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Home / Equine Research Bank / J Virol / Characterization of equine infectious anemia virus dUTPase: ...
Journal of virology1993; 67(5); 2592-2600; doi: 10.1128/JVI.67.5.2592-2600.1993

Characterization of equine infectious anemia virus dUTPase: growth properties of a dUTPase-deficient mutant.

Abstract: The putative dUTPase domain was deleted from the polymerase (pol) gene of equine infectious anemia virus (EIAV) to produce a recombinant delta DUpol Escherichia coli expression cassette and a delta DU proviral clone. Expression of the recombinant delta DUpol polyprotein yielded a properly processed and enzymatically active reverse transcriptase, as determined by immunoblot analysis and DNA polymerase activity gels. Transfection of delta DU provirus into feline (FEA) cells resulted in production of virus that replicated to wild-type levels in both FEA cells and fetal equine kidney cells. In contrast, the delta DU virus replicated poorly (less than 1% of wild-type levels) in primary equine macrophage cultures, as measured by reverse transcriptase assays. Preparations of delta DU virus contained negligible dUTPase activity, which confirms that virion-associated dUTPase is encoded in the pol gene region between the RNase H domain and integrase, as has been demonstrated previously for feline immunodeficiency virus (J. H. Elder, D. L. Lerner, C. S. Hasselkus-Light, D. J. Fontenot, E. Hunter, P. A. Luciw, R. C. Montelaro, and T. R. Phillips, J. Virol. 66:1791-1794, 1992). Our results suggest that virus-encoded dUTPase is dispensable for virus replication in dividing cells in vitro but may be required for efficient replication of EIAV in nondividing equine macrophages, the natural host cells for this virus.
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Publication Date: 1993-05-01 PubMed ID: 8386267PubMed Central: PMC237580DOI: 10.1128/JVI.67.5.2592-2600.1993Google Scholar: Lookup
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  • Comparative Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • Non-P.H.S.
  • 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.

The researchers studied the role of dUTPase, a potential protein domain, within the equine infectious anemia virus (EIAV). By deleting this domain, they found that the altered virus replicates effectively in certain cell types but not others, suggesting this component may be integral for the virus’s effective reproduction in its natural host cells.

Research Methodology

  • The study involved the deletion of the putative dUTPase domain from the polymerase (pol) gene of EIAV, leading to the production of a recombinant delta DUpol Escherichia coli expression cassette and a delta DU proviral clone.
  • The expression of this altered version of the polyprotein was tested for enzymatic activity and processing accuracy using immunoblot analysis and DNA polymerase activity gels.
  • The modified provirus was transfected into feline (FEA) cells for the observation of the viral behaviour after the removal of the dUTPase domain.

Findings

  • The results showed that the mutant virus could replicate to wild-type levels in FEA cells and fetal equine kidney cells.
  • In stark contrast, the delta DU virus displayed poor replication (less than 1% of wild-type levels) in primary equine macrophage cultures which was measured through reverse transcriptase assays.
  • The study confirmed that dUTPase activity was dramatically reduced in the mutant virus. This finding demonstrated that the dUTPase domain is coded within the pol gene region between the RNase H domain and integrase, aligning with past research findings on the feline immunodeficiency virus.

Implications

  • This study suggests that the virus-encoded dUTPase plays a significant role in virus replication especially in non-dividing equine macrophages, the natural host cells for EIAV.
  • Therefore, it may be of potential interest to focus on this protein domain in approaches aimed at controlling the infection and treatment of the equine infectious anemia virus.

Cite This Article

APA
Threadgill DS, Steagall WK, Flaherty MT, Fuller FJ, Perry ST, Rushlow KE, Le Grice SF, Payne SL. (1993). Characterization of equine infectious anemia virus dUTPase: growth properties of a dUTPase-deficient mutant. J Virol, 67(5), 2592-2600. https://doi.org/10.1128/JVI.67.5.2592-2600.1993

Publication

Journal of virology
ISSN: 0022-538X
NlmUniqueID: 0113724
Country: United States
Language: English
Volume: 67
Issue: 5
Pages: 2592-2600

Researcher Affiliations

Threadgill, D S
  • Department of Molecular Biology and Microbiology, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106-4960.
Steagall, W K
    Flaherty, M T
      Fuller, F J
        Perry, S T
          Rushlow, K E
            Le Grice, S F
              Payne, S L

                MeSH Terms

                • Amino Acid Sequence
                • Animals
                • Base Sequence
                • Cats
                • Cytopathogenic Effect, Viral
                • Escherichia coli / genetics
                • Gene Deletion
                • Gene Products, pol / biosynthesis
                • Horses
                • Infectious Anemia Virus, Equine / enzymology
                • Infectious Anemia Virus, Equine / genetics
                • Infectious Anemia Virus, Equine / growth & development
                • Molecular Sequence Data
                • Protein Processing, Post-Translational
                • Pyrophosphatases / deficiency
                • Pyrophosphatases / genetics
                • Recombinant Fusion Proteins / biosynthesis
                • Virus Replication

                Grant Funding

                • AI07381-03 / NIAID NIH HHS
                • AI31147 / NIAID NIH HHS
                • CA50168-03 / NCI NIH HHS

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