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Journal of virology1995; 69(5); 2881-2888; doi: 10.1128/JVI.69.5.2881-2888.1995

Replication in vitro and in vivo of an equine infectious anemia virus mutant deficient in dUTPase activity.

Abstract: As an important enzyme in DNA synthesis, dUTPase is present in a wide variety of organisms and viruses and has been identified as a component of the equine infectious anemia virus (EIAV) pol gene. The role of EIAV dUTPase, designated DU, in virus replication in vitro and in vivo was investigated with a recently described infectious molecular clone of EIAV. A deletion mutant that was deficient in dUTPase activity was constructed, and its replication kinetics was examined in fetal equine kidney (FEK) cells and primary equine bone marrow macrophage (EBMM) cells. In FEK cells, which are permissive for EIAV replication, the mutant virus replicated as well as the parental virus. In primary cultures of EBMM cells, which are primary targets of EIAV infection in vivo, the DU mutant showed delayed replication kinetics and replicated to a lower extent than did the parental virus. As the multiplicity of infection decreased, the difference between the parental and mutant viruses increased, such that at the lowest multiplicity of infection tested, there was over a 100-fold difference in virus production. The mutant virus was also much less cytopathic. The role of DU in replication in vivo was examined using a Shetland pony model of EIAV infection. Shetland ponies that were infected with the parental and mutant viruses showed transient virus RNA levels in plasma approximately 5 to 10 days postinfection. The peak virus levels in plasma (as measured by a quantitative reverse transcriptase PCR assay) were 10- to 100-fold lower in the mutant virus-infected animals than in the animals infected with the parental virus. However, ponies infected with the mutant virus mounted similar antibody responses despite the marked differences in virus replication. These studies demonstrate that EIAV DU is important for the efficient replication of the virus in macrophages in vitro and in vivo and suggests that variations in the DU sequence could markedly affect the biological and pathogenic properties of EIAV.
Publication Date: 1995-05-01 PubMed ID: 7707512PubMed Central: PMC188985DOI: 10.1128/JVI.69.5.2881-2888.1995Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • P.H.S.

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research paper investigates the role of the enzyme dUTPase in the replication process of the equine infectious anemia virus (EIAV), using a specially constructed deletion mutant deficient in dUTPase activity. They conducted the research both in vitro and in vivo, using fetal equine kidney (FEK) cells, primary equine bone marrow macrophage (EBMM) cells and Shetland ponies. The EIAV dUTPase, also known as DU, was found to be essential to efficient virus replication, especially in macrophages.

Role and Importance of dUTPase

  • dUTPase is a crucial enzyme in DNA synthesis commonly found in a vast array of organisms and viruses, including EIAV.
  • It is a component of the EIAV pol gene. The study focused on scrutinizing the role of this enzyme, DU, as it pertains to virus replication both in vitro (lab-based) and in vivo (live organisms).

Research Methodology

  • An EIAV mutant lacking dUTPase activity was created for the purpose of this research.
  • The replication process of this mutant virus was then observed in FEK cells and EBMM cells.
  • FEK cells are known for their permissiveness towards EIAV replication. The mutant virus showed similar replication rates to the original EIAV in these cells.
  • EBMM cells, which are the primary targets of EIAV infection in living organisms, showed a different pattern. The lack of DU resulted in delayed replication kinetics and a lower extent of replication compared to the original virus, with the differences growing as the multiplicity of infection decreased.

In Vivo Examination

  • The in vivo aspect of the research used a Shetland pony model of EIAV infection.
  • The plasma of ponies infected with both the parental and mutant viruses showed transient virus RNA levels approximately 5 to 10 days after infection.
  • However, the peak virus levels in the Shetland ponies infected with the mutant virus were significantly lower (10- to 100-fold) than their counterparts infected with the original virus.
  • Interestingly, both groups of ponies exhibited similar antibody responses, despite the marked differences in virus replication.

Conclusion

  • This research provides insight into the crucial role the EIAV DU plays for efficient virus replication, particularly in macrophages, both in vitro and in vivo.
  • The findings suggest that any variations in the DU sequence could significantly influence the biological and pathogenic characteristics of EIAV.

Cite This Article

APA
Lichtenstein DL, Rushlow KE, Cook RF, Raabe ML, Swardson CJ, Kociba GJ, Issel CJ, Montelaro RC. (1995). Replication in vitro and in vivo of an equine infectious anemia virus mutant deficient in dUTPase activity. J Virol, 69(5), 2881-2888. https://doi.org/10.1128/JVI.69.5.2881-2888.1995

Publication

ISSN: 0022-538X
NlmUniqueID: 0113724
Country: United States
Language: English
Volume: 69
Issue: 5
Pages: 2881-2888

Researcher Affiliations

Lichtenstein, D L
  • Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pennsylvania 15261, USA.
Rushlow, K E
    Cook, R F
      Raabe, M L
        Swardson, C J
          Kociba, G J
            Issel, C J
              Montelaro, R C

                MeSH Terms

                • Amino Acid Sequence
                • Animals
                • Antibodies, Viral / biosynthesis
                • Base Sequence
                • Cell Line
                • Cytopathogenic Effect, Viral / genetics
                • Cytopathogenic Effect, Viral / physiology
                • DNA Primers / genetics
                • DNA, Viral / genetics
                • Equine Infectious Anemia / virology
                • Gene Deletion
                • Genes, pol
                • Horses
                • Infectious Anemia Virus, Equine / enzymology
                • Infectious Anemia Virus, Equine / genetics
                • Infectious Anemia Virus, Equine / physiology
                • Molecular Sequence Data
                • Polymerase Chain Reaction
                • Pyrophosphatases / deficiency
                • Pyrophosphatases / genetics
                • RNA, Viral / blood
                • RNA, Viral / genetics
                • Virus Replication / genetics
                • Virus Replication / physiology

                Grant Funding

                • AI 25850 / NIAID NIH HHS
                • CA 49296 / NCI NIH HHS

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