Cross Talk Between S -Nitrosylation and Phosphorylation Involving Kinases and Nitrosylases

HomeCirculation ResearchVol. 122, No. 11Cross Talk Between S-Nitrosylation and Phosphorylation Involving Kinases and Nitrosylases Free AccessEditorialPDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyReddit Jump toFree AccessEditorialPDF/EPUBCross Talk Between S-Nitrosylation and Phosphorylation Involving Kinases and Nitrosylases Hua-Lin Zhou, Colin T. Stomberski and Jonathan S. Stamler Hua-Lin ZhouHua-Lin Zhou From the Department of Medicine, Institute for Transformative Molecular Medicine, University Hospitals Cleveland Medical Center (H.-L.Z., C.T.S., J.S.S.) Search for more papers by this author , Colin T. StomberskiColin T. Stomberski From the Department of Medicine, Institute for Transformative Molecular Medicine, University Hospitals Cleveland Medical Center (H.-L.Z., C.T.S., J.S.S.) Department of Biochemistry (C.T.S., J.S.S.) Search for more papers by this author and Jonathan S. StamlerJonathan S. Stamler From the Department of Medicine, Institute for Transformative Molecular Medicine, University Hospitals Cleveland Medical Center (H.-L.Z., C.T.S., J.S.S.) Department of Biochemistry (C.T.S., J.S.S.) Case Western Reserve University, OH; Harrington Discovery Institute, University Hospitals Cleveland Medical Center, OH (J.S.S.). Search for more papers by this author Originally published25 May 2018https://doi.org/10.1161/CIRCRESAHA.118.313109Circulation Research. 2018;122:1485–1487GSK3 (glycogen synthase kinase-3)—one of the busiest kinases in cells—phosphorylates >100 known substrates. Typical substrates of GSK3 contain a phosphorylated priming sequence, S/T-X-X-X-S/T(P), where the GSK3-targeted serine/threonine lies 4 residues N-terminal to a phosphorylated serine/threonine. The large number of GSK3 substrates and their location within different cellular compartments raise the question of how selectivity and specificity of GSK3/substrate interactions are achieved. In this issue, Wang et al1 demonstrate that S-nitrosylation of GSK3β inhibits its activity in the cytosol and that S-nitrosylated GSK3β (SNO-GSK3β) translocates to the nucleus to phosphorylate nuclear substrates. Wang et al thus uncover novel interplay between S-nitrosylation and phosphorylation that subserves GSK3β targeting of substrates. Interestingly, this interplay may be relevant in heart failure.Article, see p 1517S-Nitrosylation Directly Suppresses Activity of Protein KinasesIn many cases, cross talk between S-nitrosylation and phosphorylation is manifested by S-nitrosylation–mediated suppression of protein kinase activity (Figure [A]). GRK2 (G protein-coupled receptor kinase 2) is a prime example.2 Specifically, GRK2-mediated phosphorylation of the β-AR (β-adrenergic receptor) at serine and threonine residues in β-AR’s arrestin-binding domain promotes β-arrestin binding, which prevents further activation of G proteins and initiates receptor internalization and desensitization. β-ARs do not, however, desensitize under normal physiological conditions because receptor-mediated kinase activation is followed by S-nitrosylation of a critical Cys (Cys340) within the activation loop of GRK2, which inhibits kinase activity.2 β-AR–coupled, endothelial NOS (NO synthase)–mediated S-nitrosylation thereby prevents receptor inactivation.3 There are many other examples of S-nitrosylation inhibiting phosphorylation-based signal transduction through induced conformational changes in kinases or by blocking their autophosphorylation: PKC (protein kinase C),4 AKT (protein kinase B),5 IKK (IκB kinase),6 and CaMKII (calcium/calmodulin-dependent protein kinase II)7 are to name but a few.Download figureDownload PowerPointFigure. S-nitrosylation and phosphorylation cross talk. A, S-nitrosylation directly inhibits kinase activity. B, S-nitrosylation disrupts kinase–substrate interaction. C, S-nitrosylation changes GSK3β (glycogen synthase