Proliferating cell nuclear antigen acts as a cytoplasmic platform controlling human neutrophil survival
Véronique Witko-Sarsat, Julie Mocek, Dikra Bouayad, Nicola Tamassia, Jean-Antoine Ribeil, Céline Candalh, Noélie Davezac, Nathalie Reuter, Luc Mouthon, Olivier Hermine, Magali Pederzoli-Ribeil, Marco A Cassatella, Véronique Witko-Sarsat, Julie Mocek, Dikra Bouayad, Nicola Tamassia, Jean-Antoine Ribeil, Céline Candalh, Noélie Davezac, Nathalie Reuter, Luc Mouthon, Olivier Hermine, Magali Pederzoli-Ribeil, Marco A Cassatella
Abstract
Neutrophil apoptosis is a highly regulated process essential for inflammation resolution, the molecular mechanisms of which are only partially elucidated. In this study, we describe a survival pathway controlled by proliferating cell nuclear antigen (PCNA), a nuclear factor involved in DNA replication and repairing of proliferating cells. We show that mature neutrophils, despite their inability to proliferate, express high levels of PCNA exclusively in their cytosol and constitutively associated with procaspases, presumably to prevent their activation. Notably, cytosolic PCNA abundance decreased during apoptosis, and increased during in vitro and in vivo exposure to the survival factor granulocyte colony-stimulating factor (G-CSF). Peptides derived from the cyclin-dependent kinase inhibitor p21, which compete with procaspases to bind PCNA, triggered neutrophil apoptosis thus demonstrating that specific modification of PCNA protein interactions affects neutrophil survival. Furthermore, PCNA overexpression rendered neutrophil-differentiated PLB985 myeloid cells significantly more resistant to TNF-related apoptosis-inducing ligand- or gliotoxin-induced apoptosis. Conversely, a decrease in PCNA expression after PCNA small interfering RNA transfection sensitized these cells to apoptosis. Finally, a mutation in the PCNA interdomain-connecting loop, the binding site for many partners, significantly decreased the PCNA-mediated antiapoptotic effect. These results identify PCNA as a regulator of neutrophil lifespan, thereby highlighting a novel target to potentially modulate pathological inflammation.
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References
- Altznauer F., Martinelli S., Yousefi S., Thürig C., Schmid I., Conway E.M., Schöni M.H., Vogt P., Mueller C., Fey M.F., et al. 2004. Inflammation-associated cell cycle–independent block of apoptosis by survivin in terminally differentiated neutrophils. J. Exp. Med. 199:1343–1354 10.1084/jem.20032033
- Basu S., Hodgson G., Katz M., Dunn A.R. 2002. Evaluation of role of G-CSF in the production, survival, and release of neutrophils from bone marrow into circulation. Blood. 100:854–861 10.1182/blood.V100.3.854
- Borregaard N., Heiple J.M., Simons E.R., Clark R.A. 1983. Subcellular localization of the b-cytochrome component of the human neutrophil microbicidal oxidase: translocation during activation. J. Cell Biol. 97:52–61 10.1083/jcb.97.1.52
- Cassatella M.A. 1999. Neutrophil-derived proteins: selling cytokines by the pound. Adv. Immunol. 73:369–509 10.1016/S0065-2776(08)60791-9
- Cowland J.B., Borregaard N. 1999. Isolation of neutrophil precursors from bone marrow for biochemical and transcriptional analysis. J. Immunol. Methods. 232:191–200 10.1016/S0022-1759(99)00176-3
