PMID- 27548280
OWN - NLM
STAT- MEDLINE
DCOM- 20170623
LR  - 20190202
IS  - 1553-7374 (Electronic)
IS  - 1553-7366 (Print)
IS  - 1553-7366 (Linking)
VI  - 12
IP  - 8
DP  - 2016 Aug
TI  - Single-Molecule FISH Reveals Non-selective Packaging of Rift Valley Fever Virus 
      Genome Segments.
PG  - e1005800
LID - 10.1371/journal.ppat.1005800 [doi]
LID - e1005800
AB  - The bunyavirus genome comprises a small (S), medium (M), and large (L) RNA 
      segment of negative polarity. Although genome segmentation confers evolutionary 
      advantages by enabling genome reassortment events with related viruses, genome 
      segmentation also complicates genome replication and packaging. Accumulating 
      evidence suggests that genomes of viruses with eight or more genome segments are 
      incorporated into virions by highly selective processes. Remarkably, little is 
      known about the genome packaging process of the tri-segmented bunyaviruses. Here, 
      we evaluated, by single-molecule RNA fluorescence in situ hybridization (FISH), 
      the intracellular spatio-temporal distribution and replication kinetics of the 
      Rift Valley fever virus (RVFV) genome and determined the segment composition of 
      mature virions. The results reveal that the RVFV genome segments start to 
      replicate near the site of infection before spreading and replicating throughout 
      the cytoplasm followed by translocation to the virion assembly site at the Golgi 
      network. Despite the average intracellular S, M and L genome segments approached 
      a 1:1:1 ratio, major differences in genome segment ratios were observed among 
      cells. We also observed a significant amount of cells lacking evidence of 
      M-segment replication. Analysis of two-segmented replicons and four-segmented 
      viruses subsequently confirmed the previous notion that Golgi recruitment is 
      mediated by the Gn glycoprotein. The absence of colocalization of the different 
      segments in the cytoplasm and the successful rescue of a tri-segmented variant 
      with a codon shuffled M-segment suggested that inter-segment interactions are 
      unlikely to drive the copackaging of the different segments into a single virion. 
      The latter was confirmed by direct visualization of RNPs inside mature virions 
      which showed that the majority of virions lack one or more genome segments. 
      Altogether, this study suggests that RVFV genome packaging is a non-selective 
      process.
FAU - Wichgers Schreur, Paul J
AU  - Wichgers Schreur PJ
AD  - Department of Virology, Central Veterinary Institute, part of Wageningen 
      University and Research Centre, Lelystad, The Netherlands.
FAU - Kortekaas, Jeroen
AU  - Kortekaas J
AD  - Department of Virology, Central Veterinary Institute, part of Wageningen 
      University and Research Centre, Lelystad, The Netherlands.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20160822
PL  - United States
TA  - PLoS Pathog
JT  - PLoS pathogens
JID - 101238921
SB  - IM
MH  - Cell Line
MH  - Genome, Viral/*physiology
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Rift Valley Fever/metabolism
MH  - Rift Valley fever virus/*physiology
MH  - Virus Assembly/*physiology
PMC - PMC4993503
COIS- The authors have declared that no competing interests exist.
EDAT- 2016/08/23 06:00
MHDA- 2017/06/24 06:00
PMCR- 2016/08/22
CRDT- 2016/08/23 06:00
PHST- 2016/02/26 00:00 [received]
PHST- 2016/07/09 00:00 [accepted]
PHST- 2016/08/23 06:00 [entrez]
PHST- 2016/08/23 06:00 [pubmed]
PHST- 2017/06/24 06:00 [medline]
PHST- 2016/08/22 00:00 [pmc-release]
AID - PPATHOGENS-D-16-00462 [pii]
AID - 10.1371/journal.ppat.1005800 [doi]
PST - epublish
SO  - PLoS Pathog. 2016 Aug 22;12(8):e1005800. doi: 10.1371/journal.ppat.1005800. 
      eCollection 2016 Aug.