A tumor suppressor function of Smurf2 associated with controlling chromatin landscape and genome stability through RNF20

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, Smurf2 has a tumor suppression function that normally maintains genomic stability by controlling the epigenetic landscape of histone modifications through RNF20.

[1]  T. Hughes,et al.  BUR Kinase Selectively Regulates H3 K4 Trimethylation and H2B Ubiquitylation through Recruitment of the PAF Elongation Complex , 2005, Current Biology.

[2]  K. Zaret,et al.  Micrococcal Nuclease Analysis of Chromatin Structure , 2005, Current protocols in molecular biology.

[3]  J. Haveman,et al.  Clonogenic assay of cells in vitro , 2006, Nature Protocols.

[4]  福永 絵里奈.  Smurf2 induces ubiquitin-dependent degradation of Smurf1 to prevent migration of breast cancer cells , 2010 .

[5]  Eran Segal,et al.  Monoubiquitinated H2B is associated with the transcribed region of highly expressed genes in human cells , 2008, Nature Cell Biology.

[6]  Zu-Wen Sun,et al.  Ubiquitination of histone H2B regulates H3 methylation and gene silencing in yeast , 2002, Nature.

[7]  Stanley N Cohen,et al.  Smurf2 up-regulation activates telomere-dependent senescence. , 2004, Genes & development.

[8]  R. Roeder,et al.  The human homolog of yeast BRE1 functions as a transcriptional coactivator through direct activator interactions. , 2005, Molecular cell.

[9]  H. Green,et al.  QUANTITATIVE STUDIES OF THE GROWTH OF MOUSE EMBRYO CELLS IN CULTURE AND THEIR DEVELOPMENT INTO ESTABLISHED LINES , 1963, The Journal of cell biology.

[10]  Alessandra Montecucco,et al.  Cellular response to etoposide treatment. , 2007, Cancer letters.

[11]  Brian D. Strahl,et al.  A Novel Domain in Set2 Mediates RNA Polymerase II Interaction and Couples Histone H3 K36 Methylation with Transcript Elongation , 2005, Molecular and Cellular Biology.

[12]  J. Wrana,et al.  Smad7 binds to Smurf2 to form an E3 ubiquitin ligase that targets the TGF beta receptor for degradation. , 2000, Molecular cell.

[13]  Yves Pommier,et al.  γH2AX and cancer , 2008, Nature Reviews Cancer.

[14]  Y. Shiloh,et al.  Jcb: Article Introduction , 2002 .

[15]  R. Derynck,et al.  Regulation of Smad degradation and activity by Smurf2, an E3 ubiquitin ligase. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[16]  A. Püschel,et al.  Retracted: Ubiquitination of the GTPase Rap1B by the ubiquitin ligase Smurf2 is required for the establishment of neuronal polarity , 2007, The EMBO journal.

[17]  Xia Lin,et al.  Smurf2 Is a Ubiquitin E3 Ligase Mediating Proteasome-dependent Degradation of Smad2 in Transforming Growth Factor-β Signaling* 210 , 2000, The Journal of Biological Chemistry.

[18]  B. Hogan,et al.  Manipulating the mouse embryo: A laboratory manual , 1986 .

[19]  Vikki M. Weake,et al.  Histone ubiquitination: triggering gene activity. , 2008, Molecular cell.

[20]  H. Kuwano,et al.  High-level expression of the Smad ubiquitin ligase Smurf2 correlates with poor prognosis in patients with esophageal squamous cell carcinoma. , 2002, Cancer research.

[21]  L. Attisano,et al.  Regulation of the TGFβ signalling pathway by ubiquitin-mediated degradation , 2004, Oncogene.

[22]  A. Seth,et al.  An RNF11: Smurf2 complex mediates ubiquitination of the AMSH protein , 2004, Oncogene.

[23]  Laurence Florens,et al.  Histone Crosstalk between H2B Monoubiquitination and H3 Methylation Mediated by COMPASS , 2007, Cell.

[24]  J. Wrana,et al.  Regulation of Planar Cell Polarity by Smurf Ubiquitin Ligases , 2009, Cell.

[25]  A. Simm,et al.  Determination of copy number of c-Myc protein per cell by quantitative Western blotting. , 1999, Analytical biochemistry.

[26]  S. Jackson,et al.  Mammalian SUMO E3-ligases PIAS1 and PIAS4 promote responses to DNA double-strand breaks , 2009, Nature.

[27]  C. Deng,et al.  Ablation of Smurf2 reveals an inhibition in TGF‐β signalling through multiple mono‐ubiquitination of Smad3 , 2011, The EMBO journal.

[28]  Sarah E. Johnstone,et al.  Stress and the epigenetic landscape: a link to the pathobiology of human diseases? , 2010, Nature Reviews Genetics.

[29]  E. Lundberg,et al.  A Genecentric Human Protein Atlas for Expression Profiles Based on Antibodies* , 2008, Molecular & Cellular Proteomics.

[30]  Bernhard Kuster,et al.  Requirement of ATM-dependent monoubiquitylation of histone H2B for timely repair of DNA double-strand breaks. , 2011, Molecular cell.

[31]  Seung-Min Yang,et al.  Global chromatin compaction limits the strength of the DNA damage response , 2007, The Journal of cell biology.

[32]  M. Anver,et al.  Smad ubiquitination regulatory factor 2 promotes metastasis of breast cancer cells by enhancing migration and invasiveness. , 2009, Cancer research.

[33]  S. Kozubek,et al.  Chromatin structure influences the sensitivity of DNA to gamma-radiation. , 2008, Biochimica et biophysica acta.

[34]  Ali Shilatifard,et al.  Chromatin modifications by methylation and ubiquitination: implications in the regulation of gene expression. , 2006, Annual review of biochemistry.

[35]  A. Shilatifard,et al.  RAD6-Mediated Transcription-Coupled H2B Ubiquitylation Directly Stimulates H3K4 Methylation in Human Cells , 2009, Cell.

[36]  H. Tauchi,et al.  Regulation of homologous recombination by RNF20-dependent H2B ubiquitination. , 2011, Molecular cell.

[37]  C. Deng,et al.  Ubiquitin Ligase Smurf1 Controls Osteoblast Activity and Bone Homeostasis by Targeting MEKK2 for Degradation , 2005, Cell.

[38]  D. Raleigh,et al.  Histone H2B ubiquitylation disrupts local and higher order chromatin compaction , 2010, Nature chemical biology.

[39]  A. Moustakas,et al.  Regulating the stability of TGFβ receptors and Smads , 2009, Cell Research.

[40]  M. Ashburner.  A Laboratory manual , 1989 .

[41]  Y. Shiloh,et al.  Chromatin relaxation in response to DNA double-strand breaks is modulated by a novel ATM- and KAP-1 dependent pathway , 2006, Nature Cell Biology.

[42]  Paul Tempst,et al.  Monoubiquitination of human histone H2B: the factors involved and their roles in HOX gene regulation. , 2005, Molecular cell.

[43]  C. Sherr,et al.  Tumor surveillance via the ARF-p53 pathway. , 1998, Genes & development.

[44]  A. Levine,et al.  p53 alteration is a common event in the spontaneous immortalization of primary BALB/c murine embryo fibroblasts. , 1991, Genes & development.

[45]  D. Haber,et al.  Tumour-suppressor genes: evolving definitions in the genomic age , 1997, Nature Genetics.

[46]  C. Allis,et al.  Extraction, purification and analysis of histones , 2007, Nature Protocols.