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Characterization of the S1, S2, S3 and DU open reading frames of equine infectious anemia virus

Steagall, Wendy Kay
http://rave.ohiolink.edu/etdc/view?acc_num=case1061991340

Abstract Details

1995, Doctor of Philosophy, Case Western Reserve University, Molecular Biology and Microbiology.
Equine Infectious Anemia Virus (EIAV) contains three small open reading frames, designated S1, S2, and S3, that may encode regulatory proteins. To determine the functions of these open reading frames, cDNA clones were isolated and characterized. Three classes of cDNA clones were obtained. The first class contains the leader region, S1, and S3, while the second class contains the leader region, S1, part of env, and S3. The leader-S1 reading frame encodes Tat activity, while the env-S3 region encodes Rev activity. The third class of cDNA contains part of the gag gene fused to part of the env gene, and its function is undetermined. No cDNA clone was isolated that primarily encodes S2, and the function of S2 is unknown. EIAV also encodes a dUTPase from the DU domain present in the pol gene. The inability of EIAV containing a 270 bp deletion in the DU domain (Δ DU) to replicate to wild type (WT) levels in primary equine macrophages has been described (Threadgill et al., 1993). In this dissertation, I describe the construction of a second dUTPase-deficient virus containing a single amino acid substitution in dUTPase (DUD71E). This virus showed a similar growth phenotype to Δ DU in macrophages, replicating to only 2% of WT levels. T he blocks to viral replication of Δ DU and DUD71E in macrophages were also examined. Reverse transcription occurred normally in macrophages with full length viral DNA evident by 72 hours post-infection; however, the Δ DU EIAV incorporated uracil into the viral DNA, as shown by in vitro and in vivo assays, while WT EIAV did not. The viral DNA from the dUTPase-deficient viruses integrated into macrophage DNA, but at levels 2- to 3-fold less than WT levels. Steady state levels of viral transcripts were severely decreased in macrophages infected with dUTPase-deficient viruses as compared to WT EIAV. These results suggest that the major block to dUTPase-deficient viral replication in macrophages occurs between integration and viral transcription. A virally encoded dUTPase may be necessary to prevent incorporation of uracil into viral DNA in nondividing macrophages, which may have low levels of this enzyme
Susan Payne (Advisor)
140 p.
Biology, Molecular
Infectious anemia virus, equine; Reading frames

Recommended Citations

Citations

  • Steagall, W. K. (1995). Characterization of the S1, S2, S3 and DU open reading frames of equine infectious anemia virus [Doctoral dissertation, Case Western Reserve University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=case1061991340

    APA Style (7th edition)

  • Steagall, Wendy. Characterization of the S1, S2, S3 and DU open reading frames of equine infectious anemia virus. 1995. Case Western Reserve University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=case1061991340.

    MLA Style (8th edition)

  • Steagall, Wendy. "Characterization of the S1, S2, S3 and DU open reading frames of equine infectious anemia virus." Doctoral dissertation, Case Western Reserve University, 1995. http://rave.ohiolink.edu/etdc/view?acc_num=case1061991340

    Chicago Manual of Style (17th edition)

case1061991340
435
© 1995, all rights reserved.
This open access ETD is published by Case Western Reserve University School of Graduate Studies and OhioLINK.