- Derouet M., Thomas L., Cross A., Moots R.J., Edwards S.W. 2004. Granulocyte macrophage colony-stimulating factor signaling and proteasome inhibition delay neutrophil apoptosis by increasing the stability of Mcl-1. J. Biol. Chem. 279:26915–26921 10.1074/jbc.M313875200
- Dong C., Lyu S.C., Krensky A.M., Clayberger C. 2003. DQ 65-79, a peptide derived from HLA class II, mimics p21 to block T cell proliferation. J. Immunol. 171:5064–5070
- Drewniak A., van Raam B.J., Geissler J., Tool A.T., Mook O.R., van den Berg T.K., Baas F., Kuijpers T.W. 2009. Changes in gene expression of granulocytes during in vivo granulocyte colony-stimulating factor/dexamethasone mobilization for transfusion purposes. Blood. 113:5979–5998 10.1182/blood-2008-10-182147
- Fox S., Leitch A.E., Duffin R., Haslett C., Rossi A.G. 2010. Neutrophil apoptosis: relevance to the innate immune response and inflammatory disease. J. Innate Immun. 2:216–227 10.1159/000284367
- Fujise K., Zhang D., Liu J., Yeh E.T. 2000. Regulation of apoptosis and cell cycle progression by MCL1. Differential role of proliferating cell nuclear antigen. J. Biol. Chem. 275:39458–39465 10.1074/jbc.M006626200
- Goepel F., Weinmann P., Schymeinsky J., Walzog B. 2004. Identification of caspase-10 in human neutrophils and its role in spontaneous apoptosis. J. Leukoc. Biol. 75:836–843 10.1189/jlb.0703317
- Hajjar E., Korkmaz B., Gauthier F., Brandsdal B.O., Witko-Sarsat V., Reuter N. 2006. Inspection of the binding sites of proteinase3 for the design of a highly specific substrate. J. Med. Chem. 49:1248–1260 10.1021/jm051018t
- Hallett J.M., Leitch A.E., Riley N.A., Duffin R., Haslett C., Rossi A.G. 2008. Novel pharmacological strategies for driving inflammatory cell apoptosis and enhancing the resolution of inflammation. Trends Pharmacol. Sci. 29:250–257 10.1016/j.tips.2008.03.002
- Henson P.M. 2005. Dampening inflammation. Nat. Immunol. 6:1179–1181 10.1038/ni1205-1179
- Hino M., Suzuki K., Yamane T., Sakai N., Kubota H., Koh K.R., Ohta K., Hato F., Kitagawa S., Tatsumi N. 2000. Ex vivo expansion of mature human neutrophils with normal functions from purified peripheral blood CD34+ haematopoietic progenitor cells. Br. J. Haematol. 109:314–321 10.1046/j.1365-2141.2000.02054.x
- Kantari C., Pederzoli-Ribeil M., Amir-Moazami O., Gausson-Dorey V., Moura I.C., Lecomte M.C., Benhamou M., Witko-Sarsat V. 2007. Proteinase 3, the Wegener autoantigen, is externalized during neutrophil apoptosis: evidence for a functional association with phospholipid scramblase 1 and interference with macrophage phagocytosis. Blood. 110:4086–4095 10.1182/blood-2007-03-080457
- Klausen P., Bjerregaard M.D., Borregaard N., Cowland J.B. 2004. End-stage differentiation of neutrophil granulocytes in vivo is accompanied by up-regulation of p27kip1 and down-regulation of CDK2, CDK4, and CDK6. J. Leukoc. Biol. 75:569–578 10.1189/jlb.1003474
- Koedel U., Frankenberg T., Kirschnek S., Obermaier B., Häcker H., Paul R., Häcker G. 2009. Apoptosis is essential for neutrophil functional shutdown and determines tissue damage in experimental pneumococcal meningitis. PLoS Pathog. 5:e1000461 10.1371/journal.ppat.1000461
- Koundrioukoff S., Jónsson Z.O., Hasan S., de Jong R.N., van der Vliet P.C., Hottiger M.O., Hübscher U. 2000. A direct interaction between proliferating cell nuclear antigen (PCNA) and Cdk2 targets PCNA-interacting proteins for phosphorylation. J. Biol. Chem. 275:22882–22887 10.1074/jbc.M001850200
- Leavitt R.Y., Fauci A.S., Bloch D.A., Michel B.A., Hunder G.G., Arend W.P., Calabrese L.H., Fries J.F., Lie J.T., Lightfoot R.W., Jr, et al. 1990. The American College of Rheumatology 1990 criteria for the classification of Wegener’s granulomatosis. Arthritis Rheum. 33:1101–1107 10.1002/art.1780330807
- Luqmani R.A., Bacon P.A., Moots R.J., Janssen B.A., Pall A., Emery P., Savage C., Adu D. 1994. Birmingham Vasculitis Activity Score (BVAS) in systemic necrotizing vasculitis. QJM. 87:671–678
- Maga G., Hubscher U. 2003. Proliferating cell nuclear antigen (PCNA): a dancer with many partners. J. Cell Sci. 116:3051–3060 10.1242/jcs.00653
- Maianski N.A., Roos D., Kuijpers T.W. 2004. Bid truncation, bid/bax targeting to the mitochondria, and caspase activation associated with neutrophil apoptosis are inhibited by granulocyte colony-stimulating factor. J. Immunol. 172:7024–7030
- Mattock H., Jares P., Zheleva D.I., Lane D.P., Warbrick E., Blow J.J. 2001a. Use of peptides from p21 (Waf1/Cip1) to investigate PCNA function in Xenopus egg extracts. Exp. Cell Res. 265:242–251 10.1006/excr.2001.5181
- Mattock H., Lane D.P., Warbrick E. 2001b. Inhibition of cell proliferation by the PCNA-binding region of p21 expressed as a GFP miniprotein. Exp. Cell Res. 265:234–241 10.1006/excr.2001.5160
- McStay G.P., Salvesen G.S., Green D.R. 2008. Overlapping cleavage motif selectivity of caspases: implications for analysis of apoptotic pathways. Cell Death Differ. 15:322–331 10.1038/sj.cdd.4402260
- Mócsai A., Zhou M., Meng F., Tybulewicz V.L., Lowell C.A. 2002. Syk is required for integrin signaling in neutrophils. Immunity. 16:547–558 10.1016/S1074-7613(02)00303-5
- Moldovan G.L., Pfander B., Jentsch S. 2007. PCNA, the maestro of the replication fork. Cell. 129:665–679 10.1016/j.cell.2007.05.003
- Moriceau S., Kantari C., Mocek J., Davezac N., Gabillet J., Guerrera I.C., Brouillard F., Tondelier D., Sermet-Gaudelus I., Danel C., et al. 2009. Coronin-1 is associated with neutrophil survival and is cleaved during apoptosis: potential implication in neutrophils from cystic fibrosis patients. J. Immunol. 182:7254–7263 10.4049/jimmunol.0803312
- Moriceau S., Lenoir G., Witko-Sarsat V. 2010. In cystic fibrosis homozygotes and heterozygotes, neutrophil apoptosis is delayed and modulated by diamide or roscovitine: evidence for an innate neutrophil disturbance. J. Innate Immun. 2:260–266 10.1159/000295791
- Murphy B.M., O’Neill A.J., Adrain C., Watson R.W., Martin S.J. 2003. The apoptosome pathway to caspase activation in primary human neutrophils exhibits dramatically reduced requirements for cytochrome c. J. Exp. Med. 197:625–632 10.1084/jem.20021862
- Naryzhny S.N., Lee H. 2004. The post-translational modifications of proliferating cell nuclear antigen: acetylation, not phosphorylation, plays an important role in the regulation of its function. J. Biol. Chem. 279:20194–20199 10.1074/jbc.M312850200
- Nathan C. 2006. Neutrophils and immunity: challenges and opportunities. Nat. Rev. Immunol. 6:173–182 10.1038/nri1785
