Hydrogen Sulfide-Linked Sulfhydration of NF-κB Mediates Its Antiapoptotic Actions

The Solomon H Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Molecular cell (Impact Factor: 14.02). 01/2012; 45(1):13-24. DOI: 10.1016/j.molcel.2011.10.021
Source: PubMed


Nuclear factor κB (NF-κB) is an antiapoptotic transcription factor. We show that the antiapoptotic actions of NF-κB are mediated by hydrogen sulfide (H(2)S) synthesized by cystathionine gamma-lyase (CSE). TNF-α treatment triples H(2)S generation by stimulating binding of SP1 to the CSE promoter. H(2)S generated by CSE stimulates DNA binding and gene activation of NF-κB, processes that are abolished in CSE-deleted mice. As CSE deletion leads to decreased glutathione levels, resultant oxidative stress may contribute to alterations in CSE mutant mice. H(2)S acts by sulfhydrating the p65 subunit of NF-κB at cysteine-38, which promotes its binding to the coactivator ribosomal protein S3 (RPS3). Sulfhydration of p65 predominates early after TNF-α treatment, then declines and is succeeded by a reciprocal enhancement of p65 nitrosylation. In CSE mutant mice, antiapoptotic influences of NF-κB are markedly diminished. Thus, sulfhydration of NF-κB appears to be a physiologic determinant of its antiapoptotic transcriptional activity.

Download full-text


Available from: Risheng Xu
  • Source
    • " S ) - mediated signaling pathways involved in many physiological and pathophysiological processes ( Abe & Kimura , 1996 ; Eberhardt et al . , 2014 ; Elrod et al . , 2007 ; Greiner et al . , 2013 ; Kabil & Banerjee , 2010 ; Krishnan , Fu , Pappin , & Tonks , 2011 ; Li , Rose , & Moore , 2011 ; Mustafa et al . , 2009 , 2011 ; Paul & Snyder , 2012 ; Sen et al . , 2012 ; Szabo , 2007 ; Vandiver et al . , 2013 ; Yang et al . , 2008 , 2013 ; Zhao , Zhang , Lu , & Wang , 2001 ) ."
    [Show abstract] [Hide abstract]
    ABSTRACT: Protein S-sulfhydration (i.e., converting protein cysteines -SH to persulfides -SSH) is a redox-based posttranslational modification. This reaction plays an important role in signaling pathways mediated by hydrogen sulfide or other reactive sulfane sulfur species. Recently, our laboratories developed a "tag-switch" method which can be used to selectively label and detect protein S-sulfhydrated residues. In this chapter, we provide a comprehensive summary of this method, including the design of the method, preparation of the reagents, validation on small-molecule substrates, as well as applications in protein labeling. Experimental protocols for the use of the method are described in details. © 2015 Elsevier Inc. All rights reserved.
    Full-text · Article · Dec 2015 · Methods in enzymology
  • Source
    • "The essential role of RPS3 in directing NF-jB to a subset of genes makes it a potential target for selective NF-jB inhibition in cancer cells. Moreover, RPS3 was recently revealed as a physiologic determinant of NF-jB-mediated transcription of antiapoptotic genes in macrophages, including Birc3 (encoding cellular inhibitor of apoptosis protein-2, cIAP2), Bcl2l1 (encoding B-cell lymphoma-extra large, Bcl-XL), and Xiap (encoding X-linked inhibitor of apoptosis protein, XIAP) [26]. We recently showed that N-terminal fragments of p65, generated by ectopic expression or pathogen protease cleavage, selectively retard RPS3 nuclear translocation and RPS3-conferred NF-jB gene transcription, without affecting p65 [22] [27]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Caspase-3-mediated p65 cleavage is believed to suppress nuclear factor-kappa B (NF-κB)-mediated anti-apoptotic transactivation in cells undergoing apoptosis. However, only a small percentage of p65 is cleaved during apoptosis, not in proportion to the dramatic reduction in NF-κB transactivation. Here we show that the p65(1-97) fragment generated by Caspase-3 cleavage interferes with ribosomal protein S3 (RPS3), an NF-κB "specifier" subunit, and selectively retards the nuclear translocation of RPS3, thus dampening the RPS3/NF-κB-dependent anti-apoptotic gene expression. Our findings reveal a novel cell fate determination mechanism to ensure cells undergo programed cell death through interfering with the RPS3/NF-κB-conferred anti-apoptotic transcription by the fragment from partial p65 cleavage by activated Caspase-3.
    Full-text · Article · Nov 2015 · FEBS letters
  • Source
    • "The number of reports on the biological activity of H 2 S have increased exponentially in the past fifteen years owing to several factors including the availability of facile assays like methylene blue [4] [5] for detection of H 2 S in tissues; seminal papers demonstrating that H 2 S affected important processes like the regulation of ion channels [6] [7] [8]; animal knockouts of CSE can lead to large physiological changes i.e. the elevation of blood pressure [9]; the introduction of methods for detection of proteins modified by Ssulfuration [1] and the direct or indirect demonstration that the sulfuration of proteins like GAPDH [1], protein tyrosine phospha- tase-1B (PTB1B) [10], NF-κB [11], Cu/Zn superoxide dismutase [12] greatly affect their structure and function. Despite the many reports that H 2 S is involved in almost all physiological and pathophysiological processes, the field is fraught with controversy mainly resulting from difficulties in the accurate measurement of H 2 S in biological tissues. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Hydrogen sulfide (H2S) is produced enzymatically by cystathionine β-synthase (CBS) and cystathionine γ-lyase (CSE), as well as other enzymes in mammalian tissues. These discoveries have led to the crowning of H2S as yet another toxic gas that serves as a gasotransmitter like NO and CO. H2S is thought to exert its biological effects through its reaction with cysteine thiols in proteins, yielding sulfurated thiol (-SSH) derivatives. One of the first proteins shown to be modified by H2S was glyceraldehyde 3-phosphate dehydrogenase (GAPDH) [1] where the S-sulfuration of the active site cysteine (Cys 152) resulted in ~7-fold increase in the activity of the enzyme. In the present study we have attempted to reproduce this result with no success. GAPDH in its reduced, or hydrogen peroxide, or glutathione disulfide, or nitrosonium oxidized forms was reacted with sulfide or polysulfides. Sulfide had no effect on reduced GAPDH activity, while polysulfides inhibited GAPDH to ~42% of control. S-sulfuration of GAPDH occurred at Cys 247 after sulfide treatment, Cys 156 and Cys 247 after polysulfide treatment. No evidence of S-sulfuration at active site Cys 152 was discovered. Both sulfide and polysulfide was able to restore the activity of glutathione disulfide oxidized GAPDH, but not to control untreated levels. Treatment of glutathione disulfide oxidized GAPDH with polysulfide also produced S-sulfuration of Cys 156. Treatment of a C156S mutant of GAPDH with sulfide and polysulfide resulted in S-sulfuration of Cys 152, which also caused a decrease and not an increase in enzymatic activity. Computational chemistry shows S-sulfuration of Cys 156 may affect the position of catalytic Cys 152, raising its pKa by 0.5, which may affect the nucleophilicity of Cys 152. The current study raises significant questions about the reported ability of H2S to activate GAPDH by the sulfuration of its active site thiol, and indicates that polysulfide is a stronger protein S-sulfurating agent than sulfide.
    Full-text · Article · Oct 2015 · Free Radical Biology and Medicine
Show more