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Binding of the Heterogeneous Ribonucleoprotein K (hnRNP K) to the Epstein-Barr Virus Nuclear Antigen 2 (EBNA2) Enhances Viral LMP2A Expression
The Epstein-Barr Virus (EBV) -encoded EBNA2 protein, which is essential for the in vitro transformation of B-lymphocytes, interferes with cellular processes by binding to proteins via conserved sequence motifs. Its Arginine-Glycine (RG) repeat element contains either symmetrically or asymmetrically...
Autores principales: | , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3411732/ https://www.ncbi.nlm.nih.gov/pubmed/22879910 http://dx.doi.org/10.1371/journal.pone.0042106 |
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author | Gross, Henrik Hennard, Christine Masouris, Ilias Cassel, Christian Barth, Stephanie Stober-Grässer, Ute Mamiani, Alfredo Moritz, Bodo Ostareck, Dirk Ostareck-Lederer, Antje Neuenkirchen, Nils Fischer, Utz Deng, Wen Leonhardt, Heinrich Noessner, Elfriede Kremmer, Elisabeth Grässer, Friedrich A. |
author_facet | Gross, Henrik Hennard, Christine Masouris, Ilias Cassel, Christian Barth, Stephanie Stober-Grässer, Ute Mamiani, Alfredo Moritz, Bodo Ostareck, Dirk Ostareck-Lederer, Antje Neuenkirchen, Nils Fischer, Utz Deng, Wen Leonhardt, Heinrich Noessner, Elfriede Kremmer, Elisabeth Grässer, Friedrich A. |
author_sort | Gross, Henrik |
collection | PubMed |
description | The Epstein-Barr Virus (EBV) -encoded EBNA2 protein, which is essential for the in vitro transformation of B-lymphocytes, interferes with cellular processes by binding to proteins via conserved sequence motifs. Its Arginine-Glycine (RG) repeat element contains either symmetrically or asymmetrically di-methylated arginine residues (SDMA and ADMA, respectively). EBNA2 binds via its SDMA-modified RG-repeat to the survival motor neurons protein (SMN) and via the ADMA-RG-repeat to the NP9 protein of the human endogenous retrovirus K (HERV-K (HML-2) Type 1). The hypothesis of this work was that the methylated RG-repeat mimics an epitope shared with cellular proteins that is used for interaction with target structures. With monoclonal antibodies against the modified RG-repeat, we indeed identified cellular homologues that apparently have the same surface structure as methylated EBNA2. With the SDMA-specific antibodies, we precipitated the Sm protein D3 (SmD3) which, like EBNA2, binds via its SDMA-modified RG-repeat to SMN. With the ADMA-specific antibodies, we precipitated the heterogeneous ribonucleoprotein K (hnRNP K). Specific binding of the ADMA- antibody to hnRNP K was demonstrated using E. coli expressed/ADMA-methylated hnRNP K. In addition, we show that EBNA2 and hnRNP K form a complex in EBV- infected B-cells. Finally, hnRNP K, when co-expressed with EBNA2, strongly enhances viral latent membrane protein 2A (LMP2A) expression by an unknown mechanism as we did not detect a direct association of hnRNP K with DNA-bound EBNA2 in gel shift experiments. Our data support the notion that the methylated surface of EBNA2 mimics the surface structure of cellular proteins to interfere with or co-opt their functional properties. |
format | Online Article Text |
