title: Equine Rhinitis A Virus Mutants with Altered Acid Resistance Unveil a Key Role of VP3 and Intrasubunit Interactions in the Control of the pH Stability of the Aphthovirus Capsid.
abstract: Equine rhinitis A virus (ERAV) is a picornavirus associated with respiratory disease in horses and is genetically closely related to foot-and-mouth disease virus (FMDV), the prototype aphthovirus. ERAV has recently gained interest as an FMDV alternative for the study of aphthovirus biology, including cell entry and uncoating or antiviral testing. As described for FMDV, current data support that acidic pH inside cellular endosomes triggers ERAV uncoating. In order to provide further insights into aphthovirus uncoating mechanism, we have isolated a panel of ERAV mutants with altered acid sensitivity and that differed on their degree of sensitivity to the inhibition of endosome acidification. These results provide functional evidence of the involvement of acidic pH on ERAV uncoating within endosomes. Remarkably, all amino acid substitutions found in acid-labile or acid-resistant ERAVs were located in the capsid protein VP3, indicating that this protein plays a pivotal role for the control of pH stability of the ERAV capsid. Moreover, all amino acid substitutions mapped at the intraprotomer interface between VP3 and VP2 or between VP3 and the N terminus of VP1. These results expand our knowledge on the regions that regulate the acid stability of aphthovirus capsid and should be taken into account when using ERAV as a surrogate of FMDV.
  
  
  OBJECTIVE
  The viral capsid constitutes a sort of dynamic nanomachine that protects the viral genome against environmental assaults while accomplishing important functions such as receptor attachment for viral entry or genome release. We have explored the molecular determinants of aphthovirus capsid stability by isolating and characterizing a panel of equine rhinitis A virus mutants that differed on their acid sensitivity. All the mutations were located within a specific region of the capsid, the intraprotomer interface among capsid proteins, thus providing new insights into the regions that control the acid stability of aphthovirus capsid. These findings could positively contribute to the development of antiviral approaches targeting aphthovirus uncoating or the refinement of vaccine strategies based on capsid stabilization.
authors.affiliation: Department of Virology and Microbiology, Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Madrid, Spain.
authors.forename: Flavia
authors.initials: F
authors.lastname: Caridi
authors.affiliation: Department of Virology and Microbiology, Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Madrid, Spain.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Cañas-Arranz
authors.affiliation: Department of Virology and Microbiology, Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Madrid, Spain.
authors.forename: Angela
authors.initials: A
authors.lastname: Vázquez-Calvo
authors.affiliation: Department of Virology and Microbiology, Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Madrid, Spain fsobrino@cbm.csic.es.
authors.forename: Francisco
authors.initials: F
authors.lastname: Sobrino
authors.affiliation: Department of Virology and Microbiology, Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Madrid, Spain.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Martín-Acebes
mesh_terms: D000143:Acids / Q000378:metabolism*; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D005537:Aphthovirus / Q000187:drug effects / Q000235:genetics*; D002213:Capsid / Q000187:drug effects; D036022:Capsid Proteins / Q000235:genetics*; D011992:Endosomes / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000187:drug effects; D016679:Genome, Viral / Q000235:genetics; D006736:Horses / Q000821:virology*; D006863:Hydrogen-Ion Concentration; D009154:Mutation / Q000235:genetics; D010850:Picornaviridae Infections / Q000188:drug therapy / Q000821:virology; D053586:Virus Internalization / Q000187:drug effects
chemical_list: D000143:Acids; D000998:Antiviral Agents; D036022:Capsid Proteins; C442460:VP3 protein, Foot-and-mouth disease virus