Affiliation:
1. Division of Virology, National Institute for Medical Research, London NW7 1AA, United Kingdom
Abstract
ABSTRACT
To probe the genetic determinants controlling the interaction between the retroviral restriction gene
Fv1
and its murine leukemia virus target, we set out to develop rapid, transient assays for
Fv1
function. Cells were transfected or transduced with
Fv1
expression plasmids which can produce green fluorescent protein via an internal ribosome entry site positioned between the
Fv1
and green fluorescent protein coding sequences.
Fv1
function was then assessed by comparing virus replication in green fluorescent protein-positive and -negative cells, using retroviral vectors encoding a second fluorescent marker, yellow fluorescent protein, or β-galactosidase. Using this assay, we could show that
Fv1
specificities were not as absolute as previously thought, since the
Fv1
b
allele was capable of interacting with “nonrestricted” B- and NB-tropic viruses and by shuffling the n- and b-alleles of
Fv1
, it was possible to generate a Fv1 molecule capable of restricting N-, B-, and NB-tropic viruses equally efficiently. Further, we could show that the presence of nonrestricting Fv1 in the same cell as restrictive Fv1 abrogates restriction, implying competition for binding to the retroviral target.
Publisher
American Society for Microbiology
Subject
Virology,Insect Science,Immunology,Microbiology
Cited by
124 articles.
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