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PMID: 18523580 Published · epublish English Journal Article Research Support, Non-U.S. Gov't

The Polycomb protein and E3 ubiquitin ligase Ring1B harbors an IRES in its highly conserved 5' UTR.

PloS one ·Vol. 3 ·No. 6 ·2008-06-04 ·Pages e2322

Boutsma E, Noback S, van Lohuizen M

Abstract

Ring1B is an essential member of the highly conserved Polycomb group proteins, which orchestrate developmental processes, cell growth and stem cell fate by modifying local chromatin structure. Ring1B was found to be the E3 ligase that monoubiquitinates histone H2A, which adds a new level of chromatin modification to Polycomb group proteins. Here we report that Ring1B belongs to the exclusive group of proteins that for their translation depend on a stable 5' UTR sequence in their mRNA known as an Internal Ribosome Entry Site (IRES). In cell transfection assays the Ring1B IRES confers significantly higher expression levels of Ring1B than a Ring1B cDNA without the IRES. Also, dual luciferase assays show strong activity of the Ring1B IRES. Although our findings indicate Ring1B can be translated under conditions where cap-dependent translation is impaired, we found the Ring1B IRES to be cap-dependent. This raises the possibility that translational control of Ring1B is a multi-layered process and that translation of Ring1B needs to be maintained under varying conditions, which is in line with its essential role as an E3 ligase for monoubiquitination of histone H2A in the PRC1 Polycomb protein complex.

