Detection of proviral DNA in horse cells infected with equine infectious anemia virus.
Abstract: Equine infectious anemia virus (EIAV) recently has been shown to possess a high-molecular-weight RNA genome and a virion reverse transcriptase. We completed the demonstration that EIAV is a retrovirus by showing the presence of proviral DNA in equine cells infected in vitro, but not in normal horse DNA. These studies were performed by using a highly representative cDNA probe synthesized by the virion polymerase. It was found that this cDNA reassociated extensively, and with high thermal stability, with either viral RNA or DNA extracted from infected cells, but showed no detectable reassociation with DNA from uninfected horse cells. Similarly, sequences related to EIAV were neither found in the DNA of four other Equus species, nor in a variety of other mammals including sheep, cows, pigs- dogs, cats, and humans; nor did EIAV cDNA hybridize with a variety of other retrovirus RNAs. These experiments were performed under conditions of very low stringency to enable detection of distantly related sequences, with a sufficient ratio of DAN to cDNA to allow detection of less than one viral copy per haploid genome. We conclude that EIAV is not an endogenous virus of the horse or of the other species tested.
Publication Date: 1978-06-01 PubMed ID: 209211PubMed Central: PMC525883DOI: 10.1128/JVI.26.3.577-583.1978Google Scholar: Lookup
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- Journal Article
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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The research demonstrates that the Equine Infectious Anemia Virus (EIAV) possesses proviral DNA in infected horse cells, but not in uninfected equine DNA, confirming it as a type of retrovirus. The study used a cDNA probe related to EIAV in several animals and found no significant reassociation, suggesting that EIAV is not an endogenous virus in these species.
Identification of Proviral DNA in EIAV
- The researchers conducted this study by infecting equine cells in vitro with EIAV. The goal was to confirm the retroviral properties of EIAV, which include the presence of high-molecular-weight RNA genome and a virion reverse transcriptase.
- The results were affirmative as proviral DNA was found in infected equine cells. However, such proviral DNA was not found in healthy horse DNA, providing the first evidence of EIAV’s retroviral nature.
Use of cDNA Probe
- The researchers utilized a cDNA probe that was highly representative and synthesized by the virion polymerase to complete the research.
- This cDNA probe was found to have an extensive and thermally stable reassociation with either the viral RNA or DNA derived from infected cells. On the other hand, no detectable reassociation was found with DNA from uninfected horse cells.
Testing Similarity of EIAV with Other Retroviruses
- The researchers then expanded the study to check the presence of EIAV-related sequences in the DNA of four other Equus species, and mammals including sheep, cows, pigs, dogs, cats, and humans. This was done using very low stringency conditions to detect even distantly related sequences. They ensured a sufficient ratio of DNA to cDNA for detecting less than one viral copy per haploid genome.
- However, no EIAV related sequences were found in the tested species’ DNA. Furthermore, the EIAV cDNA probe did not hybridize with RNAs from a variety of retroviruses. This led the researchers to conclude that EIAV is not an endogenous virus of any of these species.
Cite This Article
APA
Rice NR, Simek S, Ryder OA, Coggins L.
(1978).
Detection of proviral DNA in horse cells infected with equine infectious anemia virus.
J Virol, 26(3), 577-583.
https://doi.org/10.1128/JVI.26.3.577-583.1978 Publication
Researcher Affiliations
MeSH Terms
- Animals
- Cell Line
- DNA / analysis
- DNA, Viral / analysis
- Horses
- Infectious Anemia Virus, Equine / analysis
- Infectious Anemia Virus, Equine / growth & development
- Nucleic Acid Conformation
- Nucleic Acid Hybridization
- RNA, Viral / analysis
- Species Specificity
References
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Citations
This article has been cited 13 times.- Harrold SM, Cook SJ, Cook RF, Rushlow KE, Issel CJ, Montelaro RC. Tissue sites of persistent infection and active replication of equine infectious anemia virus during acute disease and asymptomatic infection in experimentally infected equids. J Virol 2000 Apr;74(7):3112-21.
- Sellon DC, Fuller FJ, McGuire TC. The immunopathogenesis of equine infectious anemia virus. Virus Res 1994 May;32(2):111-38.
- Tainsky MA. Analysis of the virogenes related to the rhesus monkey endogenous type C retrovirus in monkeys and apes. J Virol 1981 Mar;37(3):922-30.
- Simek S, Rice NR. Analysis of the nucleic acid components in reticuloendotheliosis virus. J Virol 1980 Jan;33(1):320-9.
- Montelaro RC, Lohrey N, Parekh B, Blakeney EW, Issel CJ. Isolation and comparative biochemical properties of the major internal polypeptides of equine infectious anemia virus. J Virol 1982 Jun;42(3):1029-38.
- Salinovich O, Payne SL, Montelaro RC, Hussain KA, Issel CJ, Schnorr KL. Rapid emergence of novel antigenic and genetic variants of equine infectious anemia virus during persistent infection. J Virol 1986 Jan;57(1):71-80.
- Casey JM, Kim Y, Andersen PR, Watson KF, Fox JL, Devare SG. Human T-cell lymphotropic virus type III: immunologic characterization and primary structure analysis of the major internal protein, p24. J Virol 1985 Aug;55(2):417-23.
- Rwambo PM, Issel CJ, Adams WV Jr, Hussain KA, Miller M, Montelaro RC. Equine infectious anemia virus (EIAV) humoral responses of recipient ponies and antigenic variation during persistent infection. Arch Virol 1990;111(3-4):199-212.
- Rasty S, Dhruva BR, Schiltz RL, Shih DS, Issel CJ, Montelaro RC. Proviral DNA integration and transcriptional patterns of equine infectious anemia virus during persistent and cytopathic infections. J Virol 1990 Jan;64(1):86-95.
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- Charman H, Long C, Coggins L. Specificity of response to viral proteins in horses infected with equine infectious anemia virus. Infect Immun 1979 Feb;23(2):472-8.
- Rice NR, Coggins L. Synthesis of long complementary DNA in the endogenous reaction by equine infectious anemia virus. J Virol 1979 Mar;29(3):907-14.
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