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Silencing Suppression by Herpes Simplex Virus Type 1

Wu, Zetang
http://rave.ohiolink.edu/etdc/view?acc_num=osu1213287215

Abstract Details

2008, Doctor of Philosophy, Ohio State University, Molecular, Cellular, and Developmental Biology.
It was hypothesized that herpes simplex virus type-1 (HSV-1) encodes one or more silencing suppressors during productive infection. A transient silencing system was developed that relies on co-transfection of mammalian cells with a plasmid that expresses a silencing target (EGFP) and one that expresses either a perfect EGFP-specific hairpin (dsEGFP) or an imperfect hairpin (midsEGFP). Co-transfection of target plasmid with a plasmid that expresses perfect or imperfect hairpins specific for lacZ (dsLacZ or midsLacZ, respectively), was used as a control. dsEGFP and midsEGFP reduced the EGFP mRNA level by 2.5- and 7-fold, respectively, compared with control hairpins, and a 1:1 molar ratio of midsEGFP to target plasmid yielded maximum silencing of EGFP. Silencing by the midsEGFP hairpin decreased the half-life of EGFP mRNA ~2-fold compared with the control imperfect hairpin. Infection of transfected cells with HSV-1 wild-type strain KOS increased the steady-state amount of EGFP mRNA, regardless of whether or not the EGFP transcript contained an intron. This increase was due, at least in part, to an increase in the half-life of EGFP mRNA in silenced cells, consistent with silencing suppression. Increased accumulation of target EGFP mRNA occurred as early as 4 hr after HSV-1 infection and did not require viral DNA synthesis. The virion component, US11, can suppress silencing in plants in the absence of other HSV-1 genes. Further experiments suggest that US11 could play an important role in silencing suppression in mammalian cells. Taken together these results demonstrate that infection by HSV-1 can suppress established silencing and suggest that this silencing suppression may have evolved as a means to counter a very early host silencing response following virus infection. Indeed, the yields of virus following HSV-1 infection of cells treated with siRNA to Ago-2, compared to those treated with RISC-free control RNA duplex, were significantly increased.
Deborah Parris, PhD (Advisor)
Biao Ding, PhD (Committee Member)
Kathleen Boris-Lawrie, PhD (Committee Member)
David Bisaro, PhD (Committee Member)
151 p.
Virology
HSV-1; RNA silencing; silencing suppression

Recommended Citations

Citations

  • Wu, Z. (2008). Silencing Suppression by Herpes Simplex Virus Type 1 [Doctoral dissertation, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1213287215

    APA Style (7th edition)

  • Wu, Zetang. Silencing Suppression by Herpes Simplex Virus Type 1. 2008. Ohio State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1213287215.

    MLA Style (8th edition)

  • Wu, Zetang. "Silencing Suppression by Herpes Simplex Virus Type 1." Doctoral dissertation, Ohio State University, 2008. http://rave.ohiolink.edu/etdc/view?acc_num=osu1213287215

    Chicago Manual of Style (17th edition)

osu1213287215
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© 2008, all rights reserved.
This open access ETD is published by The Ohio State University and OhioLINK.