Distinct intracellular trafficking of equine infectious anemia virus and human immunodeficiency virus type 1 Gag during viral assembly and budding revealed by bimolecular fluorescence complementation assays.
Retroviral Gag polyproteins are necessary and sufficient for virus budding. Numerous studies of human immunodeficiency virus type 1 (HIV-1) Gag assembly and budding mechanisms have been reported, but relatively little is known about these fundamental pathways among animal lentiviruses. While there may be a general assumption that lentiviruses share common assembly mechanisms, studies of equine infectious anemia virus (EIAV) have indicated alternative cellular pathways and cofactors employed among lentiviruses for assembly and budding. In the current study, we used bimolecular fluorescence complementation to characterize and compare assembly sites and budding efficiencies of EIAV and HIV-1 Gag in both human and rodent cells. The results of these studies demonstrated that replacing the natural RNA nuclear export element (Rev-response element [RRE]) used by HIV-1 and EIAV with the hepatitis B virus posttranscriptional regulatory element (PRE) altered HIV-1, but not EIAV, Gag assembly sites and budding efficiency in human cells. Consistent with this novel observation, different assembly sites were revealed in human cells for Rev-dependent EIAV and HIV-1 Gag polyproteins. In rodent cells, Rev-dependent HIV-1 Gag assembly and budding were blocked, but changing RRE to PRE rescued HIV-1 Gag assembly and budding. In contrast, EIAV Gag polyproteins synthesized from mRNA exported via either Rev-dependent or PRE-dependent mechanisms were able to assemble and bud efficiently in rodent cells. Taken together, our results suggest that lentivirus assembly and budding are regulated by the RNA nuclear export pathway and that alternative cellular pathways can be adapted for lentiviral Gag assembly and budding.
Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, W1144 Biomedical Science Tower, Pittsburgh, PA 15261, USA.
Jing
J
Jin
Timothy
T
Sturgeon
Chaoping
C
Chen
Simon C
SC
Watkins
Ora A
OA
Weisz
Ronald C
RC
Montelaro
D021581:Active Transport, Cell Nucleus; D000818:Animals; D002460:Cell Line; D005453:Fluorescence; D015683:Gene Products, gag / Q000378:metabolism*; D015694:Gene Products, rev / Q000502:physiology; D015497:HIV-1 / Q000378:metabolism / Q000502:physiology*; D006736:Horses; D006801:Humans; D004860:Infectious Anemia Virus, Equine / Q000378:metabolism / Q000502:physiology*; D021381:Protein Transport; D020218:Response Elements; D012377:Rodentia; D019065:Virus Assembly
D015683:Gene Products, gag; D015694:Gene Products, rev