Characterization of essential domains and plasticity of the classical Swine Fever virus Core protein.
Pestiviruses are pathogens of cloven-hoofed animals, belonging to the Flaviviridae. The pestiviral particle consists of a lipid membrane containing the three envelope glycoproteins Erns, E1, and E2 and a nucleocapsid of unknown symmetry, which is composed of the Core protein and the viral positive-sense RNA genome. The positively charged pestiviral Core protein consists of 86 to 89 amino acids. To analyze the organization of essential domains, N- and C-terminal truncations, as well as internal deletions, were introduced into the Core coding sequence in the context of an infectious cDNA clone of classical swine fever virus strain Alfort. Amino acids 179 to 180, 194 to 198, and 208 to 212 proved to be of special importance for the generation of progeny virus. The results of transcomplementation of a series of C-terminally truncated Core molecules indicate the importance of Ala255 at the C terminus. The plasticity of Core protein was examined by the construction of concatemeric arrays of Core coding regions and the insertion of up to three yellow fluorescent protein (YFP) genes between two Core genes. Even a Core fusion protein with more than 10-fold-increased molecular mass was integrated into the viral particle and supported the production of infectious progeny virus. The unexpected plasticity of Core protein brings into question the formation of a regular icosahedric particle and supports the idea of a histone-like protein-RNA interaction. All viruses with a duplicated Core gene were unstable and reverted to the wild-type sequence. Interestingly, a nonviable YFP-Core construct was rescued by a mutation within the C-terminal domain of the nonstructural protein NS3.
Riedel
Christiane
C
Institute of Virology, Faculty of Veterinary Medicine, Justus-Liebig University Giessen, Frankfurter Strasse 107, 35392 Giessen, Germany.
Lamp
Benjamin
B
Heimann
Manuela
M
Rümenapf
Till
T
D000595:Amino Acid Sequence; D000596:Amino Acids; D000818:Animals; D006692:Classical Swine Fever Virus; D018076:DNA, Complementary; D009154:Mutation; D013552:Swine; D014758:Viral Core Proteins / Q000235:genetics; D014771:Virion
D000596:Amino Acids; D018076:DNA, Complementary; D014758:Viral Core Proteins