Abstract
Cytopathic feline leukemia virus (FeLV) infections of feline T-cell line (FeT-cell) cultures led to the accumulation and maintenance of threefold more proviruses with deletions within the polymerase gene (pol) than minimally cytopathic FeLV infections. Over 60% of the viral DNA from cytopathic infections bore deletions in pol. Characterization of DNA sequences adjoining the deletions revealed that the junctions were most often flanked by RNA splice donor and acceptor consensus motifs. A thymidine-to-cytidine mutation introduced at the +2 position of one RNA splice donor-like motif inhibited formation of the two most prevalent viral DNA species with deletions, confirming the origin of many proviruses with deletions from reverse transcription of aberrantly spliced viral RNA species. An example of deletion by misalignment was also characterized. Viral inocula obtained from cells recovered after cytopathic infections were attenuated in their ability to cause cytopathic effects (CPE) and were able to confer superinfection resistance to naïve FeT-cells, despite maintaining envelope gene (env) sequences with full cytopathic potential. This suggested that viral genomes with deletions, rather than being required for cytopathicity, play a role in protecting cells from CPE. Indeed, expression of a molecularly cloned provirus bearing one of the characterized deletions attenuated CPE in FeT-cells caused by superinfecting cytopathic virus.
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- Ahmed R., Chakraborty P. R., Fields B. N. Genetic variation during lytic reovirus infection: high-passage stocks of wild-type reovirus contain temperature-sensitive mutants. J Virol. 1980 Apr;34(1):285–287. doi: 10.1128/jvi.34.1.285-287.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Aziz D. C., Hanna Z., Jolicoeur P. Severe immunodeficiency disease induced by a defective murine leukaemia virus. Nature. 1989 Apr 6;338(6215):505–508. doi: 10.1038/338505a0. [DOI] [PubMed] [Google Scholar]
- Berberich S. L., Stoltzfus C. M. Analysis of spliced and unspliced Rous sarcoma virus RNAs early and late after infection of chicken embryo fibroblasts: effect of cell culture conditions. Virology. 1991 May;182(1):135–144. doi: 10.1016/0042-6822(91)90657-w. [DOI] [PubMed] [Google Scholar]
- Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ciminale V., Pavlakis G. N., Derse D., Cunningham C. P., Felber B. K. Complex splicing in the human T-cell leukemia virus (HTLV) family of retroviruses: novel mRNAs and proteins produced by HTLV type I. J Virol. 1992 Mar;66(3):1737–1745. doi: 10.1128/jvi.66.3.1737-1745.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Coffin J. M., Tsichlis P. N., Barker C. S., Voynow S., Robinson H. L. Variation in avian retrovirus genomes. Ann N Y Acad Sci. 1980;354:410–425. doi: 10.1111/j.1749-6632.1980.tb27982.x. [DOI] [PubMed] [Google Scholar]
- Crandell R. A., Fabricant C. G., Nelson-Rees W. A. Development, characterization, and viral susceptibility of a feline (Felis catus) renal cell line (CRFK). In Vitro. 1973 Nov-Dec;9(3):176–185. doi: 10.1007/BF02618435. [DOI] [PubMed] [Google Scholar]
- Doggett D. L., Drake A. L., Hirsch V., Rowe M. E., Stallard V., Mullins J. I. Structure, origin, and transforming activity of feline leukemia virus-myc recombinant provirus FTT. J Virol. 1989 May;63(5):2108–2117. doi: 10.1128/jvi.63.5.2108-2117.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Donahue P. R., Hoover E. A., Beltz G. A., Riedel N., Hirsch V. M., Overbaugh J., Mullins J. I. Strong sequence conservation among horizontally transmissible, minimally pathogenic feline leukemia viruses. J Virol. 1988 Mar;62(3):722–731. doi: 10.1128/jvi.62.3.722-731.