DOI: 10.1128/jvi.74.16.7422-7430.2000. Use of a Transient Assay for Studying the Genetic Determinants of <i>Fv1</i> Restriction

Use of a Transient Assay for Studying the Genetic Determinants of Fv1 Restriction

10.1128/jvi.74.16.7422-7430.2000

Crossref journal-article

American Society for Microbiology

Journal of Virology (235)

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.

Bibliography

Bock, M., Bishop, K. N., Towers, G., & Stoye, J. P. (2000). Use of a Transient Assay for Studying the Genetic Determinants of Fv1 Restriction. Journal of Virology, 74(16), 7422â7430.

Authors 4

Michael Bock (first)
Kate N. Bishop (additional)
Greg Towers (additional)
Jonathan P. Stoye (additional)

References 23 Referenced 123

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Lilly F. Fv-2: identification and location of a second gene governing the spleen focus response to Friend leukemia virus in mice.J. Natl. Cancer Inst. 45 1970 163 169 / J. Natl. Cancer Inst. / Fv-2: identification and location of a second gene governing the spleen focus response to Friend leukemia virus in mice by Lilly F. (1970)
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10.1128/jvi.66.10.5959-5966.1992
Rosenberg N. Jolicoeur P. Retroviral pathogenesis Retroviruses. Coffin J. M. Hughes S. H. Varmus H. E. 1997 475 585 Cold Spring Harbor Laboratory Press Cold Spring Harbor N.Y
Sambrook J. Fritsch E. F. Maniatis T. Molecular cloning: a laboratory manual 2nd ed. 1989 Cold Spring Harbor Laboratory Press Cold Spring Harbor N.Y
10.1093/nar/23.4.628

Dates

Type	When
Created	23 years, 1 month ago (July 27, 2002, 6:06 a.m.)
Deposited	3 years, 5 months ago (March 4, 2022, 11:55 p.m.)
Indexed	4 months, 3 weeks ago (April 9, 2025, 9:31 a.m.)
Issued	25 years ago (Aug. 15, 2000)
Published	25 years ago (Aug. 15, 2000)
Published Print	25 years ago (Aug. 15, 2000)

Funders 0

None

BibTeX

@article{Bock_2000, title={Use of a Transient Assay for Studying the Genetic Determinants of Fv1 Restriction}, volume={74}, ISSN={1098-5514}, url={http://dx.doi.org/10.1128/jvi.74.16.7422-7430.2000}, DOI={10.1128/jvi.74.16.7422-7430.2000}, number={16}, journal={Journal of Virology}, publisher={American Society for Microbiology}, author={Bock, Michael and Bishop, Kate N. and Towers, Greg and Stoye, Jonathan P.}, year={2000}, month=aug, pages={7422–7430} }

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