Fusion-defective gibbon ape leukemia virus vectors can be rescued by homologous but not heterologous soluble envelope proteins.
Murine leukemia virus (MLV)-derived envelope proteins containing alterations in or adjacent to the highly conserved PHQ motif present at the N terminus of the envelope surface subunit (SU) are incorporated into vector particles but are not infectious due to a postbinding block to viral entry. These mutants can be rendered infectious by the addition of soluble receptor-binding domain (RBD) proteins in the culture medium. The RBD proteins that rescue the infectivity of these defective MLV vectors can be derived from the same MLV or from other MLVs that use distinct receptors to mediate entry. We have now constructed functional immunologically reactive gibbon ape leukemia virus (GALV) envelope proteins, tagged with a feline leukemia virus (FeLV)-derived epitope tag, which are efficiently incorporated into infectious particles. Tagged GALV envelope proteins bind specifically to cells expressing the phosphate transporter protein Pit1, demonstrating for the first time that Pit1 is the binding receptor for GALV and not a coreceptor or another type of GALV entry factor. We have also determined that GALV particles bearing SU proteins with an insertion C-terminal to the PHQ motif (GALV I(10)) bind Pit1 but fail to infect cells. Incubation with soluble GALV RBD renders GALV I(10) particles infectious, whereas incubation with soluble RBDs from MLV or FeLV-B does not. This finding is consistent with the results obtained by Lauring et al. using FeLV-T, a virus that employs Pit1 as a receptor but requires soluble FeLV RBD for entry. MLV and GALV RBDs are not able to render FeLV-T infectious (A. S. Lauring, M. M. Anderson, and J. Overbaugh, J. Virol. 75:8888-8898, 2001). Together, these results suggest that fusion-defective FeLV-T and GALV are restricted to homologous RBD rescue of infectivity.
Farrell
Karen B
KB
Unit on Molecular Virology, Laboratory of Cellular and Molecular Regulation, National Institute of Mental Health, Bethesda, Maryland 20892, USA.
Ting
Yuan-Tsang
YT
Eiden
Maribeth V
MV
D000595:Amino Acid Sequence; D000818:Animals; D018845:CD11 Antigens / Q000737:chemistry / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism; D002415:Cats; D002460:Cell Line; D003673:Defective Viruses; D000939:Epitopes; D005822:Genetic Vectors; D005256:Leukemia Virus, Feline / Q000235:genetics / Q000378:metabolism; D017989:Leukemia Virus, Gibbon Ape / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D008561:Membrane Fusion; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D049294:Phospholipid Transfer Proteins; D012995:Solubility; D013045:Species Specificity; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*
D018845:CD11 Antigens; D002352:Carrier Proteins; D000939:Epitopes; D008565:Membrane Proteins; D049294:Phospholipid Transfer Proteins; D014759:Viral Envelope Proteins