Monoubiquitination of human histone H2B: the factors involved and their roles in HOX gene regulation.

In yeast, histone H2B monoubiquitination is a cotranscriptional event regulating histone H3 methylation at lysines 4 and 79. However, mammalian H2B monoubiquitination remains poorly understood. We report that in humans, the 600 kDa RNF20/40 complex is the E3 ligase and UbcH6 is the ubiquitin E2-conjugating enzyme for H2B-Lys120 monoubiquitination. RNF20 and RNF40 are both homologs of Bre1, the E3 ligase in the yeast case. UbcH6 physically interacts with RNF20/40 and with the hPAF complex. Formation of a trimeric complex with hPAF stimulates H2B monoubiquitination activity in vitro. Accordingly, UbcH6, RNF20/40, and the hPAF complex are recruited to transcriptionally active genes in vivo. RNF20 overexpression leads to elevated H2B monoubiquitination, subsequently higher levels of methylation at H3 lysines 4 and 79, and stimulation of HOX gene expression. In contrast, RNAi against the RNF20/40 complex or hPAF complex reduces H2B monoubiquitination, lowers methylation levels at H3 lysines 4 and 79, and represses HOX gene expression.

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

[2]  M. Oren,et al.  The RING domain of Mdm2 mediates histone ubiquitylation and transcriptional repression. , 2004, Molecular cell.

[3]  P. Sautière,et al.  The structure of ubiquitinated histone H2B. , 1987, The EMBO journal.

[4]  Kevin Struhl,et al.  The Rtf1 Component of the Paf1 Transcriptional Elongation Complex Is Required for Ubiquitination of Histone H2B* , 2003, Journal of Biological Chemistry.

[5]  G. Orphanides,et al.  A Unified Theory of Gene Expression , 2002, Cell.

[6]  O. de Backer,et al.  The activation of human gene MAGE-1 in tumor cells is correlated with genome-wide demethylation. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[7]  G. Kay,et al.  Menin associates with a trithorax family histone methyltransferase complex and with the hoxc8 locus. , 2004, Molecular cell.

[8]  M. Johnston,et al.  The Paf1 complex is required for histone H3 methylation by COMPASS and Dot1p: linking transcriptional elongation to histone methylation. , 2003, Molecular cell.

[9]  S. Korsmeyer,et al.  Altered Hox expression and segmental identity in Mll-mutant mice , 1995, Nature.

[10]  W. Herr,et al.  Human Sin3 deacetylase and trithorax-related Set1/Ash2 histone H3-K4 methyltransferase are tethered together selectively by the cell-proliferation factor HCF-1. , 2003, Genes & development.

[11]  C. Allis,et al.  Histone and chromatin cross-talk. , 2003, Current opinion in cell biology.

[12]  Mark Gerber,et al.  Transcriptional Elongation by RNA Polymerase II and Histone Methylation* , 2003, Journal of Biological Chemistry.

[13]  Yi Zhang,et al.  Bre1, an E3 ubiquitin ligase required for recruitment and substrate selection of Rad6 at a promoter. , 2003, Molecular cell.

[14]  Thomas A Milne,et al.  MLL targets SET domain methyltransferase activity to Hox gene promoters. , 2002, Molecular cell.

[15]  R. Amasino,et al.  PAF1-complex-mediated histone methylation of FLOWERING LOCUS C chromatin is required for the vernalization-responsive, winter-annual habit in Arabidopsis. , 2004, Genes & development.

[16]  Charles Boone,et al.  A conserved RING finger protein required for histone H2B monoubiquitination and cell size control. , 2003, Molecular cell.

[17]  M. Kashlev,et al.  Nucleosome remodeling induced by RNA polymerase II: loss of the H2A/H2B dimer during transcription. , 2002, Molecular cell.

[18]  O. Rozenblatt-Rosen,et al.  The Parafibromin Tumor Suppressor Protein Is Part of a Human Paf1 Complex , 2005, Molecular and Cellular Biology.

[19]  R. Kingston,et al.  Cooperation between Complexes that Regulate Chromatin Structure and Transcription , 2002, Cell.

[20]  Paul Tempst,et al.  The human PAF complex coordinates transcription with events downstream of RNA synthesis. , 2005, Genes & development.

[21]  Ali Shilatifard,et al.  Transcriptional activation via sequential histone H2B ubiquitylation and deubiquitylation, mediated by SAGA-associated Ubp8. , 2003, Genes & development.

[22]  G. Orphanides,et al.  FACT Facilitates Transcription-Dependent Nucleosome Alteration , 2003, Science.

[23]  R. Norum,et al.  Hereditary hyperparathyroidism and multiple ossifying jaw fibromas: a clinically and genetically distinct syndrome. , 1990, Surgery.

[24]  Michael Hampsey,et al.  Tails of Intrigue Phosphorylation of RNA Polymerase II Mediates Histone Methylation , 2003, Cell.

[25]  Thomas A. Milne,et al.  Physical Association and Coordinate Function of the H3 K4 Methyltransferase MLL1 and the H4 K16 Acetyltransferase MOF , 2005, Cell.

