The matrix protein of Human respiratory syncytial virus localises to the nucleus of infected cells and inhibits transcription

Human respiratory syncytial virus (HRSV) is an enveloped RNA virus that assembles and buds from the plasma membrane of infected cells. The ribonucleoprotein complex (RNP) must associate with the viral matrix protein and glycoproteins to form newly infectious particles prior to budding. The viral proteins involved in HRSV assembly and egress are mostly unexplored. We investigated whether the glycoproteins of HRSV were involved in the late stages of viral replication by utilizing recombinant viruses where each individual glycoprotein gene was deleted and replaced with a reporter gene to maintain wild-type levels of gene expression. These engineered viruses allowed us to study the roles of the glycoproteins in assembly and budding in the context of infectious virus. Microscopy data showed that the F glycoprotein was involved in the localization of the glycoproteins with the other viral proteins at the plasma membrane. Biochemical analyses showed that deletion of the F and G proteins affected incorporation of the other viral proteins into budded virions. However, efficient viral release was unaffected by the deletion of any of the glycoproteins individually or in concert. These studies attribute a novel role to the F and G proteins in viral protein localization and assembly.

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Respiratory syncytial virus matrix protein associates with nucleocapsids in infected cells

Journal of General Virology ◽

10.1099/0022-1317-83-4-753 ◽

2002 ◽

Vol 83 (4) ◽

pp. 753-757 ◽

Cited By ~ 72

Author(s):

R. Ghildyal ◽

J. Mills ◽

M. Murray ◽

N. Vardaxis ◽

J. Meanger

Keyword(s):

Respiratory Syncytial Virus ◽

Matrix Protein ◽

Rna Viruses ◽

M Protein ◽

Dependent Manner ◽

Cytoplasmic Inclusions ◽

Negative Strand ◽

The Matrix ◽

Infected Cells ◽

Syncytial Virus

Little is known about the functions of the matrix (M) protein of respiratory syncytial virus (RSV). By analogy with other negative-strand RNA viruses, the M protein should inhibit the viral polymerase prior to packaging and facilitate virion assembly. In this study, localization of the RSV M protein in infected cells and its association with the RSV nucleocapsid complex was investigated. RSV-infected cells were shown to contain characteristic cytoplasmic inclusions. Further analysis showed that these inclusions were localization sites of the M protein as well as the N, P, L and M2-1 proteins described previously. The M protein co-purified with viral ribonucleoproteins (RNPs) from RSV-infected cells. The transcriptase activity of purified RNPs was enhanced by treatment with antibodies to the M protein in a dose-dependent manner. These data suggest that the M protein is associated with RSV nucleocapsids and, like the matrix proteins of other negative-strand RNA viruses, can inhibit virus transcription.

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RAGE inhibits human respiratory syncytial virus syncytium formation by interfering with F-protein function

Journal of General Virology ◽

10.1099/vir.0.049254-0 ◽

2013 ◽

Vol 94 (8) ◽

pp. 1691-1700 ◽

Cited By ~ 7

Author(s):

Jane Tian ◽

Kelly Huang ◽

Subramaniam Krishnan ◽

Catherine Svabek ◽

Daniel C. Rowe ◽

...

Keyword(s):

Respiratory Syncytial Virus ◽

Epithelial Cells ◽

Syncytium Formation ◽

Human Respiratory Syncytial Virus ◽

Host Cells ◽

Type I ◽

F Protein ◽

Viral Spread ◽

Infected Cells ◽

Syncytial Virus

Human respiratory syncytial virus (RSV) is a major cause of severe lower respiratory tract infection. Infection is critically dependent on the RSV fusion (F) protein, which mediates fusion between the viral envelope and airway epithelial cells. The F protein is also expressed on infected cells and is responsible for fusion of infected cells with adjacent cells, resulting in the formation of multinucleate syncytia. The receptor for advanced glycation end products (RAGE) is a pattern-recognition receptor that is constitutively highly expressed by type I alveolar epithelial cells. Here, we report that RAGE protected HEK cells from RSV-induced cell death and reduced viral titres in vitro. RAGE appeared to interact directly with the F protein, but, rather than inhibiting RSV entry into host cells, virus replication and budding, membrane-expressed RAGE or soluble RAGE blocked F-protein-mediated syncytium formation and sloughing. These data indicate that RAGE may contribute to protecting the lower airways from RSV by inhibiting the formation of syncytia, viral spread, epithelial damage and airway obstruction.

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The Human Respiratory Syncytial Virus Matrix Protein Is Required for Maturation of Viral Filaments

Journal of Virology ◽

10.1128/jvi.06744-11 ◽

2012 ◽

Vol 86 (8) ◽

pp. 4432-4443 ◽

Cited By ~ 44

Author(s):

Ruchira Mitra ◽

Pradyumna Baviskar ◽

Rebecca R. Duncan-Decocq ◽

Darshna Patel ◽

Antonius G. P. Oomens

Keyword(s):

Respiratory Syncytial Virus ◽

Matrix Protein ◽

Human Respiratory Syncytial Virus ◽

Syncytial Virus

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Association of the Matrix Protein from Respiratory Syncytial Virus with Lipid Monolayers and Bilayers

Biophysical Journal ◽

10.1016/j.bpj.2009.12.2641 ◽

2010 ◽

Vol 98 (3) ◽

pp. 485a

Author(s):

John M. Sanderson ◽

Helen K. McPhee ◽

Andrew Beeby ◽

Scott M.D. Watson

Keyword(s):

Respiratory Syncytial Virus ◽

Matrix Protein ◽

Lipid Monolayers ◽

The Matrix ◽

Syncytial Virus

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Structural Analysis of Respiratory Syncytial Virus Reveals the Position of M2-1 between the Matrix Protein and the Ribonucleoprotein Complex

Journal of Virology ◽

10.1128/jvi.00256-14 ◽

2014 ◽

Vol 88 (13) ◽

pp. 7602-7617 ◽

Cited By ~ 55

Author(s):

G. Kiss ◽

J. M. Holl ◽

G. M. Williams ◽

E. Alonas ◽

D. Vanover ◽

...

