PMID- 21715502
OWN - NLM
STAT- MEDLINE
DCOM- 20111004
LR  - 20211020
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Print)
IS  - 0022-538X (Linking)
VI  - 85
IP  - 17
DP  - 2011 Sep
TI  - Mouse hepatitis virus stem-loop 4 functions as a spacer element required to drive 
      subgenomic RNA synthesis.
PG  - 9199-209
LID - 10.1128/JVI.05092-11 [doi]
AB  - The 5' 140 nucleotides of the mouse hepatitis virus (MHV) 5' untranslated region 
      (5'UTR) are predicted to contain three secondary structures, stem-loop 1 (SL1), 
      SL2, and SL4. SL1 and SL2 are required for subgenomic RNA synthesis. The current 
      study focuses on SL4, which contains two base-paired regions, SL4a and SL4b. A 
      series of reverse genetic experiments show that SL4a is not required to be base 
      paired. Neither the structure, the sequence, nor the putative 8-amino-acid open 
      reading frame (ORF) in SL4b is required for viral replication. Viruses containing 
      separate deletions of SL4a and SL4b are viable. However, deletion of SL4 is 
      lethal, and genomes carrying this deletion are defective in directing subgenomic 
      RNA synthesis. Deletion of (131)ACA(133) just 3' to SL4 has a profound impact on 
      viral replication. Viruses carrying the (131)ACA(133) deletion were heterogeneous 
      in plaque size. We isolated three viruses with second-site mutations in the 5'UTR 
      which compensated for decreased plaque sizes, delayed growth kinetics, and lower 
      titers associated with the (131)ACA(133) deletion. The second-site mutations are 
      predicted to change either the spacing between SL1 and SL2 or that between SL2 
      and SL4 or to destabilize the proximal portion of SL4a in our model. A mutant 
      constructed by replacing SL4 with a shorter sequence-unrelated stem-loop was 
      viable. These results suggest that the proposed SL4 in the MHV 5'UTR functions in 
      part as a spacer element that orients SL1, SL2, and the transcriptional 
      regulatory sequence (TRS), and this spacer function may play an important role in 
      directing subgenomic RNA synthesis.
FAU - Yang, Dong
AU  - Yang D
AD  - Department of Microbial and Molecular Pathogenesis, Texas A&M University 
      System-HSC, College of Medicine, College Station, TX 77843-1114, USA.
FAU - Liu, Pinghua
AU  - Liu P
FAU - Giedroc, David P
AU  - Giedroc DP
FAU - Leibowitz, Julian
AU  - Leibowitz J
LA  - eng
GR  - R01 AI051493/AI/NIAID NIH HHS/United States
GR  - R01 AI067416/AI/NIAID NIH HHS/United States
GR  - AI067416/AI/NIAID NIH HHS/United States
GR  - AI051493/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20110629
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (5' Untranslated Regions)
RN  - 0 (RNA, Viral)
SB  - IM
MH  - *5' Untranslated Regions
MH  - Animals
MH  - Base Pairing
MH  - Microbial Viability
MH  - Models, Molecular
MH  - Murine hepatitis virus/genetics/growth & development/*physiology
MH  - Mutation
MH  - Nucleic Acid Conformation
MH  - Open Reading Frames
MH  - RNA, Viral/*genetics/*metabolism
MH  - Sequence Deletion
MH  - Suppression, Genetic
MH  - *Transcription, Genetic
MH  - Viral Plaque Assay
MH  - *Virus Replication
PMC - PMC3165806
EDAT- 2011/07/01 06:00
MHDA- 2011/10/05 06:00
PMCR- 2012/03/01
CRDT- 2011/07/01 06:00
PHST- 2011/07/01 06:00 [entrez]
PHST- 2011/07/01 06:00 [pubmed]
PHST- 2011/10/05 06:00 [medline]
PHST- 2012/03/01 00:00 [pmc-release]
AID - JVI.05092-11 [pii]
AID - 5092-11 [pii]
AID - 10.1128/JVI.05092-11 [doi]
PST - ppublish
SO  - J Virol. 2011 Sep;85(17):9199-209. doi: 10.1128/JVI.05092-11. Epub 2011 Jun 29.