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Pathogenesis and Immunity

Marek’s Disease Virus Requires Both Copies of the Inverted Repeat Regions for Efficient In Vivo Replication and Pathogenesis

Tereza Vychodil, Andelé M. Conradie, Jakob Trimpert, Amr Aswad, Luca D. Bertzbach, Benedikt B. Kaufer
Felicia Goodrum, Editor
Tereza Vychodil
aInstitut für Virologie, Freie Universität Berlin, Berlin, Germany
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Andelé M. Conradie
aInstitut für Virologie, Freie Universität Berlin, Berlin, Germany
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Jakob Trimpert
aInstitut für Virologie, Freie Universität Berlin, Berlin, Germany
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Amr Aswad
aInstitut für Virologie, Freie Universität Berlin, Berlin, Germany
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Luca D. Bertzbach
aInstitut für Virologie, Freie Universität Berlin, Berlin, Germany
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  • ORCID record for Luca D. Bertzbach
Benedikt B. Kaufer
aInstitut für Virologie, Freie Universität Berlin, Berlin, Germany
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Felicia Goodrum
University of Arizona
Roles: Editor
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DOI: 10.1128/JVI.01256-20
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ABSTRACT

Marek’s disease virus (MDV) is an oncogenic alphaherpesvirus of chickens. The MDV genome consists of two unique regions that are both flanked by inverted repeat regions. These repeats harbor several genes involved in virus replication and pathogenesis, but it remains unclear why MDV and other herpesviruses harbor these large sequence duplications. In this study, we set to determine if both copies of these repeat regions are required for MDV replication and pathogenesis. Our results demonstrate that MDV mutants lacking the entire internal repeat region (ΔIRLS) efficiently replicate and spread from cell-to-cell in vitro. However, ΔIRLS replication was severely impaired in infected chickens and the virus caused significantly less frequent disease and tumors compared to the controls. In addition, we also generated recombinant viruses that harbor a deletion of most of the internal repeat region, leaving only short terminal sequences behind (ΔIRLS-HR). These remaining homologous sequences facilitated rapid restoration of the deleted repeat region, resulting in a virus that caused disease and tumors comparable to the wild type. Therefore, ΔIRLS-HR represents an excellent platform for rapid genetic manipulation of the virus genome in the repeat regions. Taken together, our study demonstrates that MDV requires both copies of the repeats for efficient replication and pathogenesis in its natural host.

IMPORTANCE Marek’s disease virus (MDV) is a highly oncogenic alphaherpesvirus that infects chickens and causes losses in the poultry industry of up to $2 billion per year. The virus is also widely used as a model to study alphaherpesvirus pathogenesis and virus-induced tumor development in a natural host. MDV and most other herpesviruses harbor direct or inverted repeats regions in their genome. However, the role of these sequence duplications in MDV remains elusive and has never been investigated in a natural virus-host model for any herpesvirus. Here, we demonstrate that both copies of the repeats are needed for efficient MDV replication and pathogenesis in vivo, while replication was not affected in cell culture. With this, we further dissect herpesvirus genome biology and the role of repeat regions in Marek’s disease virus replication and pathogenesis.

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Marek’s Disease Virus Requires Both Copies of the Inverted Repeat Regions for Efficient In Vivo Replication and Pathogenesis
Tereza Vychodil, Andelé M. Conradie, Jakob Trimpert, Amr Aswad, Luca D. Bertzbach, Benedikt B. Kaufer
Journal of Virology Jan 2021, 95 (3) e01256-20; DOI: 10.1128/JVI.01256-20

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Marek’s Disease Virus Requires Both Copies of the Inverted Repeat Regions for Efficient In Vivo Replication and Pathogenesis
Tereza Vychodil, Andelé M. Conradie, Jakob Trimpert, Amr Aswad, Luca D. Bertzbach, Benedikt B. Kaufer
Journal of Virology Jan 2021, 95 (3) e01256-20; DOI: 10.1128/JVI.01256-20
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KEYWORDS

Marek’s disease virus
alphaherpesvirus
repeat regions
diploid genes
replication
class E genome
recombination
genome restoration
cancer
inverted repeat regions
pathogenesis

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