Recombinant Dengue virus protein NS2B alters membrane permeability in different membrane models
Top Cited Papers
Open Access
- 4 January 2016
- journal article
- research article
- Published by Springer Science and Business Media LLC in Virology Journal
- Vol. 13 (1), 1-11
- https://doi.org/10.1186/s12985-015-0456-4
Abstract
One of the main phenomena occurring in cellular membranes during virus infection is a change in membrane permeability. It has been observed that numerous viral proteins can oligomerize and form structures known as viroporins that alter the permeability of membranes. Previous findings have identified such proteins in cells infected with Japanese encephalitis virus (JEV), a member of the same family that Dengue virus (DENV) belongs to (Flaviviridae). In the present work, we investigated whether the small hydrophobic DENV protein NS2B serves a viroporin function. We cloned the DENV NS2B sequence and expressed it in a bacterial expression system. Subsequently, we evaluated the effect of DENV NS2B on membranes when NS2B was overexpressed, measured bacterial growth restriction, and evaluated changes of permeability to hygromycin. The NS2B protein was purified by affinity chromatography, and crosslinking assays were performed to determine the presence of oligomers. Hemolysis assays and transmission electron microscopy were performed to identify structures involved in permeability changes. The DENV-2 NS2B protein showed similitude with the JEV viroporin. The DENV-2 NS2B protein possessed the ability to change the membrane permeability in bacteria, to restrict bacterial cell growth, and to enable membrane permeability to hygromycin B. The NS2B protein formed trimers that could participate in cell lysis and generate organized structures on eukaryotes membranes. Our data suggest that the DENV-2 NS2B viral protein is capable of oligomerizing and organizing to form pore-like structures in different lipid environments, thereby modifying the permeability of cell membranes.Keywords
Funding Information
- Consejo Nacional de Ciencia y Tecnología (MX) (897)
- CONACyT (CB‐2010‐01/0154270)
This publication has 37 references indexed in Scilit:
- Early Dengue Virus Protein Synthesis Induces Extensive Rearrangement of the Endoplasmic Reticulum Independent of the UPR and SREBP-2 PathwayPLOS ONE, 2012
- High-Risk Human Papillomavirus E5 Oncoprotein Displays Channel-Forming Activity Sensitive to Small-Molecule InhibitorsJournal of Virology, 2012
- Activation of the Innate Immune Response against DENV in Normal Non-Transformed Human FibroblastsPLoS Neglected Tropical Diseases, 2011
- The SV40 Late Protein VP4 Is a Viroporin that Forms Pores to Disrupt Membranes for Viral ReleasePLoS Pathogens, 2011
- Influenza M2 proton channelsBiochimica et Biophysica Acta (BBA) - Biomembranes, 2011
- Rotavirus Disrupts Calcium Homeostasis by NSP4 Viroporin ActivitymBio, 2010
- Direct visualization of the small hydrophobic protein of human respiratory syncytial virus reveals the structural basis for membrane permeabilityFEBS Letters, 2010
- The Human Polyoma JC Virus Agnoprotein Acts as a ViroporinPLoS Pathogens, 2010
- The 3-dimensional structure of a hepatitis C virus p7 ion channel by electron microscopyProceedings of the National Academy of Sciences of the United States of America, 2009
- Composition and Three-Dimensional Architecture of the Dengue Virus Replication and Assembly SitesCell Host & Microbe, 2009