The polypyrimidine tract-binding protein affects coronavirus RNA accumulation levels and relocalizes viral RNAs to novel cytoplasmic domains different from replication-transcription sites

Sola, Isabel, Galan, Carmen, Mateos-Gomez, Pedro A., Palacio, Lorena, Zuniga, Sonia, Cruz, Jazmina L., Almazan, Fernando and Enjuanes, Luis (2011) The polypyrimidine tract-binding protein affects coronavirus RNA accumulation levels and relocalizes viral RNAs to novel cytoplasmic domains different from replication-transcription sites. Journal of Virology, 85 10: 5136-5149. doi:10.1128/JVI.00195-11

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Author Sola, Isabel
Galan, Carmen
Mateos-Gomez, Pedro A.
Palacio, Lorena
Zuniga, Sonia
Cruz, Jazmina L.
Almazan, Fernando
Enjuanes, Luis
Title The polypyrimidine tract-binding protein affects coronavirus RNA accumulation levels and relocalizes viral RNAs to novel cytoplasmic domains different from replication-transcription sites
Journal name Journal of Virology   Check publisher's open access policy
ISSN 0022-538X
Publication date 2011
Sub-type Article (original research)
DOI 10.1128/JVI.00195-11
Open Access Status File (Publisher version)
Volume 85
Issue 10
Start page 5136
End page 5149
Total pages 14
Place of publication Washington, DC United States
Publisher American Society for Microbiology
Abstract The coronavirus (CoV) discontinuous transcription mechanism is driven by long-distance RNA-RNA interactions between transcription-regulating sequences (TRSs) located at the 5' terminal leader (TRS-L) and also preceding each mRNA-coding sequence (TRS-B). The contribution of host cell proteins to CoV transcription needs additional information. Polypyrimidine tract-binding protein (PTB) was reproducibly identified in association with positive-sense RNAs of transmissible gastroenteritis coronavirus (TGEV) TRS-L and TRS-B by affinity chromatography and mass spectrometry. A temporal regulation of PTB cytoplasmic levels was observed during infection, with a significant increase from 7 to 16 h postinfection being inversely associated with a decrease in viral replication and transcription. Silencing the expression of PTB with small interfering RNA in two cell lines (Huh7 and HEK 293T) led to a significant increase of up to 4-fold in mRNA levels and virus titer, indicating a negative effect of PTB on CoV RNA accumulation. During CoV infection, PTB relocalized from the nucleus to novel cytoplasmic structures different from replication-transcription sites in which stress granule markers T-cell intracellular antigen-1 (TIA-1) and TIA-1-related protein (TIAR) colo-calized. PTB was detected in these modified stress granules in TGEV-infected swine testis cells but not in stress granules induced by oxidative stress. Furthermore, viral genomic and subgenomic RNAs were detected in association with PTB and TIAR. These cytoplasmic ribonucleoprotein complexes might be involved in post-transcriptional regulation of virus gene expression.
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: UQ Diamantina Institute Publications
 
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