ABSTRACT In addition to allelic mutations, cancers are known to harbor alterations in their chromatin landscape. Here we show that genomic ablation of Smad ubiquitin regulatory factor 2

(Smurf2), a HECT-domain E3 ubiquitin ligase, results in dysregulation of both the DNA damage response and genomic stability, culminating in increased susceptibility to various types of

cancers in aged mice. We show that Smurf2 regulates the monoubiquitination of histone H2B as well as the trimethylation of histone H3 at Lys4 and Lys79 by targeting ring finger protein 20

(RNF20) for proteasomal degradation in both mouse and human cells. We also show that Smurf2 and RNF20 are colocalized at the γ-H2AX foci of double-stranded DNA breaks in the nucleus. Thus,

issues and online access $209.00 per year only $17.42 per issue Learn more Buy this article * Purchase on SpringerLink * Instant access to full article PDF Buy now Prices may be subject to

local taxes which are calculated during checkout ADDITIONAL ACCESS OPTIONS: * Log in * Learn about institutional subscriptions * Read our FAQs * Contact customer support SIMILAR CONTENT

BEING VIEWED BY OTHERS TARGETING DCAF5 SUPPRESSES _SMARCB1_-MUTANT CANCER BY STABILIZING SWI/SNF Article 27 March 2024 THE SWI/SNF ATPASE BRG1 FACILITATES MULTIPLE PRO-TUMORIGENIC GENE

EXPRESSION PROGRAMS IN _SMARCB1_-DEFICIENT CANCER CELLS Article Open access 01 June 2022 THE ASSEMBLY OF MAMMALIAN SWI/SNF CHROMATIN REMODELING COMPLEXES IS REGULATED BY LYSINE-METHYLATION

DEPENDENT PROTEOLYSIS Article Open access 05 November 2022 REFERENCES * Hershko, A. & Ciechanover, A. The ubiquitin system. _Annu. Rev. Biochem._ 67, 425–479

10.1146/annurev.biochem.67.1.425 (1998). Article  CAS  Google Scholar  * Izzi, L. & Attisano, L. Regulation of the TGF-β signalling pathway by ubiquitin-mediated degradation. _Oncogene_

23, 2071–2078 (2004). Article  CAS  Google Scholar  * Lönn, P., Moren, A., Raja, E., Dahl, M. & Moustakas, A. Regulating the stability of TGF-β receptors and Smads. _Cell Res._ 19, 21–35

(2009). Article  Google Scholar  * Zhang, Y., Chang, C., Gehling, D.J., Hemmati-Brivanlou, A. & Derynck, R. Regulation of Smad degradation and activity by Smurf2, an E3 ubiquitin

ligase. _Proc. Natl. Acad. Sci. USA_ 98, 974–979 (2001). Article  CAS  Google Scholar  * Lin, X., Liang, M. & Feng, X.H. Smurf2 is a ubiquitin E3 ligase mediating proteasome-dependent

degradation of Smad2 in transforming growth factor-β signaling. _J. Biol. Chem._ 275, 36818–36822 (2000). Article  CAS  Google Scholar  * Kavsak, P. et al. Smad7 binds to Smurf2 to form an

E3 ubiquitin ligase that targets the TGF-β receptor for degradation. _Mol. Cell_ 6, 1365–1375 (2000). Article  CAS  Google Scholar  * Li, H. & Seth, A. An RNF11: Smurf2 complex mediates

ubiquitination of the AMSH protein. _Oncogene_ 23, 1801–1808 (2004). Article  CAS  Google Scholar  * Schwamborn, J.C., Muller, M., Becker, A.H. & Puschel, A.W. Ubiquitination of the

GTPase Rap1B by the ubiquitin ligase Smurf2 is required for the establishment of neuronal polarity. _EMBO J._ 26, 1410–1422 (2007). Article  CAS  Google Scholar  * Narimatsu, M. et al.

Regulation of planar cell polarity by Smurf ubiquitin ligases. _Cell_ 137, 295–307 (2009). Article  CAS  Google Scholar  * Zhang, H. & Cohen, S.N. Smurf2 up-regulation activates

telomere-dependent senescence. _Genes Dev._ 18, 3028–3040 (2004). Article  CAS  Google Scholar  * Fukuchi, M. et al. High-level expression of the Smad ubiquitin ligase Smurf2 correlates with

poor prognosis in patients with esophageal squamous cell carcinoma. _Cancer Res._ 62, 7162–7165 (2002). CAS  PubMed  Google Scholar  * Jin, C. et al. Smad ubiquitination regulatory factor 2

promotes metastasis of breast cancer cells by enhancing migration and invasiveness. _Cancer Res._ 69, 735–740 (2009). Article  CAS  Google Scholar  * Johnstone, S.E. & Baylin, S.B.