MeSH Terms
5' Untranslated Regions Animals Base Sequence Blotting, Western Cell Line DNA Primers Expressed Sequence Tags Point Mutation Polycomb Repressive Complex 1 Polycomb-Group Proteins Polymerase Chain Reaction Promoter Regions, Genetic RNA, Messenger/genetics Repressor Proteins/chemistry,genetics Ribosomes Ubiquitin-Protein Ligases/chemistry,genetics
Chemicals
5' Untranslated Regions DNA Primers Polycomb-Group Proteins RNA, Messenger Repressor Proteins Polycomb Repressive Complex 1 Ubiquitin-Protein Ligases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Boutsma Erwin
Division of Molecular Genetics, Netherlands Cancer Institute, Amsterdam, The Netherlands.
Noback Sonja
van Lohuizen Maarten
References (63)
63 references, click to expand
  1. Genome-wide analysis of Polycomb targets in Drosophila melanogaster.
    Nat Genet. 2006 Jun;38(6):700-5 PMID: 16732288
  2. Control of developmental regulators by Polycomb in human embryonic stem cells.
    Cell. 2006 Apr 21;125(2):301-13 PMID: 16630818
  3. SETting the stage. Eed-Enx1 leaves an epigenetic signature on the inactive X chromosome.
    Dev Cell. 2003 Apr;4(4):445-7 PMID: 12689584
  4. MLL repression domain interacts with histone deacetylases, the polycomb group proteins HPC2 and BMI-1, and the corepressor C-terminal-binding protein.
    Proc Natl Acad Sci U S A. 2003 Jul 8;100(14):8342-7 PMID: 12829790
  5. Loss- and gain-of-function mutations show a polycomb group function for Ring1A in mice.
    Development. 2000 Dec;127(23):5093-100 PMID: 11060235
  6. Genome-wide profiling of PRC1 and PRC2 Polycomb chromatin binding in Drosophila melanogaster.
    Nat Genet. 2006 Jun;38(6):694-9 PMID: 16628213
  7. Polycomb complexes repress developmental regulators in murine embryonic stem cells.
    Nature. 2006 May 18;441(7091):349-53 PMID: 16625203
  8. Internal ribosome entry segment-mediated initiation of c-Myc protein synthesis following genotoxic stress.
    Biochem J. 2001 Oct 1;359(Pt 1):183-92 PMID: 11563982
  9. Structure and E3-ligase activity of the Ring-Ring complex of polycomb proteins Bmi1 and Ring1b.
    EMBO J. 2006 Jun 7;25(11):2465-74 PMID: 16710298
  10. Mitotically stable association of polycomb group proteins eed and enx1 with the inactive x chromosome in trophoblast stem cells.
    Curr Biol. 2002 Jun 25;12(12):1016-20 PMID: 12123576
  11. Specific roles of 5' RNA secondary structures in stabilizing transcripts in chloroplasts.
    Nucleic Acids Res. 2005 Aug 22;33(15):4754-61 PMID: 16116040
  12. Stem cells and cancer; the polycomb connection.
    Cell. 2004 Aug 20;118(4):409-18 PMID: 15315754
  13. Cleavage of eIF4G by HIV-1 protease: effects on translation.
    FEBS Lett. 2003 Jan 2;533(1-3):89-94 PMID: 12505164
  14. Phosphorylation of initiation factor-2 alpha is required for activation of internal translation initiation during cell differentiation.
    Eur J Biochem. 2002 Jun;269(11):2810-9 PMID: 12047392
  15. Polypyrimidine tract binding protein and poly r(C) binding protein 1 interact with the BAG-1 IRES and stimulate its activity in vitro and in vivo.
    Nucleic Acids Res. 2003 Jan 15;31(2):639-46 PMID: 12527772
  16. Ring1b-mediated H2A ubiquitination associates with inactive X chromosomes and is involved in initiation of X inactivation.
    J Biol Chem. 2004 Dec 17;279(51):52812-5 PMID: 15509584
  17. RNAdraw: an integrated program for RNA secondary structure calculation and analysis under 32-bit Microsoft Windows.
    Comput Appl Biosci. 1996 Jun;12(3):247-9 PMID: 8872395
  18. Polycomb group proteins Ring1A/B are functionally linked to the core transcriptional regulatory circuitry to maintain ES cell identity.
    Development. 2008 Apr;135(8):1513-24 PMID: 18339675
  19. The Polycomb group protein Eed protects the inactive X-chromosome from differentiation-induced reactivation.
    Nat Cell Biol. 2006 Feb;8(2):195-202 PMID: 16415857
  20. Heterogeneous nuclear ribonucleoprotein C modulates translation of c-myc mRNA in a cell cycle phase-dependent manner.
    Mol Cell Biol. 2003 Jan;23(2):708-20 PMID: 12509468
  21. Assessing IRES activity in the HIF-1alpha and other cellular 5' UTRs.
    RNA. 2006 Jun;12(6):1074-83 PMID: 16601206
  22. Role of histone H3 lysine 27 methylation in Polycomb-group silencing.
    Science. 2002 Nov 1;298(5595):1039-43 PMID: 12351676
  23. Stable X chromosome inactivation involves the PRC1 Polycomb complex and requires histone MACROH2A1 and the CULLIN3/SPOP ubiquitin E3 ligase.
    Proc Natl Acad Sci U S A. 2005 May 24;102(21):7635-40 PMID: 15897469
  24. Posterior transformation, neurological abnormalities, and severe hematopoietic defects in mice with a targeted deletion of the bmi-1 proto-oncogene.
    Genes Dev. 1994 Apr 1;8(7):757-69 PMID: 7926765
  25. Role of histone H2A ubiquitination in Polycomb silencing.
    Nature. 2004 Oct 14;431(7010):873-8 PMID: 15386022
  26. Transcriptional repression mediated by the human polycomb-group protein EED involves histone deacetylation.
    Nat Genet. 1999 Dec;23(4):474-8 PMID: 10581039
  27. Demonstrating internal ribosome entry sites in eukaryotic mRNAs using stringent RNA test procedures.
    RNA. 2004 Apr;10(4):720-30 PMID: 15037781
  28. Internal ribosome entry site-mediated translation of a mammalian mRNA is regulated by amino acid availability.
    J Biol Chem. 2001 Apr 13;276(15):12285-91 PMID: 11114306
  29. rgr oncogene: activation by elimination of translational controls and mislocalization.
    Cancer Res. 2003 Jul 15;63(14):4188-95 PMID: 12874025
  30. Recruitment of PRC1 function at the initiation of X inactivation independent of PRC2 and silencing.
    EMBO J. 2006 Jul 12;25(13):3110-22 PMID: 16763550
  31. Translational enhancement of the poliovirus 5' noncoding region mediated by virus-encoded polypeptide 2A.
    Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10272-6 PMID: 1332040
  32. Genome-wide mapping of Polycomb target genes unravels their roles in cell fate transitions.
