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Increased neurovirulence and reactivation of the herpes simplex virus type 1 latency-associated transcript (LAT)-negative mutant dLAT2903 with a disrupted LAT miR-H2

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Abstract

At least six microRNAs (miRNAs) appear to be encoded by the latency-associated transcript (LAT) of herpes simplex virus type 1 (HSV-1). The gene for ICP0, an important immediate early (IE) viral protein, is anti-sense to, and overlaps with, the region of LAT from which miRNA H2 (miR-H2) is derived. We recently reported that a mutant (McK-ΔH2) disrupted for miR-H2 on the wild-type HSV-1 strain McKrae genomic background has increased ICP0 expression, increased neurovirulence, and slightly more rapid reactivation. We report here that HSV-1 mutants deleted for the LAT promoter nonetheless make significant amounts of miR-H2 during lytic tissue culture infection, presumably via readthrough transcription from an upstream promoter. To determine if miR-H2 might also play a role in the HSV-1 latency/reactivation cycle of a LAT-negative mutant, we constructed dLAT-ΔH2, in which miR-H2 is disrupted in dLAT2903 without altering the predicted amino acid sequence of the overlapping ICP0 open reading frame. Similar to McK-ΔH2, dLAT-ΔH2 expressed more ICP0, was more neurovirulent, and had increased reactivation in the mouse TG explant-induced reactivation model of HSV-1 compared with its parental virus. Interestingly, although the increased reactivation of McK-ΔH2 compared with its parental wild-type (wt) virus was subtle and only detected at very early times after explant TG induced reactivation, the increased reactivation of dLAT-ΔH2 compared with its dLAT2903 parental virus appeared more robust and was significantly increased even at late times after induction. These results confirm that miR-H2 plays a role in modulating the HSV-1 reactivation phenotype.

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Acknowledgments

This study was supported by Public Health Service NIH grants 1R56AI098985, 1R56AI093133, R01EY013191, RO1EY019896, RO1EY14900, and EY024618 and The Discovery Center for Eye Research. We thank Dr. Nigel Fraser for reading this manuscript and providing helpful comments.

Conflict of interest

All of the authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Virology Research, Gavin Herbert Eye Institute and Department of Ophthalmology, University of California Irvine, School of Medicine, Irvine, CA, 92697, USA

    Xianzhi Jiang, Don Brown, Nelson Osorio, Chinhui Hsiang, Lbachir BenMohamed & Steven L. Wechsler

  2. Laboratory of Cellular and Molecular Immunology, Gavin Herbert Eye Institute and Department of Ophthalmology, University of California Irvine, School of Medicine, Irvine, CA, 92697, USA

    Lbachir BenMohamed & Steven L. Wechsler

  3. Chao Family Comprehensive Cancer Center, University of California Irvine Medical Center, Irvine, CA, 92868-32013, USA

    Lbachir BenMohamed

  4. Institute for Immunology, University of California Irvine, School of Medicine, Irvine, CA, 92697, USA

    Lbachir BenMohamed

  5. Department of Microbiology and Molecular Genetics, University of California Irvine, School of Medicine, Irvine, CA, 92697, USA

    Steven L. Wechsler

  6. Center for Virus Research, University of California Irvine, Irvine, CA, 92697, USA

    Steven L. Wechsler

  7. Ophthalmology Research, University of California Irvine, 843 Health Sciences Road, Hewitt Hall (Building 843), Room 2012, Irvine, CA, 92697, USA

    Steven L. Wechsler

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  1. Xianzhi Jiang
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  2. Don Brown
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  3. Nelson Osorio
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  4. Chinhui Hsiang
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Correspondence to Steven L. Wechsler.

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Jiang, X., Brown, D., Osorio, N. et al. Increased neurovirulence and reactivation of the herpes simplex virus type 1 latency-associated transcript (LAT)-negative mutant dLAT2903 with a disrupted LAT miR-H2. J. Neurovirol. 22, 38–49 (2016). https://doi.org/10.1007/s13365-015-0362-y

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