Keyword(s):

Respiratory Syncytial Virus ◽

Structural Analysis ◽

Matrix Protein ◽

Ribonucleoprotein Complex ◽

The Matrix ◽

Syncytial Virus

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Multiple, Non-conserved, Internal Viral Ligands Naturally Presented by HLA-B27 in Human Respiratory Syncytial Virus-infected Cells

Molecular & Cellular Proteomics ◽

10.1074/mcp.m900508-mcp200 ◽

2010 ◽

Vol 9 (7) ◽

pp. 1533-1539 ◽

Cited By ~ 18

Author(s):

Susana Infantes ◽

Elena Lorente ◽

Eilon Barnea ◽

Ilan Beer ◽

Juan José Cragnolini ◽

...

Keyword(s):

Respiratory Syncytial Virus ◽

Human Respiratory Syncytial Virus ◽

Hla B27 ◽

Infected Cells ◽

Syncytial Virus

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The Respiratory Syncytial Virus Matrix Protein Possesses a Crm1-Mediated Nuclear Export Mechanism

Journal of Virology ◽

10.1128/jvi.02374-08 ◽

2009 ◽

Vol 83 (11) ◽

pp. 5353-5362 ◽

Cited By ~ 56

Author(s):

Reena Ghildyal ◽

Adeline Ho ◽

Manisha Dias ◽

Lydia Soegiyono ◽

Phillip G. Bardin ◽

...

Keyword(s):

Respiratory Syncytial Virus ◽

Nuclear Export ◽

Matrix Protein ◽

Virus Assembly ◽

Nuclear Export Signal ◽

M Protein ◽

Nuclear Accumulation ◽

Infected Cells ◽

Syncytial Virus ◽

In Cells

ABSTRACT The respiratory syncytial virus (RSV) matrix (M) protein is localized in the nucleus of infected cells early in infection but is mostly cytoplasmic late in infection. We have previously shown that M localizes in the nucleus through the action of the importin β1 nuclear import receptor. Here, we establish for the first time that M's ability to shuttle to the cytoplasm is due to the action of the nuclear export receptor Crm1, as shown in infected cells, and in cells transfected to express green fluorescent protein (GFP)-M fusion proteins. Specific inhibition of Crm1-mediated nuclear export by leptomycin B increased M nuclear accumulation. Analysis of truncated and point-mutated M derivatives indicated that Crm1-dependent nuclear export of M is attributable to a nuclear export signal (NES) within residues 194 to 206. Importantly, inhibition of M nuclear export resulted in reduced virus production, and a recombinant RSV carrying a mutated NES could not be rescued by reverse genetics. That this is likely to be due to the inability of a nuclear export deficient M to localize to regions of virus assembly is indicated by the fact that a nuclear-export-deficient GFP-M fails to localize to regions of virus assembly when expressed in cells infected with wild-type RSV. Together, our data suggest that Crm1-dependent nuclear export of M is central to RSV infection, representing the first report of such a mechanism for a paramyxovirus M protein and with important implications for related paramyxoviruses.

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Downregulation of A20 Expression Increases the Immune Response and Apoptosis and Reduces Virus Production in Cells Infected by the Human Respiratory Syncytial Virus

Vaccines ◽

10.3390/vaccines8010100 ◽

2020 ◽

Vol 8 (1) ◽

pp. 100

Author(s):

María Martín-Vicente ◽

Rubén González-Sanz ◽

Isabel Cuesta ◽

Sara Monzón ◽

Salvador Resino ◽

...

Keyword(s):

Immune Response ◽

Respiratory Syncytial Virus ◽

Virus Production ◽

Antiviral Response ◽

Human Respiratory Syncytial Virus ◽

Negative Regulator ◽

Infected Cells ◽

Syncytial Virus ◽

Tract Infections ◽

The Impact

Human respiratory syncytial virus (HRSV) causes severe lower respiratory tract infections in infants, the elderly, and immunocompromised adults. Regulation of the immune response against HRSV is crucial to limiting virus replication and immunopathology. The A20/TNFAIP3 protein is a negative regulator of nuclear factor kappa B (NF-κB) and interferon regulatory factors 3/7 (IRF3/7), which are key transcription factors involved in the inflammatory/antiviral response of epithelial cells to virus infection. Here, we investigated the impact of A20 downregulation or knockout on HRSV growth and the induction of the immune response in those cells. Cellular infections in which the expression of A20 was silenced by siRNAs or eliminated by gene knockout showed increased inflammatory/antiviral response and reduced virus production. Similar results were obtained when the expression of A20-interacting proteins, such as TAX1BP1 and ABIN1, was silenced. Additionally, downregulation of A20, TAX1BP1, and ABIN1 increased cell apoptosis in HRSV-infected cells. These results show that the downregulation of A20 expression might contribute in the control of HRSV infections by potentiating the early innate immune response and increasing apoptosis in infected cells.

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