+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Coronavirus genomic and subgenomic minus-strand RNAs copartition in membrane-protected replication complexes



Coronavirus genomic and subgenomic minus-strand RNAs copartition in membrane-protected replication complexes



Journal of Virology 71(10): 7744-7749



The majority of porcine transmissible gastroenteritis coronavirus plus-strand RNAs (genome and subgenomic mRNAs), at the time of peak RNA synthesis (5 h postinfection), were not found in membrane-protected complexes in lysates of cells prepared by Dounce homogenization but were found to be susceptible to micrococcal nuclease (85%) or to sediment to a pellet in a cesium chloride gradient (61%). They therefore are probably free molecules in solution or components of easily dissociable complexes. By contrast, the majority of minus-strand RNAs (genome length and subgenomic mRNA length) were found to be resistant to micrococcal nuclease (69%) or to remain suspended in association with membrane-protected complexes following isopycnic sedimentation in a cesium chloride gradient (85%). Furthermore, 35% of the suspended minus strands were in a dense complex (1.20 to 1.24 g/ml) that contained an RNA plus-to-minus-strand molar ratio of approximately 8:1 and viral structural proteins S, M, and N, and 65% were in a light complex (1.15 to 1.17 g/ml) that contained nearly equimolar amounts of plus- and minus-strand RNAs and only trace amounts of proteins M and N. In no instance during fractionation were genome-length minus strands found segregated from sub-genome-length minus strands. These results indicate that all minus-strand species are components of similarly structured membrane-associated replication complexes and support the concept that all are active in the synthesis of plus-strand RNAs.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 008391179

Download citation: RISBibTeXText

PMID: 9311859


Related references

Coronavirus subgenomic minus-strand RNAs and the potential for mRNA replicons. Proceedings of the National Academy of Sciences of the United States of America 86(14): 5626-5630, 1989

Role of subgenomic minus-strand RNA in coronavirus replication. Archives of Virology. Supplementum 9: 173-180, 1994

Identification of the cis-acting signal for minus-strand RNA synthesis of a murine coronavirus: implications for the role of minus-strand RNA in RNA replication and transcription. Journal of Virology 68(12): 8131-8140, 1994

The 5' end of coronavirus minus-strand RNAs contains a short poly(U) tract. Journal of Virology 65(11): 6331-6333, 1991

Importance of coronavirus negative-strand genomic RNA synthesis prior to subgenomic RNA transcription. Virus Research 57(1): 35-42, 1998

SARS-coronavirus replication/transcription complexes are membrane-protected and need a host factor for activity in vitro. Plos Pathogens 4(5): E1000054, 2008

Interferon-gamma inhibits replication of subgenomic and genomic hepatitis C virus RNAs. Hepatology 35(3): 694-703, 2002

Replicase-binding sites on plus- and minus-strand brome mosaic virus RNAs and their roles in RNA replication in plant cells. Journal of Virology 78(24): 13420-9, 2004

Identification of cis-acting elements on positive-strand subgenomic mRNA required for the synthesis of negative-strand counterpart in bovine coronavirus. Viruses 6(8): 2938-2959, 2014

Synthesis of genomic and subgenomic RNAs by a membrane-bound RNA-dependent RNA polymerase isolated from oat plants infected with cereal yellow dwarf virus. Archives of Virology 151(11): 2229-2242, 2006

Fate of minus-strand templates and replication complexes produced by a p23-cleavage-defective mutant of Sindbis virus. Journal of Virology 83(17): 8553-8564, 2009

Altered balance of the synthesis of plus- and minus-strand RNAs induced by RNAs 1 and 2 of alfalfa mosaic virus in the absence of RNA 3. Virology 124(1): 75-85, 1983

Similarities and differences between the subgenomic and minus-strand promoters of an RNA plant virus. Journal of Virology 78(8): 4048-4053, 2004

Polypeptide requirements for assembly of functional Sindbis virus replication complexes: a model for the temporal regulation of minus- and plus-strand RNA synthesis. Embo Journal 13(12): 2925-2934, 1994

Identification and characterization of severe acute respiratory syndrome coronavirus subgenomic RNAs. Advances in Experimental Medicine and Biology 581: 85-88, 2006