Skip to Main Content
 

Global Search Box

 
 
 
 

Files

File List


Thesis.1.pdf (3.59 MB)

ETD Abstract Container

Abstract Header

Impact of VHSV M Protein on the Innate Immune System

Weaver, Wade G.
http://rave.ohiolink.edu/etdc/view?acc_num=toledo1481191320713471

Abstract Details

2016, Master of Science, University of Toledo, Biology (Cell-Molecular Biology).
Viral Hemorrhagic Septicemia Virus (VHSv) is a highly contagious and deadly fish virus that has been found in over 80 different host species. VHSv replicates entirely in the cytoplasm and has a linear, single stranded, negative sense RNA genome that encodes five structural proteins: nucleocapsid protein (N), polymerase associated phosphoprotein (P), matrix protein (M), glycoprotein (G) and virus polymerase protein (L). The sixth gene codes for the nonstructural nonvirion (NV) protein, which is unique to a small group of virus belonging to the genus Novirhabdoviridae. As with all viruses, VHSv must overcome the host innate immune response in order to survive. As such, our data suggest that VHSv can inhibit IFN-stimulated antiviral responses. Inhibition of host transcription is a common activity of many ssRNA viruses, and our previous studies have found that VHSv IVb M also potently inhibited host transcription. In order to further study VHSv M protein function and mechanism of action, we found that changes at D62 or D62/E181 greatly reduced the anti-transcriptional function of M. With this information and 3D molecular modelling based prediction that these mutations would not negatively impact VHSv M structure, the two mutant M proteins (M_D62A and M_D62/E181A) were incorporated into rWT VHS virus using a reverse genetic system. We have since evaluated these mutant M containing viruses for characteristics of an attenuated virus that could be used in vaccine development. We observed that the M_D62A virus has reduced anti-transcriptional function while retaining WT replicative abilities and allowed the highest total level of antiviral gene activity. Conversely, the M_2x Mut. virus showed reduction in both anti-transcriptional and replicative capacities while allowing for the highest level of antiviral gene activity per virion. These data suggest that D62A is important for anti-host functions of VHSv M while it has little involvement in pro-virus functions, while the opposite is true for residue E181. Taken together, the M_2x Mut. virus could provide a valuable platform from which to launch further studies aimed at generating an attenuated virus vaccine. M_D62A virus should be used in further mutational studies aimed at determining the mechanism of VHSv M transcription inhibition. The studies described in this thesis have improved our understanding of the role that VHSv M has in both inhibiting host transcription, and viral replication.
Douglas Leaman, Ph.D. (Advisor)
Malathi Krishnamurthy, Ph.D. (Committee Member)
Heather Conti, Ph.D. (Committee Member)
Travis Taylor, Ph.D. (Committee Member)
92 p.
Biology
VHSv, Matrix Protein, Transcription

Recommended Citations

Citations

  • Weaver, W. G. (2016). Impact of VHSV M Protein on the Innate Immune System [Master's thesis, University of Toledo]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=toledo1481191320713471

    APA Style (7th edition)

  • Weaver, Wade. Impact of VHSV M Protein on the Innate Immune System. 2016. University of Toledo, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=toledo1481191320713471.

    MLA Style (8th edition)

  • Weaver, Wade. "Impact of VHSV M Protein on the Innate Immune System." Master's thesis, University of Toledo, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=toledo1481191320713471

    Chicago Manual of Style (17th edition)

toledo1481191320713471
325
© 2016, all rights reserved.
This open access ETD is published by University of Toledo and OhioLINK.