id | pubmed-3411732 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-34117322012-08-09 Binding of the Heterogeneous Ribonucleoprotein K (hnRNP K) to the Epstein-Barr Virus Nuclear Antigen 2 (EBNA2) Enhances Viral LMP2A Expression Gross, Henrik Hennard, Christine Masouris, Ilias Cassel, Christian Barth, Stephanie Stober-Grässer, Ute Mamiani, Alfredo Moritz, Bodo Ostareck, Dirk Ostareck-Lederer, Antje Neuenkirchen, Nils Fischer, Utz Deng, Wen Leonhardt, Heinrich Noessner, Elfriede Kremmer, Elisabeth Grässer, Friedrich A. PLoS One Research Article The Epstein-Barr Virus (EBV) -encoded EBNA2 protein, which is essential for the in vitro transformation of B-lymphocytes, interferes with cellular processes by binding to proteins via conserved sequence motifs. Its Arginine-Glycine (RG) repeat element contains either symmetrically or asymmetrically di-methylated arginine residues (SDMA and ADMA, respectively). EBNA2 binds via its SDMA-modified RG-repeat to the survival motor neurons protein (SMN) and via the ADMA-RG-repeat to the NP9 protein of the human endogenous retrovirus K (HERV-K (HML-2) Type 1). The hypothesis of this work was that the methylated RG-repeat mimics an epitope shared with cellular proteins that is used for interaction with target structures. With monoclonal antibodies against the modified RG-repeat, we indeed identified cellular homologues that apparently have the same surface structure as methylated EBNA2. With the SDMA-specific antibodies, we precipitated the Sm protein D3 (SmD3) which, like EBNA2, binds via its SDMA-modified RG-repeat to SMN. With the ADMA-specific antibodies, we precipitated the heterogeneous ribonucleoprotein K (hnRNP K). Specific binding of the ADMA- antibody to hnRNP K was demonstrated using E. coli expressed/ADMA-methylated hnRNP K. In addition, we show that EBNA2 and hnRNP K form a complex in EBV- infected B-cells. Finally, hnRNP K, when co-expressed with EBNA2, strongly enhances viral latent membrane protein 2A (LMP2A) expression by an unknown mechanism as we did not detect a direct association of hnRNP K with DNA-bound EBNA2 in gel shift experiments. Our data support the notion that the methylated surface of EBNA2 mimics the surface structure of cellular proteins to interfere with or co-opt their functional properties. Public Library of Science 2012-08-03 /pmc/articles/PMC3411732/ /pubmed/22879910 http://dx.doi.org/10.1371/journal.pone.0042106 Text en © 2012 Gross et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Gross, Henrik Hennard, Christine Masouris, Ilias Cassel, Christian Barth, Stephanie Stober-Grässer, Ute Mamiani, Alfredo Moritz, Bodo Ostareck, Dirk Ostareck-Lederer, Antje Neuenkirchen, Nils Fischer, Utz Deng, Wen Leonhardt, Heinrich Noessner, Elfriede Kremmer, Elisabeth Grässer, Friedrich A. Binding of the Heterogeneous Ribonucleoprotein K (hnRNP K) to the Epstein-Barr Virus Nuclear Antigen 2 (EBNA2) Enhances Viral LMP2A Expression |
title | Binding of the Heterogeneous Ribonucleoprotein K (hnRNP K) to the Epstein-Barr Virus Nuclear Antigen 2 (EBNA2) Enhances Viral LMP2A Expression |
title_full | Binding of the Heterogeneous Ribonucleoprotein K (hnRNP K) to the Epstein-Barr Virus Nuclear Antigen 2 (EBNA2) Enhances Viral LMP2A Expression |
title_fullStr | Binding of the Heterogeneous Ribonucleoprotein K (hnRNP K) to the Epstein-Barr Virus Nuclear Antigen 2 (EBNA2) Enhances Viral LMP2A Expression |
title_full_unstemmed | Binding of the Heterogeneous Ribonucleoprotein K (hnRNP K) to the Epstein-Barr Virus Nuclear Antigen 2 (EBNA2) Enhances Viral LMP2A Expression |
title_short | Binding of the Heterogeneous Ribonucleoprotein K (hnRNP K) to the Epstein-Barr Virus Nuclear Antigen 2 (EBNA2) Enhances Viral LMP2A Expression |
title_sort | binding of the heterogeneous ribonucleoprotein k (hnrnp k) to the epstein-barr virus nuclear antigen 2 (ebna2) enhances viral lmp2a expression |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3411732/ https://www.ncbi.nlm.nih.gov/pubmed/22879910 http://dx.doi.org/10.1371/journal.pone.0042106 |
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