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    Analyses of tissue culture adaptation of human Herpesvirus-6A by whole genome deep sequencing redefines the reference sequence and identifies virus entry complex changes

    Tweedy, J. and Escriva, E. and Topf, Maya and Gompels, U. (2017) Analyses of tissue culture adaptation of human Herpesvirus-6A by whole genome deep sequencing redefines the reference sequence and identifies virus entry complex changes. Viruses 10 (1), p. 16. ISSN 1999-4915.

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    Abstract

    Tissue-culture adaptation of viruses can modulate infection. Laboratory passage and bacterial artificial chromosome (BAC)mid cloning of human cytomegalovirus, HCMV, resulted in genomic deletions and rearrangements altering genes encoding the virus entry complex, which affected cellular tropism, virulence, and vaccine development. Here, we analyse these effects on the reference genome for related betaherpesviruses, Roseolovirus, human herpesvirus 6A (HHV-6A) strain U1102. This virus is also naturally “cloned” by germline subtelomeric chromosomal-integration in approximately 1% of human populations, and accurate references are key to understanding pathological relationships between exogenous and endogenous virus. Using whole genome next-generation deep-sequencing Illumina-based methods, we compared the original isolate to tissue-culture passaged and the BACmid-cloned virus. This re-defined the reference genome showing 32 corrections and 5 polymorphisms. Furthermore, minor variant analyses of passaged and BACmid virus identified emerging populations of a further 32 single nucleotide polymorphisms (SNPs) in 10 loci, half non-synonymous indicating cell-culture selection. Analyses of the BAC-virus genome showed deletion of the BAC cassette via loxP recombination removing green fluorescent protein (GFP)-based selection. As shown for HCMV culture effects, select HHV-6A SNPs mapped to genes encoding mediators of virus cellular entry, including virus envelope glycoprotein genes gB and the gH/gL complex. Comparative models suggest stabilisation of the post-fusion conformation. These SNPs are essential to consider in vaccine-design, antimicrobial-resistance, and pathogenesis.

    Metadata

    Item Type: Article
    Additional Information: This article belongs to the Special Issue Breakthroughs in Viral Replication
    Keyword(s) / Subject(s): human herpesvirus, HHV-6, HHV-6A, Roseolovirus, betaherpesvirus, virus entry complex, gH/gL, gB, deep sequencing, reference genome, cell fusion, glycoprotein structure model
    School: Birkbeck Schools and Departments > School of Science > Biological Sciences
    Depositing User: Administrator
    Date Deposited: 25 Jan 2018 15:22
    Last Modified: 25 Jan 2018 15:22
    URI: http://eprints.bbk.ac.uk/id/eprint/21026

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