PMID- 29301233
OWN - NLM
STAT- MEDLINE
DCOM- 20181211
LR  - 20210924
IS  - 1999-4915 (Electronic)
IS  - 1999-4915 (Linking)
VI  - 10
IP  - 1
DP  - 2017 Dec 31
TI  - Analyses of Tissue Culture Adaptation of Human Herpesvirus-6A by Whole Genome 
      Deep Sequencing Redefines the Reference Sequence and Identifies Virus Entry 
      Complex Changes.
LID - 10.3390/v10010016 [doi]
LID - 16
AB  - 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.
FAU - Tweedy, Joshua G
AU  - Tweedy JG
AD  - Department of Pathogen Molecular Biology, London School of Hygiene & Tropical 
      Medicine, University of London, London WC1E 7HT, UK. 
      joshua.tweedy@cruk.manchester.ac.uk.
FAU - Escriva, Eric
AU  - Escriva E
AD  - Department of Pathogen Molecular Biology, London School of Hygiene & Tropical 
      Medicine, University of London, London WC1E 7HT, UK. eric.escriva@lshtm.ac.uk.
AD  - Institute for Structural and Molecular Biology, Department Biology, Birkbeck 
      College University of London, London WC1E 7HX, UK. eric.escriva@lshtm.ac.uk.
FAU - Topf, Maya
AU  - Topf M
AD  - Institute for Structural and Molecular Biology, Department Biology, Birkbeck 
      College University of London, London WC1E 7HX, UK. m.topf@cryst.bbk.ac.uk.
FAU - Gompels, Ursula A
AU  - Gompels UA
AD  - Department of Pathogen Molecular Biology, London School of Hygiene & Tropical 
      Medicine, University of London, London WC1E 7HT, UK. ursula.gompels@lshtm.ac.uk.
LA  - eng
GR  - MR/M019292/1/MRC_/Medical Research Council/United Kingdom
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20171231
PL  - Switzerland
TA  - Viruses
JT  - Viruses
JID - 101509722
RN  - 0 (Viral Envelope Proteins)
SB  - IM
MH  - Adaptation, Physiological/*genetics
MH  - Cell Line
MH  - Genome, Viral/genetics
MH  - Herpesvirus 6, Human/*genetics/physiology
MH  - *High-Throughput Nucleotide Sequencing
MH  - Humans
MH  - Models, Molecular
MH  - Polymorphism, Single Nucleotide
MH  - Roseolovirus Infections/genetics/*virology
MH  - Selection, Genetic
MH  - Sequence Deletion
MH  - Viral Envelope Proteins/chemistry/*genetics
MH  - *Virus Internalization
MH  - *Whole Genome Sequencing
PMC - PMC5795429
OTO - NOTNLM
OT  - HHV-6
OT  - HHV-6A
OT  - Roseolovirus
OT  - betaherpesvirus
OT  - cell fusion
OT  - deep sequencing
OT  - gB
OT  - gH/gL
OT  - glycoprotein structure model
OT  - human herpesvirus
OT  - reference genome
OT  - virus entry complex
COIS- The authors declare no conflict of interest.
EDAT- 2018/01/06 06:00
MHDA- 2018/12/12 06:00
PMCR- 2018/01/01
CRDT- 2018/01/06 06:00
PHST- 2017/11/30 00:00 [received]
PHST- 2017/12/23 00:00 [revised]
PHST- 2017/12/26 00:00 [accepted]
PHST- 2018/01/06 06:00 [entrez]
PHST- 2018/01/06 06:00 [pubmed]
PHST- 2018/12/12 06:00 [medline]
PHST- 2018/01/01 00:00 [pmc-release]
AID - v10010016 [pii]
AID - viruses-10-00016 [pii]
AID - 10.3390/v10010016 [doi]
PST - epublish
SO  - Viruses. 2017 Dec 31;10(1):16. doi: 10.3390/v10010016.