THE EQUINE HERPESVIRUS-1 IR6 PROTEIN THAT COLOCALIZES WITH NUCLEAR LAMINS IS INVOLVED IN NUCLEOCAPSID EGRESS AND MIGRATES FROM CELL-TO-CELLINDEPENDENTLY OF VIRUS-INFECTION

The equine herpesvirus 1 (EW-I) IR6 protein forms typical rod-like str uctures in infected cells, influences virus growth at elevated tempera tures, and determines the virulence of EHV-I Rac strains (Osterrieder et al., Virology 226:243-251, 1996). Experiments to further elucidate the functions and properties of the IR6 protein were conducted. It was shown that the IR6 protein of wild-type RacL11 virus colocalizes with nuclear lamins very late in infection as demonstrated by confocal las er scan microscopy and coimmunoprecipitation experiments. In contrast, the mutated IR6 protein encoded by the RacM24 strain did not colocali ze with the Iamin proteins at any time postinfection (p.i.). Electron microscopical examinations of ultrathin sections were performed on cel ls infected at 37 and 40 degrees C, the latter being a temperature at which the IR6-negative RacH virus and the RacM24 virus are greatly imp aired in virus replication. These analyses revealed that nucleocapsid formation is efficient at 40 degrees C irrespective of the virus strai n. However, whereas cytoplasmic virus particles were readily observed at 16 h p.i. in cells infected with the wild-type EHV-1 RacL11 or an I R6-recombinant RacH virus (HIR6-1) at 40 degrees C, virtually no capsi d translocation to the cytoplasm was obvious in RacH- or RacM24-infect ed cells at the elevated temperature, demonstrating that the IR6 prote in is involved in nucleocapsid egress. Transient transfection assays u sing RacL11 or RacM24 IR6 plasmid DNA and COS7 or Rk(13) cells, infect ion studies using a gB-negative RacL11 mutant (L11 Delta gB) which is deficient in direct cell-to-cell spread, and studies using lysates of IR6-transfected cells demonstrated that the wild-type IR6 protein is t ransported from cell to cell in the absence of virus infection and can enter cells by a yet unknown mechanism.