@article{132451,
  keywords = {Animals, Humans, Rats, Cell Line, Protein Interaction Mapping, Fibroblasts, Viral Nonstructural Proteins, Sindbis Virus},
  author = {Ileana Cristea and John-William Carroll and Michael Rout and Charles Rice and Brian Chait and Margaret MacDonald},
  title = {Tracking and elucidating alphavirus-host protein interactions},
  abstract = {<p>Viral infections cause profound alterations in host cells. Here, we explore the interactions between proteins of the Alphavirus Sindbis and host factors during the course of mammalian cell infection. Using a mutant virus expressing the viral nsP3 protein tagged with green fluorescent protein (GFP) we directly observed nsP3 localization and isolated nsP3-interacting proteins at various times after infection. These results revealed that host factor recruitment to nsP3-containing complexes was time dependent, with a specific early and persistent recruitment of G3BP and a later recruitment of 14-3-3 proteins. Expression of GFP-tagged G3BP allowed reciprocal isolation of nsP3 in Sindbis infected cells, as well as the identification of novel G3BP-interacting proteins in both uninfected and infected cells. Note-worthy interactions include nuclear pore complex components whose interactions with G3BP were reduced upon Sindbis infection. This suggests that G3BP is a nuclear transport factor, as hypothesized previously, and that viral infection may alter RNA transport. Immunoelectron microscopy showed that a portion of Sindbis nsP3 is localized at the nuclear envelope, suggesting a possible site of G3BP recruitment to nsP3-containing complexes. Our results demonstrate the utility of using a standard GFP tag to both track viral protein localization and elucidate specific viral-host interactions over time in infected mammalian cells.</p>
},
  year = {2006},
  journal = {J Biol Chem},
  volume = {281},
  pages = {30269-78},
  month = {10/2006},
  issn = {0021-9258},
  doi = {10.1074/jbc.M603980200},
  language = {eng},
}