Peptides Interfering 3A Protein Dimerization Decrease FMDV Multiplication.
Nonstructural protein 3A is involved in relevant functions in foot-and-mouth disease virus (FMDV) replication. FMDV 3A can form homodimers and preservation of the two hydrophobic α-helices (α1 and α2) that stabilize the dimer interface is essential for virus replication. In this work, small peptides mimicking residues involved in the dimer interface were used to interfere with dimerization and thus gain insight on its biological function. The dimer interface peptides α1, α2 and that spanning the two hydrophobic α-helices, α12, impaired in vitro dimer formation of a peptide containing the two α-helices, this effect being higher with peptide α12. To assess the effect of dimer inhibition in cultured cells, the interfering peptides were N-terminally fused to a heptaarginine (R7) sequence to favor their intracellular translocation. Thus, when fused to R7, interference peptides (100 μM) were able to inhibit dimerization of transiently expressed 3A, the higher inhibitions being found with peptides α1 and α12. The 3A dimerization impairment exerted by the peptides correlated with significant, specific reductions in the viral yield recovered from peptide-treated FMDV infected cells. In this case, α2 was the only peptide producing significant reductions at concentrations lower than 100 μM. Thus, dimer interface peptides constitute a tool to understand the structure-function relationship of this viral protein and point to 3A dimerization as a potential antiviral target.
Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Cantoblanco, Madrid, Spain.
Mónica
M
González-Magaldi
Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Cantoblanco, Madrid, Spain.
Ángela
Á
Vázquez-Calvo
Departament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, Barcelona, Spain.
Beatriz G
BG
de la Torre
Departament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, Barcelona, Spain.
Javier
J
Valle
Departament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, Barcelona, Spain.
David
D
Andreu
Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Cantoblanco, Madrid, Spain.
Francisco
F
Sobrino
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D005536:Foot-and-Mouth Disease / Q000235:genetics / Q000821:virology*; D029544:Foot-and-Mouth Disease Virus / Q000737:chemistry / Q000235:genetics*; D057927:Hydrophobic and Hydrophilic Interactions; D051379:Mice; D055503:Protein Multimerization; D017433:Protein Structure, Secondary; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000235:genetics; D014779:Virus Replication / Q000235:genetics*
D017361:Viral Nonstructural Proteins