       Document 0102
 DOCN  M9460102
 TI    Homolog-scanning mutagenesis reveals poliovirus receptor residues
       important for virus binding and replication.
 DT    9404
 AU    Morrison ME; He YJ; Wien MW; Hogle JM; Racaniello VR; Department of
       Microbiology, Columbia University College of; Physicians & Surgeons, New
       York, New York 10032.
 SO    J Virol. 1994 Apr;68(4):2578-88. Unique Identifier : AIDSLINE
       MED/94187099
 AB    Poliovirus initiates infection of primate cells by binding to the
       poliovirus receptor, Pvr. Mouse cells do not bind poliovirus but express
       a Pvr homolog, Mph, that does not function as a poliovirus receptor.
       Previous work has shown that the first immunoglobulin-like domain of the
       Pvr protein contains the virus binding site. To further identify
       sequences of Pvr important for its interaction with poliovirus, stable
       cell lines expressing mutated Pvr molecules were examined for their
       abilities to bind virus and support virus replication. Substitution of
       the amino-terminal domain of Mph with that of Pvr yields a molecule that
       can function as a poliovirus receptor. Cells expressing this chimeric
       receptor have normal binding affinity for poliovirus, yet the kinetics
       of virus replication are delayed. Results of virus alteration assays
       indicate that this chimeric receptor is defective in converting native
       virus to 135S altered particles. This defect is not observed with cells
       expressing receptor recombinants that include Pvr domains 1 and 2.
       Because altered particles are believed to be an intermediate in
       poliovirus entry, these findings suggest that Pvr domains 2 and 3
       participate in early stages of infection. Additional mutants were made
       by substituting variant Mph residues for the corresponding residues in
       Pvr. The results were interpreted by using a model of Pvr predicted from
       the known structures of other immunoglobulin-like V-type domains.
       Analysis of stable cell lines expressing the mutant proteins revealed
       that virus binding is influenced by mutations in the predicted C'-C
       loop, the C beta-strand, the C-D loop, and the D-E loop. Mutations in
       homologous regions of the immunoglobulin-like CD4 molecule alter its
       interaction with gp120 of human immunodeficiency virus type 1. Cells
       expressing Pvr mutations on the predicted C edge do not develop
       cytopathic effect during poliovirus infection, suggesting that
       poliovirus-induced cytopathic effect may be induced by the
       virus-receptor interaction.
 DE    Amino Acid Sequence  Animal  Chimeric Proteins/METABOLISM  Comparative
       Study  Cytopathogenic Effect, Viral/GENETICS  DNA Mutational Analysis
       Flow Cytometry  Hela Cells  Human  L Cells  Mice  Models, Molecular
       Molecular Sequence Data  Mutagenesis, Site-Directed
       Polioviruses/*GROWTH & DEVELOPMENT  Protein Binding  Receptors,
       Virus/ANALYSIS/GENETICS/*METABOLISM  Species Specificity
       Structure-Activity Relationship  Support, Non-U.S. Gov't  Support, U.S.
       Gov't, Non-P.H.S.  Support, U.S. Gov't, P.H.S.  Virus Replication
       JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

