mechanism in diverse cellular processes. Yet, knowledge about the Snitrosoproteome in different cell types and cellular contexts is still limited and many questions remain regarding the precise roles of protein S-nitrosylation and denitrosylation. Here we present a novel strategy to identify reversibly nitrosylated proteins. Our approach is based on nitrosothiol capture and enrichment using a thioredoxin trap mutant, followed by protein identification by mass spectrometry. Employing this approach, we identified more than 400 putative nitroso-proteins in S-nitrosocysteinetreated human monocytes and about 200 nitrosylation substrates in endotoxin and cytokine-stimulated mouse macrophages. The large majority of these represent novel nitrosylation targets and they include many proteins with key functions in cellular homeostasis and signaling. Biochemical and functional experiments in vitro and in cells validated the proteomic results and further suggested a role for thioredoxin in the denitrosylation and activation of inducible nitric oxide synthase and the protein kinase MEK1. Our findings contribute to a better understanding of the macrophage S-nitrosoproteome and the role of thioredoxin-mediated denitrosylation in nitric oxide signaling. The approach described here may prove generally useful for the identification and exploration of nitrosoproteomes under various physiological and pathophysiological conditions. Molecular & Cellular

show abstract

Section: Discussionmentioning

confidence: 76%

A Substrate Trapping Approach Identifies Proteins Regulated by Reversible S-nitrosylation

Ben-Lulu

Ziv

Admon

et al. 2014

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

show abstract

“…NO has been shown to suppress proteasome activity in VSMC by S-nitrosylation, an effect generated through a direct contact of abundant NO with the proteasomes [34]. We asked whether direct interaction of NO with 26S proteasomes would lead to 26S proteasome suppression in endothelial cells.…”

Section: Resultsmentioning

confidence: 99%

“…NO has been found to inhibit the 26S proteasome, resulting in diminished p53 degradation [32] or loss of cell viability [33]. The suppressive effect is mediated by S-nitrosylation and reduced proteasomal subunit expression in vascular smooth muscle cells (VSMC) [34]. However, others report that NO enhances proteasome activity [35] and that the activation promoted by NO donor (DETA-NONOate) is thought to reduce neutral ceramidase [36], [37] or to protect the endothelial cell from damage induced by H 2 O 2 [38].…”

Section: Introductionmentioning

confidence: 99%

Identification of Nitric Oxide as an Endogenous Inhibitor of 26S Proteasomes in Vascular Endothelial Cells

Liu

Zhang

et al. 2014

PLoS ONE

View full text Add to dashboard Cite

The 26S proteasome plays a fundamental role in almost all eukaryotic cells, including vascular endothelial cells. However, it remains largely unknown how proteasome functionality is regulated in the vasculature. Endothelial nitric oxide (NO) synthase (eNOS)-derived NO is known to be essential to maintain endothelial homeostasis. The aim of the present study was to establish the connection between endothelial NO and 26S proteasome functionality in vascular endothelial cells. The 26S proteasome reporter protein levels, 26S proteasome activity, and the O-GlcNAcylation of Rpt2, a key subunit of the proteasome regulatory complex, were assayed in 26S proteasome reporter cells, human umbilical vein endothelial cells (HUVEC), and mouse aortic tissues isolated from 26S proteasome reporter and eNOS knockout mice. Like the other selective NO donors, NO derived from activated eNOS (by pharmacological and genetic approach) increased O-GlcNAc modification of Rpt2, reduced proteasome chymotrypsin-like activity, and caused 26S proteasome reporter protein accumulation. Conversely, inactivation of eNOS reversed all the effects. SiRNA knockdown of O-GlcNAc transferase (OGT), the key enzyme that catalyzes protein O-GlcNAcylation, abolished NO-induced effects. Consistently, adenoviral overexpression of O-GlcNAcase (OGA), the enzyme catalyzing the removal of the O-GlcNAc group, mimicked the effects of OGT knockdown. Finally, compared to eNOS wild type aortic tissues, 26S proteasome reporter mice lacking eNOS exhibited elevated 26S proteasome functionality in parallel with decreased Rpt2 O-GlcNAcylation, without changing the levels of Rpt2 protein. In conclusion, the eNOS-derived NO functions as a physiological suppressor of the 26S proteasome in vascular endothelial cells.