[26]  Mark Johnston,et al.  The Paf1 Complex Is Essential for Histone Monoubiquitination by the Rad6-Bre1 Complex, Which Signals for Histone Methylation by COMPASS and Dot1p* , 2003, Journal of Biological Chemistry.

[27]  D. Reinberg,et al.  Transcription regulation by histone methylation: interplay between different covalent modifications of the core histone tails. , 2001, Genes & development.

[28]  J. Carpten,et al.  Somatic and germ-line mutations of the HRPT2 gene in sporadic parathyroid carcinoma. , 2003, The New England journal of medicine.

[29]  Danny Reinberg,et al.  Elongation by RNA polymerase II: the short and long of it. , 2004, Genes & development.

[30]  M. Gstaiger,et al.  The HRPT2 Tumor Suppressor Gene Product Parafibromin Associates with Human PAF1 and RNA Polymerase II , 2005, Molecular and Cellular Biology.

[31]  C. Allis,et al.  Translating the Histone Code , 2001, Science.

[32]  Michael Grunstein,et al.  Histone acetylation and deacetylation in yeast , 2003, Nature Reviews Molecular Cell Biology.

[33]  A. Levine,et al.  The mdm-2 oncogene product forms a complex with the p53 protein and inhibits p53-mediated transactivation , 1992, Cell.

[34]  M. Greaves,et al.  In utero rearrangements in the trithorax-related oncogene in infant leukaemias , 1993, Nature.

[35]  Tony Kouzarides,et al.  Histone H3 lysine 4 methylation patterns in higher eukaryotic genes , 2004, Nature Cell Biology.

[36]  Richard A Young,et al.  Exchange of RNA polymerase II initiation and elongation factors during gene expression in vivo. , 2002, Molecular cell.

[37]  W. Herr,et al.  Leukemia Proto-Oncoprotein MLL Forms a SET1-Like Histone Methyltransferase Complex with Menin To Regulate Hox Gene Expression , 2004, Molecular and Cellular Biology.

[38]  Stuart L. Schreiber,et al.  Active genes are tri-methylated at K4 of histone H3 , 2002, Nature.

[39]  N. Krogan,et al.  Histone H2B Ubiquitylation Is Associated with Elongating RNA Polymerase II , 2005, Molecular and Cellular Biology.

[40]  J. Jaehning,et al.  The Yeast Paf1-RNA Polymerase II Complex Is Required for Full Expression of a Subset of Cell Cycle-Regulated Genes , 2002, Eukaryotic Cell.

[41]  Karl Henry,et al.  Rad6 plays a role in transcriptional activation through ubiquitylation of histone H2B. , 2004, Genes & development.

[42]  J. Carpten,et al.  HRPT2, encoding parafibromin, is mutated in hyperparathyroidism–jaw tumor syndrome , 2002, Nature Genetics.

[43]  Kevin Struhl,et al.  Evidence for Eviction and Rapid Deposition of Histones upon Transcriptional Elongation by RNA Polymerase II , 2004, Molecular and Cellular Biology.

[44]  Yi Zhang,et al.  hDOT1L Links Histone Methylation to Leukemogenesis , 2005, Cell.

[45]  D. Reinberg,et al.  The key to development: interpreting the histone code? , 2005, Current opinion in genetics & development.

[46]  K Hammje,et al.  HRPT2 mutations are associated with malignancy in sporadic parathyroid tumours , 2003, Journal of medical genetics.

[47]  S. Carr,et al.  Examination of micro-tip reversed-phase liquid chromatographic extraction of peptide pools for mass spectrometric analysis. , 1998, Journal of chromatography. A.

[48]  M. Osley.  H2B ubiquitylation: the end is in sight. , 2004, Biochimica et biophysica acta.

[49]  Richard A Young,et al.  Global and Hox-specific roles for the MLL1 methyltransferase. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[50]  J. Rowley,et al.  Identification of a gene, MLL, that spans the breakpoint in 11q23 translocations associated with human leukemias. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[51]  H. Erdjument-Bromage,et al.  Isolation and mass spectrometry of transcription factor complexes. , 2002, Methods.

[52]  M. Vidal,et al.  Role of histone H2A ubiquitination in Polycomb silencing , 2004, Nature.

[53]  S. van Nocker,et al.  A Mechanism Related to the Yeast Transcriptional Regulator Paf1c Is Required for Expression of the Arabidopsis FLC/MAF MADS Box Gene Familyw⃞ , 2004, The Plant Cell Online.

[54]  M. Osley,et al.  Rad6-dependent ubiquitination of histone H2B in yeast. , 2000, Science.

[55]  Craig D. Kaplan,et al.  Transcription Elongation Factors Repress Transcription Initiation from Cryptic Sites , 2003, Science.

[56]  Thomas A. Milne,et al.  WDR5 Associates with Histone H3 Methylated at K4 and Is Essential for H3 K4 Methylation and Vertebrate Development , 2005, Cell.