kinase-3) localization and determines substrate specificity. D, Phosphorylation (P) and S-nitrosylation alternatively modify β-arr1 (β-arrestin1) and β-arr2 (β-arrestin2) to regulate GPCR internalization. E, S-nitrosylation converts CDK5 (cyclin-dependent kinase 5) from kinase to nitrosylase. AKT indicates protein kinase B; ASK, apoptosis signal-regulating kinase; β-AR, β-adrenergic receptor; CaMKII, calcium/calmodulin-dependent protein kinase II; eNOS, endothelial NO synthase; ERK, extracellular signal-regulated kinase; GRK2, G protein-coupled receptor kinase 2; IKK, IκB kinase; JNK, Jun N-terminal kinase; P, phosphorylation; PKC, protein kinase C; and SNO, S-nitrosothiol (S-nitrosylation).S-Nitrosylation Affects Kinase-Substrate SpecificityProductive interactions between kinases and their substrates identify bona fide phosphorylation sites (among many thousands of candidate sites in the proteome). S-nitrosylation of protein kinases can influence these kinase/substrate interactions. This operating principle is illustrated in the cases of ASK1 (apoptosis signal-regulating kinase-1) and JNK (Jun N-terminal kinases; Figure [B]), which undergo S-nitrosylation in response to apoptotic stimuli.8,9 For example, interferon-γ–induced S-nitrosylation of ASK1 at Cys869 inhibits the binding of ASK1 to its principal downstream effectors MKK3 (mitogen-activated protein kinase kinase 3) and MKK6 (mitogen-activated protein kinase kinase 6), attenuating apoptotic cell death.8S-nitrosylation of Cys116 in JNK inhibits JNK-mediated phosphorylation and transactivation of c-Jun (transcription factor AP-1) by disrupting the interaction between JNK and c-Jun.9 Thus, S-nitrosylation provides a mechanism to regulate substrate recognition in physiological context.Subcellular compartmentalization provides another commonly exploited mechanism to achieve substrate specificity; compartmentalization restricts kinases to a subset of substrates and increases local kinase concentrations. The present study illustrates elegantly how S-nitrosylation determines substrate specificity by changing the localization of GSK3β1 (Figure [C]). GSK3β is traditionally considered to be a cytosolic protein; however, GSK3β is also found in the mitochondria and nucleus, as well as in other subcellular compartments. Although numerous nuclear substrates of GSK3β, including transcriptional factors and epigenetic regulators, have been identified, regulation of GSK3β nuclear localization is poorly understood. Wang et al1 found that S-nitrosylation of GSK3β causes GSK3β translocation to the nucleus where it phosphorylates nuclear substrates. Interestingly, many nuclear substrates of GSK3β contain an S/T-P motif rather than the prototypical priming sequence S/T-X-X-X-S/T(P). This nuance may be because of an S-nitrosylation–induced conformational change in GSK3β that shifts structure-based recognition from the canonical priming site to an alternate phospho-targeting motif. More studies are needed to determine whether S-nitrosylation of GSK3β has evolved to provide a mechanism for recognition of the nuclear S/T-P motif.S-Nitrosylation and Phosphorylation Selectively Modify Different Protein IsoformsProtein isoforms provide a natural mechanism for divergence within signaling networks. Differential modification of protein isoforms by S-nitrosylation versus phosphorylation can fine-tune cellular signaling. Post-translational modification of β-arrestins illustrates this principle (Figure [D]). β-arrestins are versatile, multifunctional scaffold proteins that are best known for their ability to internalize and desensitize GPCRs (G protein-coupled receptors). β-arrestin1 and β-arrestin2 can be both phosphorylated (Ser412 in β-arrestin1 and Thr276, Ser361, and Thr383 in β-arrestin2) and nitrosylated (Cys251 in β-arrestin1 and Cys253 and Cys410 in β-arrestin2) at multiple loci. Stimulus-coupled dephosphorylation of Ser412 in β-arrestin1 is requisite for clathrin-mediated β2-AR internalization. Interestingly, Ser412 in β-arrestin1 is replaced by a highly conserved Cys410 in β-arrestin2, and S-nitrosylation by endothelial NOS of Cys410 in β-arrestin2 promotes its binding to clathrin, thereby accelerating β2-AR internalization.10 Thus, phosphorylation and nitrosylation subserve classic clathrin-mediated internalization of GPCRs by β-arrestin1 and β-arrestin2, respectively. Inasmuch as β-arrestin1 and β-arrestin2 are ubiquitously distributed, these data support the idea of functional cross talk between nitrosylation and phosphorylation in wide-ranging cellular functions.There are 2 isoforms of GSK3 (GSK3α and GSK3β) in mammals. Although GSK3α and GSK3β are ubiquitously expressed and share numerous substrates, they mediate different biological functions in response to cellular stimuli. GSK3β is modified by S-nitrosylation at Cys76, Cys199, and Cys317; GSK3α contains Cys199 and Cys317 but lacks Cys76.1 This is somewhat akin to β-arrestin1 and β-arrestin2, which share 1 SNO site (Cys251/253) but not a second (C410 is present only in β-arrestin2). This raises the question as to whether S-nitrosylation can selectively modify GSK3α and GSK3β to mediate distinct cellular functions.S-Nitrosylation Converts a Protein Kinase Into a Protein NitrosylaseThe machinery for enzymatic S-nitrosylation involves 3 classes of enzymes operating coordinately: NOS, SNO synthases, and transnitrosylases.11 Transnitrosylases mediate thiol-based transfer of NO groups between proteins. Most transnitrosylases have canonical functions apart from their transnitrosylase activity. S-nitrosylation is thus a prerequisite to convert proteins into nascent nitrosylases; this principle is illustrated by GAPDH—a prototypic transnitrosylase.12S-nitrosylation of GAPDH abolishes its catalytic activity and promotes binding to the E3 ubiquitin ligase Siah, which subsequently facilitates translocation of SNO-GAPDH to the nucleus. In the nucleus, SNO-GAPDH acts as a transnitrosylase for numerous nuclear proteins, including the deacetylating enzyme SIRT1 (sirtuin-1), HDAC2 (histone deacetylase-2), and DNA-PK (DNA-activated protein kinase),12 thereby regulating the transcriptional and metabolic activity of the cell.CDK5 (cyclin-dependent kinase 5) is another elegant example of this principle. CDK5 is a proline-directed kinase that phosphorylates serines/threonines immediately upstream of proline residues. CDK5 is implicated in neuronal injury through activation of Drp1 (dynamin-1-like protein), which induces mitochondrial fission. Interestingly, Drp1 can be activated by both phosphorylation and S-nitrosylation. But whereas S-nitrosylation of Drp1-Cys644 mediates mitochondrial fission in Alzheimer (and Huntington) models,13 Ser585 phosphorylation mediates mitochondrial fission in NMDA-induced injury.14 Notably, in Alzheimer disease, CDK5 is S-nitrosylated at an essential Cys83, which resides within the ATP-binding pocket of the kinase, thereby inhibiting kinase activity; SNO-CDK5 is thus converted from a kinase into a Drp1 transnitrosylase, possibly mediating synaptic damage15 (Figure [E]).Wang et al1 show here that S-nitrosylation of GSK3β promotes its trafficking into the nucleus of myocardial cells, and it is well known that multiple nuclear phospho-substrates of GSK3β, including NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells), p53, STAT3 (signal transducer and activator of transcription 3), and Sirt1, can also be modified by S-nitrosylation. It seems likely that SNO-GSK3β may act as a nuclear transnitrosylase to mediate the S-nitrosylation of at least some of these substrates in myocardium.Sources of FundingThis work was supported by National Institutes of Health grants HL075443, HL128192, and HL126900.DisclosuresNone.FootnotesThe opinions expressed in this article are not necessarily those of the editors or of the American Heart Association.Correspondence to Jonathan S. Stamler, MD, Department of Medicine, Institute for Transformative Molecular Medicine, Case Western Reserve University, Wolstein Research Bldg 4129, 2103 Cornell Rd, Cleveland, OH 44106. E-mail [email protected]References1. Wang S-B, Venkatraman V, Crowgey EL, Liu T, Fu Z, Holewinski R, Ranek M, Kass DA, O’Rourke B, Van Eyk JE. Protein S-nitrosylation controls glycogen synthase kinase 3β function independent of its phosphorylation state.Circ Res. 2018; 122:1517–1531. doi: 10.1161/CIRCRESAHA.118.312789LinkGoogle Scholar2. Whalen EJ, Foster MW, Matsumoto A, Ozawa K, Violin JD, Que LG, Nelson CD, Benhar M, Keys JR, Rockman HA, Koch WJ, Daaka Y, Lefkowitz RJ, Stamler JS. Regulation of beta-adrenergic receptor signaling by S-nitrosylation of G-protein-coupled receptor kinase 2.Cell. 2007; 129:511–522. doi: 10.1016/j.cell.2007.02.046.CrossrefMedlineGoogle Scholar3. Huang ZM, Gao E, Fonseca FV, Hayashi H, Shang X, Hoffman NE, Chuprun JK, Tian X, Tilley DG, Madesh M, Lefer DJ, Stamler JS, Koch WJ. Convergence of G protein-coupled receptor and S-nitrosylation signaling determines the outcome to cardiac ischemic injury.Sci Signal. 2013; 6:ra95. doi: 10.1126/scisignal.2004225.CrossrefMedlineGoogle Scholar4. Choi H, Tostes RC, Webb RC. S-nitrosylation Inhibits protein kinase C-mediated contraction in mouse aorta.J Cardiovasc Pharmacol. 2011; 57:65–71. doi: 10.1097/FJC.0b013e3181fef9cb.CrossrefMedlineGoogle Scholar5. Yasukawa T, Tokunaga E, Ota H, Sugita H, Martyn JA, Kaneki M. S-nitrosylation-dependent inactivation of Akt/protein kinase B in insulin resistance.J Biol Chem. 2005; 280:7511–7518. doi: 10.1074/jbc.M411871200.CrossrefMedlineGoogle Scholar6. Reynaert NL, Ckless K, Korn SH, Vos N, Guala AS, Wouters EF, van der Vliet A, Janssen-Heininger YM. Nitric oxide represses inhibitory kappaB kinase through S-nitrosylation.Proc Natl Acad Sci USA. 2004; 101:8945–8950. doi: 10.1073/pnas.0400588101.CrossrefMedlineGoogle Scholar7. Coultrap SJ, Bayer KU. Nitric oxide induces Ca2+-independent activity of the Ca2+/calmodulin-dependent protein kinase II (CaMKII).J Biol Chem. 2014; 289:19458–19465. doi: 10.1074/jbc.M114.558254.CrossrefMedlineGoogle Scholar8. Park HS, Yu JW, Cho JH, Kim MS, Huh SH, Ryoo K, Choi EJ. Inhibition of apoptosis signal-regulating kinase 1 by nitric oxide through a thiol redox mechanism.J Biol Chem. 2004; 279:7584–7590. doi: 10.1074/jbc.M304183200.CrossrefMedlineGoogle Scholar9. Park HS, Huh SH, Kim MS, Lee SH, Choi EJ. Nitric oxide negatively regulates c-Jun N-terminal kinase/stress-activated protein kinase by means of S-nitrosylation.Proc Natl Acad Sci USA. 2000; 97:14382–14387. doi: 10.1073/pnas.97.26.14382.CrossrefMedlineGoogle Scholar10. Ozawa K, Whalen EJ, Nelson CD, Mu Y, Hess DT, Lefkowitz RJ, Stamler JS. S-nitrosylation of beta-arrestin regulates beta-adrenergic receptor trafficking.Mol Cell. 2008; 31:395–405. doi: 10.1016/j.molcel.2008.05.024.CrossrefMedlineGoogle Scholar11. Seth D, Hess DT, Hausladen A, Wang L, Wang YJ, Stamler JS. A multiplex enzymatic machinery for cellular protein s-nitrosylation.Mol Cell. 2018; 69:451–464.e6. doi: 10.1016/j.molcel.2017.12.025.CrossrefMedlineGoogle Scholar12. Kornberg MD, Sen N, Hara MR, Juluri KR, Nguyen JV, Snowman AM, Law L, Hester LD, Snyder SH. GAPDH mediates nitrosylation of nuclear proteins.Nat Cell Biol. 2010; 12:1094–1100. doi: 10.1038/ncb2114.CrossrefMedlineGoogle Scholar13. Cho DH, Nakamura T, Fang J, Cieplak P, Godzik A, Gu Z, Lipton SA. S-nitrosylation of Drp1 mediates beta-amyloid-related mitochondrial fission and neuronal injury.Science. 2009; 324:102–105. doi: 10.1126/science.1171091.CrossrefMedlineGoogle Scholar14. Jahani-Asl A, Huang E, Irrcher I, Rashidian J, Ishihara N, Lagace DC, Slack RS, Park DS. CDK5 phosphorylates DRP1 and drives mitochondrial defects in NMDA-induced neuronal death.Hum Mol Genet. 