title: Membrane-destabilizing activity of rotavirus NSP4 is mediated by a membrane-proximal amphipathic domain.
abstract: Expression of the rotavirus non-structural glycoprotein NSP4 in E. coli leads to a decrease in optical density of the culture and release of [(3)H]uridine into the medium, effects attributable to the ability of NSP4 to perturb the bacterial membrane. To identify a domain of NSP4 responsible, different regions of the polypeptide were expressed in E. coli. Membrane destabilization is associated with a region of the protein located within residues 48-91, which includes a potential cationic amphipathic helix. A second region of NSP4 that contains a coiled-coil oligomerization domain and a sequence reported to function as a viral enterotoxin enhances the membrane-destabilizing activity of residues 48-91, but has no direct effect on the membrane stability. These studies suggest that the membrane-destabilizing and enterotoxic properties of NSP4 may be mediated by different regions of the polypeptide and suggest a possible basis for the cytotoxicity of NSP4 in mammalian cells.
authors.affiliation: Microbiology and Virology Research Group, School of Biological Sciences, University of Auckland, Private Bag 92019, Auckland, New Zealand1.
authors.forename: Edward P
authors.initials: EP
authors.lastname: Browne
authors.affiliation: Microbiology and Virology Research Group, School of Biological Sciences, University of Auckland, Private Bag 92019, Auckland, New Zealand1.
authors.forename: A Richard
authors.initials: AR
authors.lastname: Bellamy
authors.affiliation: Microbiology and Virology Research Group, School of Biological Sciences, University of Auckland, Private Bag 92019, Auckland, New Zealand1.
authors.forename: John A
authors.initials: JA
authors.lastname: Taylor
mesh_terms: D002463:Cell Membrane Permeability / Q000187:drug effects; D012321:DNA-Directed RNA Polymerases; D004721:Endoplasmic Reticulum / Q000502:physiology; D004926:Escherichia coli / Q000187:drug effects; D017433:Protein Structure, Secondary; D012401:Rotavirus / Q000737:chemistry*; D014529:Uridine / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000633:toxicity
chemical_list: D017361:Viral Nonstructural Proteins; D012321:DNA-Directed RNA Polymerases; D014529:Uridine