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Donahue P. R., Quackenbush S. L., Gallo M. V., deNoronha C. M., Overbaugh J., Hoover E. A., Mullins J. I. Viral genetic determinants of T-cell killing and immunodeficiency disease induction by the feline leukemia virus FeLV-FAIDS. J Virol. 1991 Aug;65(8):4461–4469. doi: 10.1128/jvi.65.8.4461-4469.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fischer H. D., Dodgson J. B., Hughes S., Engel J. D. An unusual 5' splice sequence is efficiently utilized in vivo. Proc Natl Acad Sci U S A. 1984 May;81(9):2733–2737. doi: 10.1073/pnas.81.9.2733. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hartman S. C., Mulligan R. C. Two dominant-acting selectable markers for gene transfer studies in mammalian cells. Proc Natl Acad Sci U S A. 1988 Nov;85(21):8047–8051. doi: 10.1073/pnas.85.21.8047. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Henderson I. C., Lieber M. M., Todaro G. J. Mink cell line Mv 1 Lu (CCL 64). Focus formation and the generation of "nonproducer" transformed cell lines with murine and feline sarcoma viruses. Virology. 1974 Jul;60(1):282–287. doi: 10.1016/0042-6822(74)90386-9. [DOI] [PubMed] [Google Scholar]
- Hu W. S., Temin H. M. Retroviral recombination and reverse transcription. Science. 1990 Nov 30;250(4985):1227–1233. doi: 10.1126/science.1700865. [DOI] [PubMed] [Google Scholar]
- Jackson I. J. A reappraisal of non-consensus mRNA splice sites. Nucleic Acids Res. 1991 Jul 25;19(14):3795–3798. doi: 10.1093/nar/19.14.3795. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Karacostas V., Wolffe E. J., Nagashima K., Gonda M. A., Moss B. Overexpression of the HIV-1 gag-pol polyprotein results in intracellular activation of HIV-1 protease and inhibition of assembly and budding of virus-like particles. Virology. 1993 Apr;193(2):661–671. doi: 10.1006/viro.1993.1174. [DOI] [PubMed] [Google Scholar]
- Katz R. A., Kotler M., Skalka A. M. cis-acting intron mutations that affect the efficiency of avian retroviral RNA splicing: implication for mechanisms of control. J Virol. 1988 Aug;62(8):2686–2695. doi: 10.1128/jvi.62.8.2686-2695.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kristal B. S., Reinhart T. A., Hoover E. A., Mullins J. I. Interference with superinfection and with cell killing and determination of host range and growth kinetics mediated by feline leukemia virus surface glycoproteins. J Virol. 1993 Jul;67(7):4142–4153. doi: 10.1128/jvi.67.7.4142-4153.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mount S. M. A catalogue of splice junction sequences. Nucleic Acids Res. 1982 Jan 22;10(2):459–472. doi: 10.1093/nar/10.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mullins J. I., Chen C. S., Hoover E. A. Disease-specific and tissue-specific production of unintegrated feline leukaemia virus variant DNA in feline AIDS. Nature. 1986 Jan 23;319(6051):333–336. doi: 10.1038/319333a0. [DOI] [PubMed] [Google Scholar]
- Overbaugh J., Donahue P. R., Quackenbush S. L., Hoover E. A., Mullins J. I. Molecular cloning of a feline leukemia virus that induces fatal immunodeficiency disease in cats. Science. 1988 Feb 19;239(4842):906–910. doi: 10.1126/science.2893454. [DOI] [PubMed] [Google Scholar]
- Overbaugh J., Hoover E. A., Mullins J. I., Burns D. P., Rudensey L., Quackenbush S. L., Stallard V., Donahue P. R. Structure and pathogenicity of individual variants within an immunodeficiency disease-inducing isolate of FeLV. Virology. 1992 Jun;188(2):558–569. doi: 10.1016/0042-6822(92)90510-v. [DOI] [PubMed] [Google Scholar]
- Papenhausen M. D., Overbaugh J. Nucleotide sequence of the splice junction of feline leukemia virus envelope mRNA. Virology. 1993 Aug;195(2):804–807. doi: 10.1006/viro.1993.1434. [DOI] [PubMed] [Google Scholar]