2015; 24:4573–4583. doi: 10.1093/hmg/ddv188.CrossrefMedlineGoogle Scholar15. Qu J, Nakamura T, Cao G, Holland EA, McKercher SR, Lipton SA. S-nitrosylation activates Cdk5 and contributes to synaptic spine loss induced by beta-amyloid peptide.Proc Natl Acad Sci USA. 2011; 108:14330–14335. doi: 10.1073/pnas.1105172108.CrossrefMedlineGoogle Scholar Previous Back to top Next FiguresReferencesRelatedDetailsCited By Gupta K, Kaladhar V, Fitzpatrick T, Fernie A, Møller I and Loake G (2022) Nitric oxide regulation of plant metabolism, Molecular Plant, 10.1016/j.molp.2021.12.012, 15:2, (228-242), Online publication date: 1-Feb-2022. Ye H, Wu J, Liang Z, Zhang Y and Huang Z (2022) Protein S -Nitrosation: Biochemistry, Identification, Molecular Mechanisms, and Therapeutic Applications , Journal of Medicinal Chemistry, 10.1021/acs.jmedchem.1c02194, 65:8, (5902-5925), Online publication date: 28-Apr-2022. Hu Q, Shi J, Zhang J, Wang Y, Guo Y and Zhang Z (2021) Progress and Prospects of Regulatory Functions Mediated by Nitric Oxide on Immunity and Immunotherapy, Advanced Therapeutics, 10.1002/adtp.202100032, 4:8, (2100032), Online publication date: 1-Aug-2021. Mule S, Manchola N, de Oliveira G, Pereira M, Magalhães R, Teixeira A, Colli W, Alves M and Palmisano G (2021) Proteome-wide modulation of S-nitrosylation in Trypanosoma cruzi trypomastigotes upon interaction with the host extracellular matrix, Journal of Proteomics, 10.1016/j.jprot.2020.104020, 231, (104020), Online publication date: 1-Jan-2021. Majumdar U, Manivannan S, Basu M, Ueyama Y, Blaser M, Cameron E, McDermott M, Lincoln J, Cole S, Wood S, Aikawa E, Lilly B and Garg V (2021) Nitric oxide prevents aortic valve calcification by S-nitrosylation of USP9X to activate NOTCH signaling, Science Advances, 10.1126/sciadv.abe3706, 7:6, Online publication date: 5-Feb-2021. Rizza S and Filomeni G (2020) Exploiting S- nitrosylation for cancer therapy: facts and perspectives , Biochemical Journal, 10.1042/BCJ20200064, 477:19, (3649-3672), Online publication date: 16-Oct-2020. Rizza S, Giglio P, Faienza F and Filomeni G (2019) Therapeutic Aspects of Protein Denitrosylation Therapeutic Application of Nitric Oxide in Cancer and Inflammatory Disorders, 10.1016/B978-0-12-816545-4.00009-8, (173-189), . Morris G, Puri B, Olive L, Carvalho A, Berk M and Maes M (2019) Emerging role of innate B1 cells in the pathophysiology of autoimmune and neuroimmune diseases: Association with inflammation, oxidative and nitrosative stress and autoimmune responses, Pharmacological Research, 10.1016/j.phrs.2019.104408, 148, (104408), Online publication date: 1-Oct-2019. Smolinski M, Green S and Storey K (2019) Glucose‐6‐phosphate dehydrogenase is posttranslationally regulated in the larvae of the freeze‐tolerant gall fly, Eurosta solidaginis , in response to freezing , Archives of Insect Biochemistry and Physiology, 10.1002/arch.21618, 102:4, Online publication date: 1-Dec-2019. Reis A, Stern A and Monteiro H (2019) S-nitrosothiols and H2S donors: Potential chemo-therapeutic agents in cancer, Redox Biology, 10.1016/j.redox.2019.101190, 27, (101190), Online publication date: 1-Oct-2019. Kayki-Mutlu G and Koch W (2021) Nitric Oxide and S-Nitrosylation in Cardiac Regulation: G Protein-Coupled Receptor Kinase-2 and β-Arrestins as Targets, International Journal of Molecular Sciences, 10.3390/ijms22020521, 22:2, (521) Shi X and Qiu H (2020) Post-Translational S-Nitrosylation of Proteins in Regulating Cardiac Oxidative Stress, Antioxidants, 10.3390/antiox9111051, 9:11, (1051) Sandalio L, Gotor C, Romero L and Romero-Puertas M (2019) Multilevel Regulation of Peroxisomal Proteome by Post-Translational Modifications, International Journal of Molecular Sciences, 10.3390/ijms20194881, 20:19, (4881) May 25, 2018Vol 122, Issue 11 Advertisement Article InformationMetrics © 2018 American Heart Association, Inc.https://doi.org/10.1161/CIRCRESAHA.118.313109PMID: 29798895 Originally publishedMay 25, 2018 Keywordsphosphorylationnitric oxidekinaseEditorialsPDF download Advertisement