- Pedruzzi E., Fay M., Elbim C., Gaudry M., Gougerot-Pocidalo M.A. 2002. Differentiation of PLB-985 myeloid cells into mature neutrophils, shown by degranulation of terminally differentiated compartments in response to N-formyl peptide and priming of superoxide anion production by granulocyte-macrophage colony-stimulating factor. Br. J. Haematol. 117:719–726 10.1046/j.1365-2141.2002.03521.x
- Roa S., Avdievich E., Peled J.U., Maccarthy T., Werling U., Kuang F.L., Kan R., Zhao C., Bergman A., Cohen P.E., et al. 2008. Ubiquitylated PCNA plays a role in somatic hypermutation and class-switch recombination and is required for meiotic progression. Proc. Natl. Acad. Sci. USA. 105:16248–16253 10.1073/pnas.0808182105
- Rossato M., Cencig S., Gasperini S., Cassatella M.A., Bazzoni F. 2007. IL-10 modulates cytokine gene transcription by protein synthesis-independent and dependent mechanisms in lipopolysaccharide-treated neutrophils. Eur. J. Immunol. 37:3176–3189 10.1002/eji.200737625
- Rossi A.G., Sawatzky D.A., Walker A., Ward C., Sheldrake T.A., Riley N.A., Caldicott A., Martinez-Losa M., Walker T.R., Duffin R., et al. 2006. Cyclin-dependent kinase inhibitors enhance the resolution of inflammation by promoting inflammatory cell apoptosis. Nat. Med. 12:1056–1064 10.1038/nm1468
- Scapini P., Nardelli B., Nadali G., Calzetti F., Pizzolo G., Montecucco C., Cassatella M.A. 2003. G-CSF–stimulated neutrophils are a prominent source of functional BLyS. J. Exp. Med. 197:297–302 10.1084/jem.20021343
- Serhan C.N., Savill J. 2005. Resolution of inflammation: the beginning programs the end. Nat. Immunol. 6:1191–1197 10.1038/ni1276
- Simon H.U. 2003. Neutrophil apoptosis pathways and their modifications in inflammation. Immunol. Rev. 193:101–110 10.1034/j.1600-065X.2003.00038.x
- Tamassia N., Le Moigne V., Rossato M., Donini M., McCartney S., Calzetti F., Colonna M., Bazzoni F., Cassatella M.A. 2008. Activation of an immunoregulatory and antiviral gene expression program in poly(I:C)-transfected human neutrophils. J. Immunol. 181:6563–6573
- Theilgaard-Mönch K., Porse B.T., Borregaard N. 2006. Systems biology of neutrophil differentiation and immune response. Curr. Opin. Immunol. 18:54–60 10.1016/j.coi.2005.11.010
- Waga S., Hannon G.J., Beach D., Stillman B. 1994. The p21 inhibitor of cyclin-dependent kinases controls DNA replication by interaction with PCNA. Nature. 369:574–578 10.1038/369574a0
- Warbrick E. 2000. The puzzle of PCNA’s many partners. Bioessays. 22:997–1006 10.1002/1521-1878(200011)22:11<997::AID-BIES6>;2-#
- Warbrick E. 2006. A functional analysis of PCNA-binding peptides derived from protein sequence, interaction screening and rational design. Oncogene. 25:2850–2859 10.1038/sj.onc.1209320
- Ward C., Chilvers E.R., Lawson M.F., Pryde J.G., Fujihara S., Farrow S.N., Haslett C., Rossi A.G. 1999. NF-kappaB activation is a critical regulator of human granulocyte apoptosis in vitro. J. Biol. Chem. 274:4309–4318 10.1074/jbc.274.7.4309
- Witko-Sarsat V., Rieu P., Descamps-Latscha B., Lesavre P., Halbwachs-Mecarelli L. 2000. Neutrophils: molecules, functions and pathophysiological aspects. Lab. Invest. 80:617–653
- Yin W., Raffelsberger W., Gronemeyer H. 2005. Retinoic acid determines life span of leukemic cells by inducing antagonistic apoptosis-regulatory programs. Int. J. Biochem. Cell Biol. 37:1696–1708 10.1016/j.biocel.2005.03.003
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