S-nitrosylated GAPDH initiates apoptotic cell death by nuclear translocation following Siah1 binding

Abstract

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) influences cytotoxicity, translocating to the nucleus during apoptosis. Here we report a signalling pathway in which nitric oxide (NO) generation that follows apoptotic stimulation elicits S-nitrosylation of GAPDH, which triggers binding to Siah1 (an E3 ubiquitin ligase), nuclear translocation and apoptosis. S-nitrosylation of GAPDH augments its binding to Siah1, whose nuclear localization signal mediates translocation of GAPDH. GAPDH stabilizes Siah1, facilitating its degradation of nuclear proteins. Activation of macrophages by endotoxin and of neurons by glutamate elicits GAPDH-Siah1 binding, nuclear translocation and apoptosis, which are prevented by NO deletion. The NO-S-nitrosylation-GAPDH-Siah1 cascade may represent an important molecular mechanism of cytotoxicity.

Glyceraldehyde 3-phosphate dehydrogenase is a SET-binding protein and regulates cyclin B-cdk1 activity

“…These include the transcriptional regulation of histone gene expression (Zheng et al, 2003), the role of GAPDH in nuclear membrane fusion (Nakagawa et al, 2003), in the modulation of telomere structure (Sundararaj et al, 2004), in the recognition of fraudulent nucleotides in DNA arising from cancer chemotherapy (Krynetski et al, 2003) and in triggering apoptosis (Chuang et al, 2005;Hara et al, 2005).…”

Redox-based regulation of signal transduction: Principles, pitfalls, and promises

“…In the latter scenario, glyceraldehyde-3-dehydrogenase (GADPH) is the target of S-nitrosylation of catalytic site cysteine 150, which in turn causes binding of the E3 ligase, Siah1 to GAPDH, and stabilizes Siah1, which is otherwise rapidly turned over. The SNO-GAPDH-Siah1 complex can then translocate to the nucleus, where Siah1 targets nuclear proteins for degradation [184,185].…”

S-Nitrosylation and uncompetitive/fast off-rate (UFO) drug therapy in neurodegenerative disorders of protein misfolding

“…46,47 Our group first identified the physiological relevance of S-nitrosylation by showing that NO and related RNS exert paradoxical effects via redox-based mechanisms -NO is neuroprotective via S-nitrosylation of NMDA receptors (as well as other subsequently discovered targets, including caspases), and yet can also be neurodestructive by formation of peroxynitrite (or, as later discovered, reaction with additional molecules such as parkin, PDI, GAPDH, and MMP-9) (Figure 1). 6,8,9,12,14,16,17,[19][20][21][22]48 Over the past decade, accumulating evidence has suggested that S-nitrosylation can regulate the biological activity of a great variety of proteins, in some ways akin to phosphorylation. 10,[49][50][51][52][53][54] Chemically, NO is often a good 'leaving group,' facilitating further oxidation of critical thiol to disulfide bonds among neighboring (vicinal) cysteine residues or, via reaction with ROS, to sulfenic (ÀSOH), sulfinic (ÀSO 2 H), or sulfonic (ÀSO 3 H) acid derivatization of the protein.…”

“…Recent findings, however, have shed light on molecular events underlying this relationship. Specifically, we and other groups have recently mounted physiological and chemical evidence that S-nitrosylation enhances (1) neuronal survival by inhibiting the activities of (a) NMDA receptors and (b) caspases, or (2) neuronal cell injury by regulating the (a) ubiquitin E3 ligase activity of parkin, 17,18,20 (b) chaperone and isomerase activities of PDI, 19 (c) nuclear translocation of GAPDH, 21 and (d) activity of MMP-9. 22 In particular, S-nitrosylation of PDI and parkin can regulate protein misfolding and neurotoxicity in models of neurodegenerative disorders, and SNO-PDI, or SNO-parkin has been found in human postmortem brain tissue from patients with neurodegenerative conditions.…”

“…(a) S-nitrosylation of NMDA receptors can positively influence neuronal survival; 6,8-11 (b) S-nitrosylation of cysteine protease caspase blocks apoptotic cell death; [12][13][14][15][16] (c) S-nitrosylation of parkin (a ubiquitin E3 ligase) and protein-disulfide isomerase (PDI, an endoplasmic reticulum (ER) chaperone) can be injurious by effecting accumulation of misfolded proteins in neurodegenerative diseases such as PD, AD, and other conditions; [17][18][19][20] (d) S-nitrosylation of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) formed in the cytoplasm transduces an apoptotic signal into the nucleus of neuronal cells; 21 and (e) S-nitrosylation of matrix metalloproteinase (MMP)-2/9 leads to a unique extracellular pathway contributing to excitotoxic neuronal cell damage. 22 NMDA Receptor-Mediated Glutamatergic Signaling Pathways Induce Ca 2 þ Influx…”