    Genes Dev. 2006 May 1;20(9):1123-36 PMID: 16618801
  33. Characterization of interactions between the mammalian polycomb-group proteins Enx1/EZH2 and EED suggests the existence of different mammalian polycomb-group protein complexes.
    Mol Cell Biol. 1998 Jun;18(6):3586-95 PMID: 9584199
  34. Histone methyltransferase activity associated with a human multiprotein complex containing the Enhancer of Zeste protein.
    Genes Dev. 2002 Nov 15;16(22):2893-905 PMID: 12435631
  35. Hypoxia-inducible factor-1alpha mRNA contains an internal ribosome entry site that allows efficient translation during normoxia and hypoxia.
    Mol Biol Cell. 2002 May;13(5):1792-801 PMID: 12006670
  36. IRES interaction with translation initiation factors: functional characterization of novel RNA contacts with eIF3, eIF4B, and eIF4GII.
    RNA. 2001 Sep;7(9):1213-26 PMID: 11565745
  37. Disappearance of ubiquitinated histone H2A during chromatin condensation in TGF beta 1-induced apoptosis.
    Anticancer Res. 1995 Mar-Apr;15(2):267-72 PMID: 7762993
  38. Bmi-1 determines the proliferative capacity of normal and leukaemic stem cells.
    Nature. 2003 May 15;423(6937):255-60 PMID: 12714970
  39. Translation of vascular endothelial growth factor mRNA by internal ribosome entry: implications for translation under hypoxia.
    Mol Cell Biol. 1998 Jun;18(6):3112-9 PMID: 9584152
  40. Caspase-dependent deubiquitination of monoubiquitinated nucleosomal histone H2A induced by diverse apoptogenic stimuli.
    Cell Death Differ. 2001 Dec;8(12):1182-96 PMID: 11753566
  41. Rnf2 (Ring1b) deficiency causes gastrulation arrest and cell cycle inhibition.
    Proc Natl Acad Sci U S A. 2003 Mar 4;100(5):2468-73 PMID: 12589020
  42. The Polycomb group protein EED is dispensable for the initiation of random X-chromosome inactivation.
    PLoS Genet. 2006 May;2(5):e66 PMID: 16680199
  43. Heterogeneous nuclear ribonucleoprotein A1 is a novel internal ribosome entry site trans-acting factor that modulates alternative initiation of translation of the fibroblast growth factor 2 mRNA.
    J Biol Chem. 2005 Feb 11;280(6):4144-53 PMID: 15525641
  44. Bmi1 is essential for cerebellar development and is overexpressed in human medulloblastomas.
    Nature. 2004 Mar 18;428(6980):337-41 PMID: 15029199
  45. The functions of E(Z)/EZH2-mediated methylation of lysine 27 in histone H3.
    Curr Opin Genet Dev. 2004 Apr;14(2):155-64 PMID: 15196462
  46. Structure of a Bmi-1-Ring1B polycomb group ubiquitin ligase complex.
    J Biol Chem. 2006 Jul 21;281(29):20643-9 PMID: 16714294
  47. Role of Bmi-1 and Ring1A in H2A ubiquitylation and Hox gene silencing.
    Mol Cell. 2005 Dec 22;20(6):845-54 PMID: 16359901
  48. Poliovirus 2A protease induces apoptotic cell death.
    Mol Cell Biol. 2000 Feb;20(4):1271-7 PMID: 10648613
  49. The core of the polycomb repressive complex is compositionally and functionally conserved in flies and humans.
    Mol Cell Biol. 2002 Sep;22(17):6070-8 PMID: 12167701
  50. A role for mel-18, a Polycomb group-related vertebrate gene, during theanteroposterior specification of the axial skeleton.
    Development. 1996 May;122(5):1513-22 PMID: 8625838
  51. Enhanced self-renewal of hematopoietic stem cells mediated by the polycomb gene product Bmi-1.
    Immunity. 2004 Dec;21(6):843-51 PMID: 15589172
  52. The E2F6 transcription factor is a component of the mammalian Bmi1-containing polycomb complex.
    Proc Natl Acad Sci U S A. 2001 Feb 13;98(4):1519-24 PMID: 11171983
  53. Targeted disruption of the mouse homologue of the Drosophila polyhomeotic gene leads to altered anteroposterior patterning and neural crest defects.
    Development. 1997 Oct;124(19):3673-82 PMID: 9367423
  54. Translation of eukaryotic translation initiation factor 4GI (eIF4GI) proceeds from multiple mRNAs containing a novel cap-dependent internal ribosome entry site (IRES) that is active during poliovirus infection.
    J Biol Chem. 2005 May 13;280(19):18610-22 PMID: 15755734
  55. The Drosophila Polycomb Group proteins ESC and E(Z) are present in a complex containing the histone-binding protein p55 and the histone deacetylase RPD3.
    Development. 2001 Jan;128(2):275-86 PMID: 11124122
  56. Bmi-1 dependence distinguishes neural stem cell self-renewal from progenitor proliferation.
    Nature. 2003 Oct 30;425(6961):962-7 PMID: 14574365
  57. Internet resources for the functional analysis of 5' and 3' untranslated regions of eukaryotic mRNAs.
    Trends Genet. 1999 Sep;15(9):378 PMID: 10461207
  58. Altered cellular proliferation and mesoderm patterning in Polycomb-M33-deficient mice.
    Development. 1997 Feb;124(3):721-9 PMID: 9043087
  59. Interaction of mouse polycomb-group (Pc-G) proteins Enx1 and Enx2 with Eed: indication for separate Pc-G complexes.
    Mol Cell Biol. 1998 Jun;18(6):3572-9 PMID: 9584197
  60. Chromatin-association of the Polycomb group protein BMI1 is cell cycle-regulated and correlates with its phosphorylation status.
    J Cell Sci. 1999 Dec;112 ( Pt 24):4627-39 PMID: 10574711
  61. Ring1-mediated ubiquitination of H2A restrains poised RNA polymerase II at bivalent genes in mouse ES cells.
    Nat Cell Biol. 2007 Dec;9(12):1428-35 PMID: 18037880
  62. Establishment of histone h3 methylation on the inactive X chromosome requires transient recruitment of Eed-Enx1 polycomb group complexes.
    Dev Cell. 2003 Apr;4(4):481-95 PMID: 12689588
  63. c-Myc protein synthesis is initiated from the internal ribosome entry segment during apoptosis.
    Mol Cell Biol. 2000 Feb;20(4):1162-9 PMID: 10648601
Article Info
Journal
PloS one
Abbr.
PLoS One
ISSN
1932-6203
DOI
10.1371/journal.pone.0002322
Published
2008-06-04
Epub
2008-00-04
Pages
e2322
Language
English
Region
United States
NLM ID
101285081
PMCID
PMC2386971
Subset
IM
External Links
PubMed source DOI: 10.1371/journal.pone.0002322 PMC Full Text View journal details
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