Stress and the epigenetic landscape: a link to the pathobiology of human diseases? _Nat. Rev. Genet._ 11, 806–812 (2010). Article  CAS  Google Scholar  * Kim, J., Hake, S.B. & Roeder,

R.G. The human homolog of yeast BRE1 functions as a transcriptional coactivator through direct activator interactions. _Mol. Cell_ 20, 759–770 (2005). Article  CAS  Google Scholar  * Zhu, B.

et al. Monoubiquitination of human histone H2B: the factors involved and their roles in _HOX_ gene regulation. _Mol. Cell_ 20, 601–611 (2005). Article  CAS  Google Scholar  * Minsky, N. et

al. Monoubiquitinated H2B is associated with the transcribed region of highly expressed genes in human cells. _Nat. Cell Biol._ 10, 483–488 (2008). Article  CAS  Google Scholar  * Moyal, L.

et al. Requirement of ATM-dependent monoubiquitylation of histone H2B for timely repair of DNA double-strand breaks. _Mol. Cell_ 41, 529–542 (2011). Article  CAS  Google Scholar  * Nakamura,

K. et al. Regulation of homologous recombination by RNF20-dependent H2B ubiquitination. _Mol. Cell_ 41, 515–528 (2011). Article  CAS  Google Scholar  * Tang, L.Y. et al. Ablation of Smurf2

reveals an inhibition in TGF-β signalling through multiple mono-ubiquitination of Smad3. _EMBO J._ 30, 4777–4789 (2011). Article  CAS  Google Scholar  * Todaro, G.J. & Green, H.

Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. _J. Cell Biol._ 17, 299–313 (1963). Article  CAS  Google Scholar  * Harvey,

D.M. & Levine, A.J. p53 alteration is a common event in the spontaneous immortalization of primary BALB/c murine embryo fibroblasts. _Genes Dev._ 5, 2375–2385 (1991). Article  CAS 

Google Scholar  * Sherr, C.J. Tumor surveillance via the ARF-p53 pathway. _Genes Dev._ 12, 2984–2991 (1998). Article  CAS  Google Scholar  * Franken, N.A., Rodermond, H.M., Stap, J.,

Haveman, J. & van Bree, C. Clonogenic assay of cells _in vitro_. _Nat. Protoc._ 1, 2315–2319 (2006). Article  CAS  Google Scholar  * Montecucco, A. & Biamonti, G. Cellular response

to etoposide treatment. _Cancer Lett._ 252, 9–18 (2007). Article  CAS  Google Scholar  * Bonner, W.M. et al. γH2AX and cancer. _Nat. Rev. Cancer_ 8, 957–967 (2008). Article  CAS  Google

Scholar  * Falk, M., Lukasova, E. & Kozubek, S. Chromatin structure influences the sensitivity of DNA to gamma-radiation. _Biochim. Biophys. Acta_ 1783, 2398–2414 (2008). Article  CAS 

Google Scholar  * Murga, M. et al. Global chromatin compaction limits the strength of the DNA damage response. _J. Cell Biol._ 178, 1101–1108 (2007). Article  CAS  Google Scholar  *

Shilatifard, A. Chromatin modifications by methylation and ubiquitination: implications in the regulation of gene expression. _Annu. Rev. Biochem._ 75, 243–269 (2006). Article  CAS  Google

Scholar  * Weake, V.M. & Workman, J.L. Histone ubiquitination: triggering gene activity. _Mol. Cell_ 29, 653–663 (2008). Article  CAS  Google Scholar  * Sun, Z.W. & Allis, C.D.

Ubiquitination of histone H2B regulates H3 methylation and gene silencing in yeast. _Nature_ 418, 104–108 (2002). Article  CAS  Google Scholar  * Laribee, R.N. et al. BUR kinase selectively

regulates H3 K4 trimethylation and H2B ubiquitylation through recruitment of the PAF elongation complex. _Curr. Biol._ 15, 1487–1493 (2005). Article  CAS  Google Scholar  * Lee, J.S. et al.

Histone crosstalk between H2B monoubiquitination and H3 methylation mediated by COMPASS. _Cell_ 131, 1084–1096 (2007). Article  CAS  Google Scholar  * Kim, J. et al. RAD6-Mediated

transcription-coupled H2B ubiquitylation directly stimulates H3K4 methylation in human cells. _Cell_ 137, 459–471 (2009). Article  CAS  Google Scholar  * Kizer, K.O. et al. A novel domain in

Set2 mediates RNA polymerase II interaction and couples histone H3 K36 methylation with transcript elongation. _Mol. Cell. Biol._ 25, 3305–3316 (2005). Article  CAS  Google Scholar  *

Berglund, L. et al. A genecentric Human Protein Atlas for expression profiles based on antibodies. _Mol. Cell. Proteomics_ 7, 2019–2027 (2008). Article  CAS  Google Scholar  * Haber, D.

