Structure and functional analysis of the RNA- and viral phosphoprotein-binding domain of respiratory syncytial virus M2-1 protein.
| Title: | Structure and functional analysis of the RNA- and viral phosphoprotein-binding domain of respiratory syncytial virus M2-1 protein. |
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| Authors: | Marie-Lise Blondot, Virginie Dubosclard, Jenna Fix, Safa Lassoued, Magali Aumont-Nicaise, François Bontems, Jean-François Eléouët, Christina Sizun |
| Source: | PLoS Pathogens, Vol 8, Iss 5, p e1002734 (2012) |
| Publisher Information: | Public Library of Science (PLoS), 2012. |
| Publication Year: | 2012 |
| Collection: | LCC:Immunologic diseases. Allergy LCC:Biology (General) |
| Subject Terms: | Immunologic diseases. Allergy, RC581-607, Biology (General), QH301-705.5 |
| More Details: | Respiratory syncytial virus (RSV) protein M2-1 functions as an essential transcriptional cofactor of the viral RNA-dependent RNA polymerase (RdRp) complex by increasing polymerase processivity. M2-1 is a modular RNA binding protein that also interacts with the viral phosphoprotein P, another component of the RdRp complex. These binding properties are related to the core region of M2-1 encompassing residues S58 to K177. Here we report the NMR structure of the RSV M2-1(58-177) core domain, which is structurally homologous to the C-terminal domain of Ebola virus VP30, a transcription co-factor sharing functional similarity with M2-1. The partial overlap of RNA and P interaction surfaces on M2-1(58-177), as determined by NMR, rationalizes the previously observed competitive behavior of RNA versus P. Using site-directed mutagenesis, we identified eight residues located on these surfaces that are critical for an efficient transcription activity of the RdRp complex. Single mutations of these residues disrupted specifically either P or RNA binding to M2-1 in vitro. M2-1 recruitment to cytoplasmic inclusion bodies, which are regarded as sites of viral RNA synthesis, was impaired by mutations affecting only binding to P, but not to RNA, suggesting that M2-1 is associated to the holonucleocapsid by interacting with P. These results reveal that RNA and P binding to M2-1 can be uncoupled and that both are critical for the transcriptional antitermination function of M2-1. |
| Document Type: | article |
| File Description: | electronic resource |
| Language: | English |
| ISSN: | 1553-7366 1553-7374 |
| Relation: | http://europepmc.org/articles/PMC3364950?pdf=render; https://doaj.org/toc/1553-7366; https://doaj.org/toc/1553-7374 |
| DOI: | 10.1371/journal.ppat.1002734 |
| Access URL: | https://doaj.org/article/9ce5ff1c455d4da78dc075aead552d90 |
| Accession Number: | edsdoj.9ce5ff1c455d4da78dc075aead552d90 |
| Database: | Directory of Open Access Journals |
| ISSN: | 15537366 15537374 |
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| DOI: | 10.1371/journal.ppat.1002734 |
| Published in: | PLoS Pathogens |
| Language: | English |