Two retroviral entry pathways distinguished by lipid raft association of the viral receptor and differences in viral infectivity.
The receptor "priming" model for entry of the retrovirus avian sarcoma and leukosis virus (ASLV) predicts that upon binding cell surface receptors, virions are endocytosed and trafficked to acidic endosomes where fusion occurs. To test this model directly, we have now followed subgroup A ASLV (ASLV-A) virions entering cells via either the transmembrane (TVA950) or glycophosphatidylinositol (GPI)-anchored (TVA800) forms of the cellular receptor. Our results suggest that viruses entering via these two forms of receptor are subjected to different intracellular fates, perhaps due to use of different endocytic trafficking pathways to access acidic fusion compartments. Kinetic analyses demonstrated that virus bound to TVA800 was taken up from the cell surface more slowly but then trafficked to the site of fusion more quickly than that entering via TVA950. Furthermore, transiently arresting virions within putative fusion compartments with NH4Cl led to a substantially greater decrease in the infectivity of virions using TVA950 than with those using TVA800. The increased infectivity of virions using TVA800 correlated with the localization of this receptor to lipid rafts, since this effect was abolished by pharmacological disruption of lipid rafts. Together these results suggest that, in the presence of NH4Cl, virus bound to the GPI-anchored receptor may utilize a lipid raft-dependent pathway to accumulate within a fusion compartment where it is more stable than if it enters via the transmembrane receptor. The TVA800/ASLV-A system should prove useful for the molecular analysis of lipid raft-dependent endocytosis and may provide a tool for the biochemical dissection of the poorly understood uncoating step of retroviral replication.
Narayan
Shakti
S
Cellular and Molecular Biology Program, Department of Oncology, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
Barnard
Richard J O
RJ
Young
John A T
JA
D001355:Alpharetrovirus / Q000502:physiology*; D000643:Ammonium Chloride / Q000494:pharmacology; D002460:Cell Line; D003505:Cyclodextrins / Q000494:pharmacology; D004705:Endocytosis; D006801:Humans; D021962:Membrane Microdomains / Q000502:physiology*; D011991:Receptors, Virus / Q000502:physiology*; D014771:Virion / Q000502:physiology
D003505:Cyclodextrins; D011991:Receptors, Virus; D000643:Ammonium Chloride