- Pulsinelli G. A., Temin H. M. Characterization of large deletions occurring during a single round of retrovirus vector replication: novel deletion mechanism involving errors in strand transfer. J Virol. 1991 Sep;65(9):4786–4797. doi: 10.1128/jvi.65.9.4786-4797.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reinhart T. A., Ghosh A. K., Hoover E. A., Mullins J. I. Distinct superinfection interference properties yet similar receptor utilization by cytopathic and noncytopathic feline leukemia viruses. J Virol. 1993 Sep;67(9):5153–5162. doi: 10.1128/jvi.67.9.5153-5162.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanger F., Coulson A. R. A rapid method for determining sequences in DNA by primed synthesis with DNA polymerase. J Mol Biol. 1975 May 25;94(3):441–448. doi: 10.1016/0022-2836(75)90213-2. [DOI] [PubMed] [Google Scholar]
- Schwartz S., Felber B. K., Benko D. M., Fenyö E. M., Pavlakis G. N. Cloning and functional analysis of multiply spliced mRNA species of human immunodeficiency virus type 1. J Virol. 1990 Jun;64(6):2519–2529. doi: 10.1128/jvi.64.6.2519-2529.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shimotohno K., Temin H. M. Loss of intervening sequences in genomic mouse alpha-globin DNA inserted in an infectious retrovirus vector. Nature. 1982 Sep 16;299(5880):265–268. doi: 10.1038/299265a0. [DOI] [PubMed] [Google Scholar]
- Sitbon M., d'Auriol L., Ellerbrok H., André C., Nishio J., Perryman S., Pozo F., Hayes S. F., Wehrly K., Tambourin P. Substitution of leucine for isoleucine in a sequence highly conserved among retroviral envelope surface glycoproteins attenuates the lytic effect of the Friend murine leukemia virus. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5932–5936. doi: 10.1073/pnas.88.13.5932. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Snyder H. W., Jr, Hardy W. D., Jr, Zuckerman E. E., Fleissner E. Characterisation of a tumour-specific antigen on the surface of feline lymphosarcoma cells. Nature. 1978 Oct 19;275(5681):656–658. doi: 10.1038/275656a0. [DOI] [PubMed] [Google Scholar]
- Sorge J., Hughes S. H. Splicing of intervening sequences introduced into an infectious retroviral vector. J Mol Appl Genet. 1982;1(6):547–559. [PubMed] [Google Scholar]
- Spindler K. R., Horodyski F. M., Holland J. J. High multiplicities of infection favor rapid and random evolution of vesicular stomatitis virus. Virology. 1982 May;119(1):96–108. doi: 10.1016/0042-6822(82)90068-x. [DOI] [PubMed] [Google Scholar]
- VON MAGNUS P. Incomplete forms of influenza virus. Adv Virus Res. 1954;2:59–79. doi: 10.1016/s0065-3527(08)60529-1. [DOI] [PubMed] [Google Scholar]
- Viglianti G. A., Mullins J. I. Functional comparison of transactivation by simian immunodeficiency virus from rhesus macaques and human immunodeficiency virus type 1. J Virol. 1988 Dec;62(12):4523–4532. doi: 10.1128/jvi.62.12.4523-4532.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Viglianti G. A., Sharma P. L., Mullins J. I. Simian immunodeficiency virus displays complex patterns of RNA splicing. J Virol. 1990 Sep;64(9):4207–4216. doi: 10.1128/jvi.64.9.4207-4216.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weiss R. A. AIDS. Defective viruses to blame? Nature. 1989 Apr 6;338(6215):458–458. doi: 10.1038/338458a0. [DOI] [PubMed] [Google Scholar]
- Youngner J. S., Jones E. V., Kelly M., Frielle D. W. Generation and amplification of temperature-sensitive mutants during serial undiluted passages of vesicular stomatitis virus. Virology. 1981 Jan 15;108(1):87–97. doi: 10.1016/0042-6822(81)90529-8. [DOI] [PubMed] [Google Scholar]
- de Noronha C. M., Mullins J. I. Amplimers with 3'-terminal phosphorothioate linkages resist degradation by vent polymerase and reduce Taq polymerase mispriming. PCR Methods Appl. 1992 Nov;2(2):131–136. doi: 10.1101/gr.2.2.131. [DOI] [PubMed] [Google Scholar]
