Intracellular topology and epitope shielding of poliovirus 3A protein.
The poliovirus RNA replication complex comprises multiple viral and possibly cellular proteins assembled on the cytoplasmic surface of rearranged intracellular membranes. Viral proteins 3A and 3AB perform several functions during the poliovirus replicative cycle, including significant roles in rearranging membranes, anchoring the viral polymerase to these membranes, inhibiting host protein secretion, and possibly providing the 3B protein primer for RNA synthesis. During poliovirus infection, the immunofluorescence signal of an amino-terminal epitope of 3A-containing proteins is markedly shielded compared to 3A protein expressed in the absence of other poliovirus proteins. This is not due to luminal orientation of all or a subset of the 3A-containing polypeptides, as shown by immunofluorescence following differential permeabilization and proteolysis experiments. Shielding of the 3A epitope is more pronounced in cells infected with wild-type poliovirus than in cells with temperature-sensitive mutant virus that contains a mutation in the 3D polymerase coding region adjacent to the 3AB binding site. Therefore, it is likely that direct binding of the poliovirus RNA-dependent RNA polymerase occludes the amino terminus of 3A-containing polypeptides in the RNA replication complex.
Choe
Sunny S
SS
299 Campus Dr., Stanford University School of Medicine, Stanford, CA 94305, USA.
Kirkegaard
Karla
K
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D000939:Epitopes; D019084:Fluorescent Antibody Technique, Indirect; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000096:biosynthesis; D012324:RNA-Dependent RNA Polymerase / Q000378:metabolism; D014162:Transfection; D014758:Viral Core Proteins / Q000737:chemistry* / Q000276:immunology / Q000502:physiology
C109349:3A protein, Poliovirus; D000939:Epitopes; D012367:RNA, Viral; D014758:Viral Core Proteins; D012324:RNA-Dependent RNA Polymerase