Identification of RSV Fusion Protein Interaction Domains on the Virus Receptor, Nucleolin
Nucleolin is an essential cellular receptor to human respiratory syncytial virus (RSV). Pharmacological targeting of the nucleolin RNA binding domain RBD1,2 can inhibit RSV infections in vitro and in vivo; however, the site(s) on RBD1,2 which interact with RSV are not known. We undertook a series of...
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doaj-36c366b2e51c4228944868cea69fc4e52021-02-09T00:04:36ZengMDPI AGViruses1999-49152021-02-011326126110.3390/v13020261Identification of RSV Fusion Protein Interaction Domains on the Virus Receptor, NucleolinPeter Mastrangelo0Allysia A. Chin1Stephanie Tan2Amy H. Jeon3Cameron A. Ackerley4Karen K. Siu5Jeffrey E. Lee6Richard G. Hegele7Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, CanadaDepartment of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, CanadaDepartment of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, CanadaDepartment of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, CanadaDepartment of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, CanadaDepartment of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, CanadaDepartment of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, CanadaDepartment of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S 1A8, CanadaNucleolin is an essential cellular receptor to human respiratory syncytial virus (RSV). Pharmacological targeting of the nucleolin RNA binding domain RBD1,2 can inhibit RSV infections in vitro and in vivo; however, the site(s) on RBD1,2 which interact with RSV are not known. We undertook a series of experiments designed to: document RSV-nucleolin co-localization on the surface of polarized MDCK cells using immunogold electron microscopy, to identify domains on nucleolin that physically interact with RSV using biochemical methods and determine their biological effects on RSV infection in vitro, and to carry out structural analysis toward informing future RSV drug development. Results of immunogold transmission and scanning electron microscopy showed RSV-nucleolin co-localization on the cell surface, as would be expected for a viral receptor. RSV, through its fusion protein (RSV-F), physically interacts with RBD1,2 and these interactions can be competitively inhibited by treatment with Palivizumab or recombinant RBD1,2. Treatment with synthetic peptides derived from two 12-mer domains of RBD1,2 inhibited RSV infection in vitro, with structural analysis suggesting these domains are potentially feasible for targeting in drug development. In conclusion, the identification and characterization of domains of nucleolin that interact with RSV provide the essential groundwork toward informing design of novel nucleolin-targeting compounds in RSV drug development.https://www.mdpi.com/1999-4915/13/2/261RSVnucleolinfusion proteinprotein–protein interactions |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Peter Mastrangelo Allysia A. Chin Stephanie Tan Amy H. Jeon Cameron A. Ackerley Karen K. Siu Jeffrey E. Lee Richard G. Hegele |
spellingShingle |
Peter Mastrangelo Allysia A. Chin Stephanie Tan Amy H. Jeon Cameron A. Ackerley Karen K. Siu Jeffrey E. Lee Richard G. Hegele Identification of RSV Fusion Protein Interaction Domains on the Virus Receptor, Nucleolin Viruses RSV nucleolin fusion protein protein–protein interactions |
author_facet |
Peter Mastrangelo Allysia A. Chin Stephanie Tan Amy H. Jeon Cameron A. Ackerley Karen K. Siu Jeffrey E. Lee Richard G. Hegele |
author_sort |
Peter Mastrangelo |
title |
Identification of RSV Fusion Protein Interaction Domains on the Virus Receptor, Nucleolin |
title_short |
Identification of RSV Fusion Protein Interaction Domains on the Virus Receptor, Nucleolin |
title_full |
Identification of RSV Fusion Protein Interaction Domains on the Virus Receptor, Nucleolin |
title_fullStr |
Identification of RSV Fusion Protein Interaction Domains on the Virus Receptor, Nucleolin |
title_full_unstemmed |
Identification of RSV Fusion Protein Interaction Domains on the Virus Receptor, Nucleolin |
title_sort |
identification of rsv fusion protein interaction domains on the virus receptor, nucleolin |
publisher |
MDPI AG |
series |
Viruses |
issn |
1999-4915 |
publishDate |
2021-02-01 |
description |
Nucleolin is an essential cellular receptor to human respiratory syncytial virus (RSV). Pharmacological targeting of the nucleolin RNA binding domain RBD1,2 can inhibit RSV infections in vitro and in vivo; however, the site(s) on RBD1,2 which interact with RSV are not known. We undertook a series of experiments designed to: document RSV-nucleolin co-localization on the surface of polarized MDCK cells using immunogold electron microscopy, to identify domains on nucleolin that physically interact with RSV using biochemical methods and determine their biological effects on RSV infection in vitro, and to carry out structural analysis toward informing future RSV drug development. Results of immunogold transmission and scanning electron microscopy showed RSV-nucleolin co-localization on the cell surface, as would be expected for a viral receptor. RSV, through its fusion protein (RSV-F), physically interacts with RBD1,2 and these interactions can be competitively inhibited by treatment with Palivizumab or recombinant RBD1,2. Treatment with synthetic peptides derived from two 12-mer domains of RBD1,2 inhibited RSV infection in vitro, with structural analysis suggesting these domains are potentially feasible for targeting in drug development. In conclusion, the identification and characterization of domains of nucleolin that interact with RSV provide the essential groundwork toward informing design of novel nucleolin-targeting compounds in RSV drug development. |
topic |
RSV nucleolin fusion protein protein–protein interactions |
url |
https://www.mdpi.com/1999-4915/13/2/261 |
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