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The Tumor Suppressor Sirt2 Regulates Cell Cycle Progression and Genome Stability by Modulating the Mitotic Deposition of H4K20 Methylation

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The Tumor Suppressor Sirt2 Regulates Cell Cycle Progression and Genome Stability by Modulating the Mitotic Deposition of H4K20 Methylation Downloaded from genesdev.cshlp.org on September 25, 2021 - Published by Cold Spring Harbor Laboratory Press The tumor suppressor SirT2 regulates cell cycle progression and genome stability by modulating the mitotic deposition of H4K20 methylation Lourdes Serrano,1,8,9 Paloma Martı´nez-Redondo,2,8 Anna Marazuela-Duque,2 Berta N. Vazquez,1 Scott J. Dooley,1 Philipp Voigt,3 David B. Beck,3 Noriko Kane-Goldsmith,1 Qiang Tong,4 Rosa M. Rabanal,5 Dolors Fondevila,5 Purificacio´ n Mun˜ oz,6 Marcus Kru¨ ger,7 Jay A. Tischfield,1 and Alejandro Vaquero2,9 1Department of Genetics, the Human Genetics Institute of New Jersey, Rutgers University, Piscataway, New Jersey 08854, USA; 2Chromatin Biology Laboratory, Cancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), 08908 L’Hospitalet de Llobregat, Barcelona, Spain; 3Department of Biochemistry, New York University School of Medicine, New York, New York 10016, USA; 4Children’s Nutrition Research Center, Baylor College of Medicine, Houston, Texas 77030, USA; 5Centre de Biotecnologia Animal i Tera`pia Ge`nica (CBATEG), Departament de Medicina i Cirurgia Animals, Universitat Auto` noma de Barcelona, 08193 Bellaterra (Cerdanyola del Valle`s), Spain; 6Ageing and Cancer Laboratory, Cancer Epigenetics and Biology Program (PEBC), Bellvitge Biomedical Research Institute (IDIBELL), 08908 L’Hospitalet de Llobregat, Barcelona, Spain; 7Max Planck Institute for Heart and Lung Research, Department of Cardiac Development and Remodeling, D-61231 Bad Nauheim, Germany The establishment of the epigenetic mark H4K20me1 (monomethylation of H4K20) by PR-Set7 during G2/M directly impacts S-phase progression and genome stability. However, the mechanisms involved in the regulation of this event are not well understood. Here we show that SirT2 regulates H4K20me1 deposition through the deacetylation of H4K16Ac (acetylation of H4K16) and determines the levels of H4K20me2/3 throughout the cell cycle. SirT2 binds and deacetylates PR-Set7 at K90, modulating its chromatin localization. Consistently, SirT2 depletion significantly reduces PR-Set7 chromatin levels, alters the size and number of PR-Set7 foci, and decreases the overall mitotic deposition of H4K20me1. Upon stress, the interaction between SirT2 and PR-Set7 increases along with the H4K20me1 levels, suggesting a novel mitotic checkpoint mechanism. SirT2 loss in mice induces significant defects associated with defective H4K20me1–3 levels. Accordingly, SirT2-deficient animals exhibit genomic instability and chromosomal aberrations and are prone to tumorigenesis. Our studies suggest that the dynamic cross-talk between the environment and the genome during mitosis determines the fate of the subsequent cell cycle. [Keywords: H4K20me1; PR-Set7; SIRT2; epigenetics; genome instability; sirtuins] Supplemental material is available for this article. Received November 30, 2012; revised version accepted February 11, 2013. The members of the Sir2 family of NAD+-dependent relevance of this activity for the Sir2 family remains deacetylases, also known as Sirtuins, are crucial factors a matter of debate (Tanny et al. 1999). Sirtuins participate in the response to metabolic, oxidative, or genotoxic stress in a myriad of functions directed at facilitating adapta- (Chalkiadaki and Guarente 2012). The capacity of Sirtuins tion to these stress conditions at both the cellular and to sense these compromising conditions appears to be due organismal levels. These functions include metabolic to the requirement of NAD+ for their enzymatic activity homeostasis, survival under stress, the maintenance of (Imai et al. 2000; Landry et al. 2000; Smith et al. 2000). A genomic stability, and cell differentiation and development second enzymatic activity, a mono-ADP ribosyltransferase (Saunders and Verdin 2007; Finkel et al. 2009). activity, has been reported for some Sirtuins, although the Sirtuins have been present since early in evolution. From this time, they have apparently played a major role in the cross-talk between environmental stimuli and geno- 8These authors contributed equally to this work. 9Corresponding authors mic information (Vaquero 2009). In fact, eukaryotic Sirtuins E-mail [email protected] often exert their functions by affecting chromatin. These E-mail [email protected] Article published online ahead of print. Article and publication date are functions include gene silencing, chromatin structure online at http://www.genesdev.org/cgi/doi/10.1101/gad.211342.112. modulation, DNA damage signaling, DNA repair, and cell GENES & DEVELOPMENT 27:639–653 Ó 2013 by Cold Spring Harbor Laboratory Press ISSN 0890-9369/13; www.genesdev.org 639 Downloaded from genesdev.cshlp.org on September 25, 2021 - Published by Cold Spring Harbor Laboratory Press Serrano et al. cycle regulation. The mechanisms through which Sirtuins effects of SirT2 on chromatin appear to be restricted to act on chromatin involve the deacetylation of both histone early mitosis, as SirT2 dissociates from the chromosomes and nonhistone proteins ranging from chromatin machin- during metaphase (Vaquero et al. 2006). ery enzymes to transcription factors. Several levels of interplay have been identified between SirT2 is among the least understood of the seven H4K16Ac and other histone marks. The best characterized mammalian Sirtuins. To date, it has been linked to the of these are linked to the transcriptional role of H4K16Ac regulation of mitotic progression (Dryden et al. 2003), at the level of initiation (H3K4me3 and H3S10P) or elonga- oxidative stress response (Wang et al. 2007), metabolism tion (H3K36me3) (Dou et al. 2005; Bell et al. 2007; Zippo (Jiang et al. 2011), microtubule dynamics (North et al. et al. 2009). Interestingly, an antagonism between H4K16Ac 2003), cell migration (Pandithage et al. 2008), apoptosis and H4K20 methylation has been proposed. In vitro stud- (Li et al. 2011), neurotoxicity (Outeiro et al. 2007), and the ies with peptides have shown that H4K16Ac could in- inhibition of differentiation (Jing et al. 2007; Li et al. hibit the monomethylation of H4K20 (H4K20me1) by the 2007). The role of SirT2 in cell cycle regulation is par- histone methyltransferase (HMT) PR-Set7 (also known as ticularly intriguing. SirT2 has been found to be up-regulated SET8 or KMT5A) (Nishioka et al. 2002a). The relevance of during mitosis, and its overexpression has been reported this antagonism lies in the importance of H4K20 mono-, to induce the lengthening of mitosis (Dryden et al. 2003) di-, or trimethylation in the control of cell cycle progres- and shortening of G1 (Bae et al. 2004). Additionally, micro- sion, chromosome compaction, development, DNA re- injection of either SirT2 or its yeast ortholog, Hst2p, has pair signaling, and genome stability (Sanders et al. 2004; been shown to inhibit starfish oocyte maturation and Schotta et al. 2004, 2008; Karachentsev et al. 2005; Oda embryonic cell division (Borra et al. 2002). Primary mouse et al. 2009). H4K20me1 is established in late G2/early M embryonic fibroblasts (MEFs) derived from SirT2 knock- by PR-Set7 and is paramount in metaphasic chromosome out mice show a very limited decrease in the length of compaction and mitotic exit during mitosis (Houston mitosis but longer G1 and shorter S phases—a finding that et al. 2008; Oda et al. 2009) and in DNA repair and DNA confirms that SirT2 has a significant role in the cell cycle replication (Jorgensen et al. 2007; Tardat et al. 2007, and, at least in part, a direct impact on G1/S (Vaquero 2010). During late M/early G1, some H4K20me1 is further et al. 2006). Recent studies have suggested a role for SirT2 methylated into H4K20me2 (required for DNA repair) or in the control of mitotic exit by acting on the anaphase- H4K20me3 (required for heterochromatin structure for- promoting complex; namely, via the deacetylation of its mation) by the HMTs Suv4-20h1 or Suv4-20h2, respec- coactivators, APCCDH1 and CDC20 (Kim et al. 2011). tively (Sanders et al. 2004; Schotta et al. 2004, 2008). PR- A very interesting link between chromatin regulation Set7 levels are tightly regulated during the cell cycle, and and SirT2 that directly points to cell cycle regulation is this protein is clearly detected from late M to early G1 (Oda the acetylation of Lys 16 on histone H4 (H4K16Ac), a et al. 2009). Despite the importance of H4K20 methylation mark that has been proposed to directly regulate chro- to cell cycle progression, little is known about the signals matin structure (Shia et al. 2006). Previous studies have and pathways that regulate this modification or the mech- determined that in terms of deacetylase activity, SirT2 anisms through which the enzymes involved are modu- is highly specific for H4K16Ac (Vaquero et al. 2006), in lated. The only clue currently is the reported phosphor- contrast to the other major mammalian H4K16Ac de- ylation of S29 in PR-Set7 by CDK1, which appears to be acetylase, the Sirtuin SirT1, which has a wider specificity important for regulating the binding of PR-Set7 to chro- (Vaquero et al. 2004). Studies have shown that H4K16Ac matin and for preventing its degradation by APCCDH1- inhibits the folding of chromatin fiber in vitro and there- mediated polyubiquitination (Wu et al. 2010). fore facilitates the formation of higher orders of chromatin WhileinvestigatingtheroleofSirT2andH4K16Acin organization (Shogren-Knaak et al. 2006; Robinson et al. cell cycle control and genome stability, we discovered roles 2008). The acetylation/deacetylation of H4K16 has been
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