Parvovirus infection of cells by using variants of the feline transferrin receptor altering clathrin-mediated endocytosis, membrane domain localization, and capsid-binding domains.
The feline and canine transferrin receptors (TfRs) bind canine parvovirus to host cells and mediate rapid capsid uptake and infection. The TfR and its ligand transferrin have well-described pathways of endocytosis and recycling. Here we tested several receptor-dependent steps in infection for their role in virus infection of cells. Deletions of cytoplasmic sequences or mutations of the Tyr-Thr-Arg-Phe internalization motif reduced the rate of receptor uptake from the cell surface, while polar residues introduced into the transmembrane sequence resulted in increased degradation of transferrin. However, the mutant receptors still mediated efficient virus infection. In contrast, replacing the cytoplasmic and transmembrane sequences of the feline TfR with those of the influenza virus neuraminidase (NA) resulted in a receptor that bound and endocytosed the capsid but did not mediate viral infection. This chimeric receptor became localized to detergent-insoluble membrane domains. To test the effect of structural virus receptor interaction on infection, two chimeric receptors were prepared which contained antibody-variable domains that bound the capsid in place of the TfR ectodomain. These chimeric receptors bound CPV capsids and mediated uptake but did not result in cell infection. Adding soluble feline TfR ectodomain to the virus during that uptake did not allow infection.
Hueffer
Karsten
K
James A. Baker Institute for Animal Health, Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
Palermo
Laura M
LM
Parrish
Colin R
CR
D000595:Amino Acid Sequence; D000818:Animals; D001666:Binding Sites, Antibody; D002213:Capsid / Q000378:metabolism*; D002415:Cats; D002462:Cell Membrane / Q000378:metabolism*; D002966:Clathrin / Q000502:physiology*; D004705:Endocytosis; D028301:Feline Panleukopenia Virus / Q000502:physiology*; D008969:Molecular Sequence Data; D011990:Receptors, Transferrin / Q000502:physiology*; D011991:Receptors, Virus / Q000502:physiology*
D002966:Clathrin; D011990:Receptors, Transferrin; D011991:Receptors, Virus