& Harlow, E. Tumour-suppressor genes: evolving definitions in the genomic age. _Nat. Genet._ 16, 320–322 (1997). Article  CAS  Google Scholar  * Fukunaga, E. et al. Smurf2 induces

ubiquitin-dependent degradation of Smurf1 to prevent migration of breast cancer cells. _J. Biol. Chem._ 283, 35660–35667 (2008). Article  CAS  Google Scholar  * Fierz, B. et al. Histone H2B

ubiquitylation disrupts local and higher-order chromatin compaction. _Nat. Chem. Biol._ 7, 113–119 (2011). Article  CAS  Google Scholar  * Hogan, B., Beddington, R., Constantini, F. &

Lacy, E. Manipulating the Mouse Embryo: A Laboratory Manual. 2 edn. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA, 1994). * Blank, M., Lerenthal, Y., Mittelman, L.

& Shiloh, Y. Condensin I recruitment and uneven chromatin condensation precede mitotic cell death in response to DNA damage. _J. Cell Biol._ 174, 195–206 (2006). Article  CAS  Google

Scholar  * Galanty, Y. et al. Mammalian SUMO E3-ligases PIAS1 and PIAS4 promote responses to DNA double-strand breaks. _Nature_ 462, 935–939 (2009). Article  CAS  Google Scholar  * Rudolph,

C., Adam, G. & Simm, A. Determination of copy number of c-Myc protein per cell by quantitative Western blotting. _Anal. Biochem._ 269, 66–71 (1999). Article  CAS  Google Scholar  *

Shechter, D., Dormann, H.L., Allis, C.D. & Hake, S.B. Extraction, purification and analysis of histones. _Nat. Protoc._ 2, 1445–1457 (2007). Article  CAS  Google Scholar  * Ziv, Y. et

al. Chromatin relaxation in response to DNA double-strand breaks is modulated by a novel ATM- and KAP-1 dependent pathway. _Nat. Cell Biol._ 8, 870–876 (2006). Article  CAS  Google Scholar 

* Zaret, K. Micrococcal nuclease analysis of chromatin structure. _Curr Protoc Mol. Biol._ Chapter 21, Unit 21 (2005). * Yamashita, M. et al. Ubiquitin ligase Smurf1 controls osteoblast

activity and bone homeostasis by targeting MEKK2 for degradation. _Cell_ 121, 101–113 (2005). Article  CAS  Google Scholar  Download references ACKNOWLEDGEMENTS We thank M. Anver for

pathology services, V. Barr for assistance with microscope, X. Wu for assistance with the microarray experiments, N. Morris for the animal husbandry and N. Teja for assistance with cell

culture. We also thank K. Sixt for comments on the manuscript. This research is supported by the Intramural Research Program of the US National Cancer Institute, US National Institutes of

Health, Center for Cancer Research. M.Y. was partially supported by the Japan Society for the Promotion of Science grant 21689053. AUTHOR INFORMATION Author notes * Motozo Yamashita Present

address: Present address: Department of Periodontology, Osaka University Graduate School of Dentistry, Yamadaoka, Suita-Osaka, Japan., AUTHORS AND AFFILIATIONS * Laboratory of Cellular and

Molecular Biology, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland, USA Michael Blank, Yi Tang, Motozo Yamashita & Ying E Zhang * Comparative Molecular

Cytogenetics Core, Mouse Cancer Genetics Program, National Cancer Institute, Frederick, Maryland, USA Sandra S Burkett * Department of Developmental Genetics, School of Basic Medical

Science, Nanjing Medical University, Nanjing, Jiangsu, China Steven Y Cheng Authors * Michael Blank View author publications You can also search for this author inPubMed Google Scholar * Yi

Tang View author publications You can also search for this author inPubMed Google Scholar * Motozo Yamashita View author publications You can also search for this author inPubMed Google

Scholar * Sandra S Burkett View author publications You can also search for this author inPubMed Google Scholar * Steven Y Cheng View author publications You can also search for this author

inPubMed Google Scholar * Ying E Zhang View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS M.Y. and Y.T. maintained mouse colonies and

generated primary MEFs and mouse dermal fibroblasts. M.Y., S.Y.C. and Y.E.Z. observed and analyzed the spontaneous tumor formation in mice. S.S.B. performed karyotyping analyses. Y.E.Z.

analyzed the microarray data. M.B. performed all other experiments described in the manuscript. M.B. and Y.E.Z. conceived of the study, analyzed the data and wrote the paper. CORRESPONDING

AUTHOR Correspondence to Ying E Zhang. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing financial interests. SUPPLEMENTARY INFORMATION SUPPLEMENTARY TEXT AND FIGURES

Supplementary Figures 1–9, Supplementary Tables 1–5 and Supplementary Methods (PDF 2152 kb) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Blank, M.,

Tang, Y., Yamashita, M. _et al._ A tumor suppressor function of Smurf2 associated with controlling chromatin landscape and genome stability through RNF20. _Nat Med_ 18, 227–234 (2012).

https://doi.org/10.1038/nm.2596 Download citation * Received: 21 March 2011 * Accepted: 07 November 2011 * Published: 08 January 2012 * Issue Date: February 2012 * DOI:

https://doi.org/10.1038/nm.2596 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this content: Get shareable link Sorry, a shareable link is not currently

available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative