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Functional Characterization of the HuR:CD83 mRNA Interaction

By Dorothea Pieper, Susann Schirmer, Alexander T. Prechtel, Ralph H. Kehlenbach, Joachim Hauber and Jan Chemnitz


Maturation of dendritic cells (DC) is characterized by expression of CD83, a surface protein that appears to be necessary for the effective activation of naïve T-cells and T-helper cells by DC. Lately it was shown that CD83 expression is regulated on the posttranscriptional level by interaction of the shuttle protein HuR with a novel posttranscriptional regulatory RNA element (PRE), which is located in the coding region of the CD83 transcript. Interestingly, this interaction commits the CD83 mRNA to efficient nuclear export via the CRM1 pathway. To date, however, the structural basis of this interaction, which potentially involves three distinct RNA recognition motifs (RRM1–3) in HuR and a complex three-pronged RNA stem-loop element in CD83 mRNA, has not been investigated in detail. In the present work we analyzed this interaction in vitro and in vivo using various HuR- and CD83 mRNA mutants. We are able to demonstrate that both, RRM1 and RRM2 are crucial for binding, whereas RRM3 as well as the HuR hinge region contributed only marginally to this protein∶RNA interaction. Furthermore, mutation of uridine rich patches within the PRE did not disturb HuR:CD83 mRNA complex formation while, in contrast, the deletion of specific PRE subfragments from the CD83 mRNA prevented HuR binding in vitro and in vivo. Interestingly, the observed inhibition of HuR binding to CD83 mRNA does not lead to a nuclear trapping of the transcript but rather redirected this transcript from the CRM1- towards the NXF1/TAP-specific nuclear export pathway. Thus, the presence of a functional PRE permits nucleocytoplasmic trafficking of the CD83 transcript via the CRM1 pathway

Topics: Research Article
Publisher: Public Library of Science
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Provided by: PubMed Central

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  1. (1995). A conserved family of elav-like genes in vertebrates.
  2. (1992). A novel cell-surface molecule expressed by human interdigitating reticulum cells, Langerhans cells, and activated lymphocytes is a new member of the Ig superfamily.
  3. (2001). A novel mRNA-decapping activity in HeLa cytoplasmic extracts is regulated by AU-rich elements.
  4. (1994). A small element from the Mason-Pfizer monkey virus genome makes human immunodeficiency virus type 1 expression and replication Rev-independent.
  5. (2005). Adenovirus E4orf6 targets pp32/LANP to control the fate of ARE-containing mRNAs by perturbing the CRM1-dependent mechanism.
  6. (2007). Analysis of nucleocytoplasmic trafficking of the HuR ligand APRIL and its influence on CD83 expression.
  7. (2002). Antigen presentation and T cell stimulation by dendritic cells.
  8. (2006). ARED 3.0: the large and diverse AU-rich transcriptome.
  9. (2005). AU-rich elements and associated factors: are there unifying principles?
  10. (1993). Biochemical characterization of binding of multiple HIV-1 Rev monomeric proteins to the Rev responsive element.
  11. (1996). CD14+ blood monocytes can differentiate into functionally mature CD83+ dendritic cells.
  12. (2008). CD83 regulates lymphocyte maturation, activation and homeostasis.
  13. (2006). CD83: a regulatory molecule of the immune system with great potential for therapeutic application.
  14. (1996). Cloning and characterization of HuR, a ubiquitously expressed Elav-like protein.
  15. (2002). Cloning and characterization of the promoter region of the human CD83 gene.
  16. (1994). Conserved structures and diversity of functions of RNA-binding proteins.
  17. (1997). CRM1 is an export receptor for leucine-rich nuclear export signals.
  18. (2004). CRM1-dependent, but not ARE-mediated, nuclear export of IFN-alpha1 mRNA.
  19. (2007). CRM1-mediated nuclear export: to the pore and beyond. Trends Cell Biol 17: 193–201.
  20. (2008). Dendritic cell CD83: a therapeutic target or innocent bystander?
  21. (1998). Dendritic cells and the control of immunity.
  22. (2001). Dendritic cells: specialized and regulated antigen processing machines.
  23. (2011). Different modes of interaction by TIAR and HuR with target RNA and DNA.
  24. (2006). Differential IFN-gamma stimulation of HLA-A gene expression through CRM-1-dependent nuclear RNA export.
  25. (1999). Direct observation of nucleocytoplasmic transport by microinjection of GFP-tagged proteins in living cells.
  26. (2008). Diverse molecular functions of Hu proteins.
  27. (2010). Dual action of epidermal growth factor: extracellular signal-stimulated nuclear-cytoplasmic export and coordinated translation of selected messenger RNA.
  28. (2006). eIF4E is a central node of an RNA regulon that governs cellular proliferation.
  29. (1998). ELAV protein HuA (HuR) can redistribute between nucleus and cytoplasm and is upregulated during serum stimulation and T cell activation.
  30. (1999). ELAV proteins stabilize deadenylated intermediates in a novel in vitro mRNA deadenylation/ degradation system.
  31. (2007). Exporting RNA from the nucleus to the cytoplasm.
  32. (1990). HIV-1 structural gene expression requires binding of the Rev trans-activator to its RNA target sequence.
  33. (1998). HNS, a nuclear-cytoplasmic shuttling sequence in HuR.
  34. (2000). HuD RNA recognition motifs play distinct roles in the formation of a stable complex with AU-rich RNA.
  35. (1991). HuD, a paraneoplastic encephalomyelitis antigen, contains RNA-binding domains and is homologous to Elav and Sex-lethal.
  36. (1995). Human blood dendritic cells selectively express CD83, a member of the immunoglobulin superfamily.
  37. (2001). HuR and mRNA stability.
  38. (2000). Immunobiology of dendritic cells.
  39. (2001). Importin-beta-like nuclear transport receptors.
  40. (2000). Inhibition of CD83 cell surface expression during dendritic cell maturation by interference with nuclear export of CD83 mRNA.
  41. Kra ¨usslich HG (2000) Multiple copies of the MasonPfizer monkey virus constitutive RNA transport element lead to enhanced HIV1 Gag expression in a context-dependent manner.
  42. (1999). Leptomycin B inactivates CRM1/exportin 1 by covalent modification at a cysteine residue in the central conserved region.
  43. (1998). Leptomycin B inhibition of signal-mediated nuclear export by direct binding to CRM1.
  44. (2003). Leptomycin B, an inhibitor of the nuclear export receptor CRM1, inhibits COX-2 expression.
  45. (1994). Mammalian homologs of Drosophila ELAV localized to a neuronal subset can bind in vitro to the 39 UTR of mRNA encoding the Id transcriptional repressor.
  46. (1998). Mapping of a minimal AU-rich sequence required for lipopolysaccharide-induced binding of a 55-kDa protein on tumor necrosis factor-alpha mRNA.
  47. (2009). mRNA nuclear export at a glance.
  48. (2010). Novel cis-active structures in the coding region mediate CRM1-dependent nuclear export of IFN-alpha 1 mRNA.
  49. (1998). Overexpression of HuR, a nuclear-cytoplasmic shuttling protein, increases the in vivo stability of ARE-containing mRNAs.
  50. (2009). Phosphorylation of the HuR ligand APRIL by casein kinase 2 regulates CD83 expression.
  51. (2000). Protein ligands to HuR modulate its interaction with target mRNAs in vivo.
  52. (1996). Purification and properties of HuD, a neuronal RNA-binding protein.
  53. (2010). Reduced nuclear export of HuR mRNA by HuR is linked to the loss of HuR in replicative senescence.
  54. (1998). RNA stabilization by the AU-rich element binding protein, HuR, an ELAV protein.
  55. (2007). Specific protein domains mediate cooperative assembly of HuR oligomers on AU-rich mRNA-destabilizing sequences.
  56. (2002). Steinkasserer A
  57. Steinkasserer A (2002) CD83 on dendritic cells: more than just a marker for maturation.
  58. Steinkasserer A (2007) CD83: an update on functions and prospects of the maturation marker of dendritic cells.
  59. (2006). Stimulated expression of mRNAs in activated T cells depends on a functional CRM1 nuclear export pathway.
  60. (2001). The 39 untranslated region of tumor necrosis factor alpha mRNA is a target of the mRNA-stabilizing factor HuR.
  61. (1991). The dendritic cell system and its role in immunogenicity.
  62. (1997). The Elav-like proteins bind to AU-rich elements and to the poly(A) tail of mRNA.
  63. (1998). The HIV-1 Rev protein.
  64. (1989). The HIV-1 rev trans-activator acts through a structured target sequence to activate nuclear export of unspliced viral mRNA.
  65. (2010). The karyopherin CRM1 is required for dendritic cell maturation.
  66. (2005). The RNA recognition motif, a plastic RNA-binding platform to regulate post-transcriptional gene expression.
  67. (1999). Titration of cellular export factors, but not heteromultimerization, is the molecular mechanism of transdominant HTLV-1 rex mutants.
  68. (1999). Why is Hu where? Shuttling of early-response-gene messenger RNA subsets.