title: Recombinant Dengue virus protein NS2B alters membrane permeability in different membrane models.
abstract: BACKGROUND
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Departmento de Biomedicina Molecular, Centro de Investigacion y Estudios avanzados IPN, Av. Instituto Politecnico 2508 Col. San Pedro Zacatenco, 07360, México, Mexico. mleonj@cinvestav.mx.|Present Address: Departamento de Inmunobioquimica, Instituto Nacional de Perinatología, Montes Urales #800 Col. Lomas de Virreyes, 1100, México, Mexico. mleonj@cinvestav.mx.
authors.forename: Moisés
authors.initials: M
authors.lastname: León-Juárez
authors.affiliation: Departmento de Biomedicina Molecular, Centro de Investigacion y Estudios avanzados IPN, Av. Instituto Politecnico 2508 Col. San Pedro Zacatenco, 07360, México, Mexico. correommc58@gmail.com.
authors.forename: Macario
authors.initials: M
authors.lastname: Martínez-Castillo
authors.affiliation: Departmento de Biomedicina Molecular, Centro de Investigacion y Estudios avanzados IPN, Av. Instituto Politecnico 2508 Col. San Pedro Zacatenco, 07360, México, Mexico. gauravshrivastava01@gmail.com.
authors.forename: Gaurav
authors.initials: G
authors.lastname: Shrivastava
authors.affiliation: Departmento de Biomedicina Molecular, Centro de Investigacion y Estudios avanzados IPN, Av. Instituto Politecnico 2508 Col. San Pedro Zacatenco, 07360, México, Mexico. leebeydengue@yahoo.com.mx.
authors.forename: Julio
authors.initials: J
authors.lastname: García-Cordero
authors.affiliation: Departmento de Biomedicina Molecular, Centro de Investigacion y Estudios avanzados IPN, Av. Instituto Politecnico 2508 Col. San Pedro Zacatenco, 07360, México, Mexico. nvillega@cinvestav.mx.
authors.forename: Nicolás
authors.initials: N
authors.lastname: Villegas-Sepulveda
authors.affiliation: Departamento de Bioquímica, CINVESTAV IPN Av, IPN # 2508 Col. San Pedro Zacatenco, 07360, México, DF, Mexico. mmondrag@cinvestav.mx.
authors.forename: Mónica
authors.initials: M
authors.lastname: Mondragón-Castelán
authors.affiliation: Departamento de Bioquímica, CINVESTAV IPN Av, IPN # 2508 Col. San Pedro Zacatenco, 07360, México, DF, Mexico. rmflores@cinvestav.mx.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Mondragón-Flores
authors.affiliation: Departmento de Biomedicina Molecular, Centro de Investigacion y Estudios avanzados IPN, Av. Instituto Politecnico 2508 Col. San Pedro Zacatenco, 07360, México, Mexico. lcedillo@cinvestav.mx.
authors.forename: Leticia
authors.initials: L
authors.lastname: Cedillo-Barrón
mesh_terms: D000595:Amino Acid Sequence; D002463:Cell Membrane Permeability / Q000187:drug effects*; D003716:Dengue Virus / Q000235:genetics / Q000378:metabolism*; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000648:ultrastructure; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D011487:Protein Conformation; D055503:Protein Multimerization; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D016415:Sequence Alignment; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: D011993:Recombinant Fusion Proteins; D017361:Viral Nonstructural Proteins