PMID- 20032178
OWN - NLM
STAT- MEDLINE
DCOM- 20100317
LR  - 20211020
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Print)
IS  - 0022-538X (Linking)
VI  - 84
IP  - 5
DP  - 2010 Mar
TI  - Vaccinia virus particles mix inefficiently, and in a way that would restrict 
      viral recombination, in coinfected cells.
PG  - 2432-43
LID - 10.1128/JVI.01998-09 [doi]
AB  - It is well established that poxviruses are subjected to genetic recombination, 
      but attempts to map vaccinia virus genes using classical genetic crosses were 
      historically confounded by high levels of experimental noise and a poor 
      correlation between physical and genetic map distances. These virus-by-virus 
      crosses also never produced the 50% recombinant progeny that should be seen in 
      experiments involving distant markers. Poxviruses replicate in membrane-wrapped 
      cytoplasmic structures called virosomes (or factories) and we have developed a 
      method for tracking the development of these structures using live cell imaging 
      and cells expressing phage lambda Cro protein fused to enhanced green fluorescent 
      protein (EGFP). The EGFP-cro protein binds nonspecifically to DNA and permits 
      live cell imaging of developing vaccinia virus factories. Using this method, we 
      see virosomes first appearing about 4 to 5 h postinfection. The early virosomes 
      exhibit a compact appearance and then, after a period of exponential growth 
      lasting several hours, blur and start to dissipate in a process presumably linked 
      to viral packaging. During the growth period, the virosomes migrate toward the 
      nuclear periphery while colliding and fusing at a rate dependent upon the numbers 
      of infecting particles. However, even at high multiplicities of infection (10 
      PFU/cell), we estimate approximately 20% of the virosomes never fuse. We have 
      also used fluorescence in situ hybridization (FISH) methods to study virosomes 
      formed by the fusion of viruses carrying different gene markers. FISH showed that 
      DNA mixes rather poorly within fused virosomes and the amount of mixing is 
      inversely dependent on the time between virosome appearance and fusion. Our 
      studies suggest that the intracellular movement and mixing of virosomes create 
      constraints that reduce opportunities for forming recombinants and that these 
      phenomena create outcomes reflected in classical poxvirus genetics.
FAU - Lin, Y-C James
AU  - Lin YC
AD  - Alberta Institute for Viral Immunology and Department of Medical Microbiology and 
      Immunology, Faculty of Medicine and Dentistry, The University of Alberta, 
      Alberta, Canada.
FAU - Evans, D H
AU  - Evans DH
LA  - eng
GR  - Canadian Institutes of Health Research/Canada
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20091223
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (DNA, Viral)
RN  - 0 (Genetic Markers)
RN  - 0 (Recombinant Fusion Proteins)
SB  - IM
MH  - DNA, Viral/metabolism
MH  - Genetic Markers
MH  - Genome, Viral
MH  - In Situ Hybridization, Fluorescence
MH  - Recombinant Fusion Proteins/genetics/metabolism
MH  - *Recombination, Genetic
MH  - *Vaccinia virus/genetics/metabolism
MH  - Virion/genetics/*metabolism/ultrastructure
MH  - Virus Assembly
MH  - Virus Replication/physiology
PMC - PMC2820930
EDAT- 2009/12/25 06:00
MHDA- 2010/03/18 06:00
PMCR- 2010/09/01
CRDT- 2009/12/25 06:00
PHST- 2009/12/25 06:00 [entrez]
PHST- 2009/12/25 06:00 [pubmed]
PHST- 2010/03/18 06:00 [medline]
PHST- 2010/09/01 00:00 [pmc-release]
AID - JVI.01998-09 [pii]
AID - 1998-09 [pii]
AID - 10.1128/JVI.01998-09 [doi]
PST - ppublish
SO  - J Virol. 2010 Mar;84(5):2432-43. doi: 10.1128/JVI.01998-09. Epub 2009 Dec 23.