Abstract
Senescent cells exhibit dramatic changes in protein post-translational modifications. Here, we describe a method, stable isotope labeling with amino acids in cell culture (SILAC) coupled to liquid chromatography tandem mass spectrometry (LC-MS/MS), to identify changes in the ubiquitinome in cells that have undergone oncogene-induced senescence.
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References
Munoz-Espin D, Canamero M, Maraver A, Gomez-Lopez G, Contreras J, Murillo-Cuesta S et al (2013) Programmed cell senescence during mammalian embryonic development. Cell 155:1104–1118
Rodier F, Campisi J (2011) Four faces of cellular senescence. J Cell Biol 192:547–556
Collado M, Serrano M (2010) Senescence in tumours: evidence from mice and humans. Nat Rev Cancer 10:51–57
Hayflick L, Moorhead PS (1961) The serial cultivation of human diploid cell strains. Exp Cell Res 585–621
Harley CB, Futcher AB, Greider CW (1990) Telomeres shorten during ageing of human fibroblasts. Nature 345:458–460
Yang J, Chang E, Cherry AM, Bangs CD, Oei Y, Bodnar A et al (1999) Human endothelial cell life extension by telomerase expression. J Biol Chem 274:26141–26148
Chen Z, Trotman LC, Shaffer D, Lin HK, Dotan ZA, Niki M (2005) Crucial role of p53-dependent cellular senescence in suppression of Pten-deficient tumorigenesis. Nature 436:725–730
Yaswen P, Campisi J (2007) Oncogene-induced senescence pathways weave an intricate tapestry. Cell 128:233–234
Serrano M, Lin AW, McCurrach ME, Beach D, Lowe S (1997) Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a. Cell 88:593–602
Li M, Durbin KR, Sweet SM, Tipton JD, Zheng Y, Kelleher NL (2013) Oncogene-induced cellular senescence elicits an anti-Warburg effect. Proteomics 13:2585–2596
Nalepa G, Rolfe M, Harper JW (2006) Drug discovery in the ubiquitin-proteasome system. Nat Rev Drug Discov 5:596–613
Chau V, Tobias JW, Bachmair A, Marriott D, Ecker DJ, Gonda DK, Varshavsky A (1989) A multiubiquitin chain is confined to a specific lysine in a targeted short-lived protein. Science 243:1576–1583
Neutzner M, Neutzner A (2012) Enzymes of ubiquitination and deubiquitination. Essays Biochem 52:37–50
Hershko A, Ciechanover A, Varshavsky A (2000) Basic medical research award. The ubiquitin system. Nat Med 6:1073–1081
Pickart CM (2001) Mechanisms underlying ubiquitination. Annu Rev Biochem 70:503–533
Johmura Y, Sun J, Kitagawa K, Nakanishi K, Kuno T, Naiki-Ito A et al (2016) SCF(Fbxo22)-KDM4A targets methylated p53 for degradation and regulates senescence. Nat Commun 7:10574
Zhu H, Ren S, Bitler BG, Aird KM, Tu Z, Skordalakes E et al (2015) SPOP E3 ubiquitin ligase adaptor promotes cellular senescence by degrading the SENP7 deSUMOylase. Cell Rep 13:1183–1193
Cox J, Mann M (2008) MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. Nat Biotechnol 26:1367–1372
Acknowledgment
This work was supported in part by NIH/NCI grants R01CA160331 to R.Z., R01CA131582 to D.W.S., and an institutional grant to The Wistar Institute (NCI Cancer Core Grant CA010815).
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Zhu, H., Le, L., Tang, HY., Speicher, D.W., Zhang, R. (2017). Detection of the Ubiquitinome in Cells Undergoing Oncogene-Induced Senescence. In: Nikiforov, M. (eds) Oncogene-Induced Senescence. Methods in Molecular Biology, vol 1534. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6670-7_12
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DOI: https://doi.org/10.1007/978-1-4939-6670-7_12
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Online ISBN: 978-1-4939-6670-7
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