show abstract

“…The effects of temporary inhibition of the proteasome in the vasculature are also well-known, and are similar to those of NO administration following balloon injury. They include prevention of neointimal hyperplasia, increased cellular apoptosis, reduction in macrophage infiltration, and accumulation of ubiquitinated proteins [22,23]. The buildup of ubiquitinated proteins due to chronic inhibition of the proteasome has been linked to a number of cardiovascular dysfunctions, including cardiomyopathy, unstable atherosclerotic plaques, and increased intimal hyperplasia of vein grafts [24][25][26][27][28][29].…”

Section: Discussionmentioning

confidence: 99%

“…All animal procedures were performed in accordance with the Guide for the Care and Use of Laboratory Animals published by the National Institutes of Health (NIH Publication 85- 23,1996) and approved by the Northwestern University Animal Care and Use Committee. Male Sprague-Dawley rats weighing between 350 and 400 g were anesthetized with inhaled isoflurane (0.5-3%).…”

Section: Animal Surgerymentioning

confidence: 99%

Nitric Oxide Inhibits Vascular Smooth Muscle Cell Proliferation and Neointimal Hyperplasia by Increasing the Ubiquitination and Degradation of UbcH10

Tsihlis

Oustwani

Vavra

et al. 2011

Cell Biochem Biophys

Self Cite

View full text Add to dashboard Cite

Nitric oxide (NO) limits formation of neointimal hyperplasia in animal models of arterial injury in large part by inhibiting vascular smooth muscle cell (VSMC) proliferation through cell cycle arrest. The ubiquitin-conjugating enzyme UbcH10 is responsible for ubiquitinating cell cycle proteins for proper exit from mitosis. We hypothesize that NO prevents VSMC proliferation, and hence neointimal hyperplasia, by decreasing levels of UbcH10. Western blotting and immunofluorescent staining showed that NO reduced UbcH10 levels in a concentration-dependent manner in VSMC harvested from the abdominal aortas of Sprague-Dawley rats. Treatment with NO or siRNA to UbcH10 decreased both UbcH10 levels and VSMC proliferation (P<0.001), while increasing UbcH10 levels by plasmid transfection or angiotensin II stimulation increased VSMC proliferation to 150% (P=0.008) and 212% (P=0.002) of control, respectively. Immunofluorescent staining of balloon-injured rat carotid arteries showed a ~4-fold increase in UbcH10 levels, which was profoundly decreased following treatment with NO. Western blotting of carotid artery lysates showed no UbcH10 in uninjured vessels, a substantial increase in the injury alone group, and a significant decrease in the injury+NO group (~3-fold reduction versus injury alone). Importantly, in vitro and in vivo, a marked increase in polyubiquitinated UbcH10 was observed in the NO-treated VSMC and carotid arteries, respectively, indicating that NO may be decreasing unmodified UbcH10 levels by increasing its ubiquitination. Central to our hypothesis, we report that NO decreases UbcH10 levels in VSMC in vitro and following arterial injury in vivo in association with increasing polyubiquitinated-UbcH10 levels. These changes in UbcH10 levels correlate with VSMC proliferation and neointimal hyperplasia, making UbcH10 a promising therapeutic target for inhibiting this proliferative disease.

show abstract

Nitric oxide regulates the 26S proteasome in vascular smooth muscle cells

Cited by 58 publications

References 36 publications

A Substrate Trapping Approach Identifies Proteins Regulated by Reversible S-nitrosylation

A Substrate Trapping Approach Identifies Proteins Regulated by Reversible S-nitrosylation

Identification of Nitric Oxide as an Endogenous Inhibitor of 26S Proteasomes in Vascular Endothelial Cells

Nitric Oxide Inhibits Vascular Smooth Muscle Cell Proliferation and Neointimal Hyperplasia by Increasing the Ubiquitination and Degradation of UbcH10

Contact Info

Product

Resources

About