title: Budding of equine infectious anemia virus is insensitive to proteasome inhibitors.
abstract: The only retrovirus protein required for the budding of virus-like particles is the Gag protein; however, recent studies of Rous sarcoma virus (RSV) and human immunodeficiency virus have suggested that modification of Gag with ubiquitin (Ub) is also required. As a consequence, the release of these viruses is reduced in the presence of proteasome inhibitors, which indirectly reduce the levels of free Ub within the cell. Here we show that the budding of equine infectious anemia virus (EIAV) from infected equine cells is largely unaffected by these drugs, although use of one inhibitor (MG-132) resulted in a dramatic block to proteolytic processing of Gag. This lack of sensitivity was also observed in transiently transfected avian cells under conditions that greatly reduce RSV budding. Moreover, insensitivity was observed when the EIAV Gag protein was expressed in the absence of all the other virus products, indicating that they are not required for this phenotype. An activity that enables EIAV to tolerate exposure to proteasome inhibitors was mapped to the C-terminal p9 sequence, as demonstrated by the ability of an RSV Gag-p9 chimera to bud in the presence of the drugs. Intriguingly, the p9 sequence contains a short sequence motif that is similar to a surface-exposed helix of Ub, suggesting that EIAV Gag may have captured a function that allows it to bypass the need for ubiquitination. Thus, the mechanism of EIAV budding may not be substantially different from that of other retroviruses, even though it behaves differently in the presence of proteasome inhibitors.
authors.affiliation: Department of Microbiology and Immunology, The Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033, USA.
authors.forename: Akash
authors.initials: A
authors.lastname: Patnaik
authors.forename: Vincent
authors.initials: V
authors.lastname: Chau
authors.forename: Feng
authors.initials: F
authors.lastname: Li
authors.forename: Ronald C
authors.initials: RC
authors.lastname: Montelaro
authors.forename: John W
authors.initials: JW
authors.lastname: Wills
mesh_terms: D000818:Animals; D002460:Cell Line; D003546:Cysteine Endopeptidases; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D015683:Gene Products, gag / Q000378:metabolism; D004860:Infectious Anemia Virus, Equine / Q000187:drug effects* / Q000235:genetics / Q000502:physiology*; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors*; D046988:Proteasome Endopeptidase Complex; D014162:Transfection; D014452:Ubiquitins / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D015683:Gene Products, gag; D009097:Multienzyme Complexes; D014452:Ubiquitins; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex