July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
The absense of DHHC3 affects primary and latent HSV-1 infection
Author Affiliations & Notes
  • Shaohui Wang
    Cedars Sinai Medical Center, West Hollywood, California, United States
  • Kevin Mott
    Cedars Sinai Medical Center, West Hollywood, California, United States
  • Marianne Cilluffo
    UCLA, Los Angeles, California, United States
  • Casey Kilpatrick
    Pennsylvania State University, University Park, Pennsylvania, United States
  • Shoko Murakami
    Pennsylvania State University, University Park, Pennsylvania, United States
  • Alexander V Ljubimov
    Cedars Sinai Medical Center, West Hollywood, California, United States
  • Konstantin G. Kousoulas
    Louisiana State University, Baton Rouge, Louisiana, United States
  • Sita Awasthi
    University of Pennsylvania, Philadelphia, Pennsylvania, United States
  • Bernhard Luscher
    Pennsylvania State University, University Park, Pennsylvania, United States
  • Homayon Ghiasi
    Cedars Sinai Medical Center, West Hollywood, California, United States
  • Footnotes
    Commercial Relationships   Shaohui Wang, None; Kevin Mott, None; Marianne Cilluffo, None; Casey Kilpatrick, None; Shoko Murakami, None; Alexander Ljubimov, None; Konstantin Kousoulas, None; Sita Awasthi, None; Bernhard Luscher, None; Homayon Ghiasi, None
  • Footnotes
    Support  This work was supported by Public Health Service grant RO1 EY13615 from the National Eye Institute.
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 514. doi:https://doi.org/
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      Shaohui Wang, Kevin Mott, Marianne Cilluffo, Casey Kilpatrick, Shoko Murakami, Alexander V Ljubimov, Konstantin G. Kousoulas, Sita Awasthi, Bernhard Luscher, Homayon Ghiasi; The absense of DHHC3 affects primary and latent HSV-1 infection. Invest. Ophthalmol. Vis. Sci. 2018;59(9):514. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : We reported recently that UL20 can bind to a host protein encoded by the zinc finger DHHC-type containing 3 (ZDHHC3) gene (also known as Golgi-specific DHHC zinc finger protein {GODZ}). The goal for this study is to further investigate the role of GODZ in HSV-1 virus replication and infectivity using GODZ knockout mice model and MEF cells isolated from GODZ knockout mice.

Methods : MEF cells were prepared from GODZ knockout mice. The virus titer in tear film of infected mice were collected by eye swap and virus titer were quantified by standard virus plaque assay. Immunocytochemistry and IHC were used to study virus infectivity and the localization of varies HSV-1 virus proteins within the MEF cells and tissue sections. The virus replication in cornea were analyzed by direct image of GFP positive corneas. The transcripts of virus genes in eyes and TGs of infected mice were quantified by qPCR. The palmitoylation of UL20 in HSV-1 infected WT or GODZ knockout MEF cells was analyzed by palmitoylation assay. The virions in MEF cells were imaged by Electron Microscopy.

Results : The absence of GODZ resulted in reduced palmitoylation of UL20 and altered localization, expression of UL20 and gK; the expression of gB and gC; and the localization and expression of tegument and capsid proteins within HSV-1 infected MEFs. Electron microscopy revealed that the absence of GODZ limited the maturation of virions at multiple steps, and affected the localization of virus and endoplasmic reticulum morphology. Virus replication in the eyes of ocularly HSV-1 infected GODZ-/- mice was significantly lower than in HSV-1 WT mice. The levels of UL20, gK and gB transcripts in the corneas of HSV-1 infected GODZ-/- mice on day 5 post infection were markedly lower than in WT mice, whereas only UL20 transcripts were reduced in trigeminal ganglia(TG). In addition, HSV-1 infected GODZZ-/- mice showed notably lower levels of corneal scarring.

Conclusions : The absence of GODZ affects primary and latent infection as well as reactivation in ocularly infected mice. The reduced virus infectivity is due to the absence of HSV-1 UL20 binding to GODZ. These results strongly suggest that binding of UL20 to GODZ promotes virus infectivity in vitro and viral pathogenesis in vivo, thus blocking the binding of UL20 to GODZ could be a potential therapeutic target for HSV-1.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

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