+ 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

The 3'-terminal 55 nucleotides of bovine coronavirus defective interfering RNA harbor cis-acting elements required for both negative- and positive-strand RNA synthesis



The 3'-terminal 55 nucleotides of bovine coronavirus defective interfering RNA harbor cis-acting elements required for both negative- and positive-strand RNA synthesis



Plos one 9(5): E98422



The synthesis of the negative-strand [(-)-strand] complement of the ∼30 kilobase, positive-strand [(+)-strand] coronaviral genome is a necessary early step for genome replication. The identification of cis-acting elements required for (-)-strand RNA synthesis in coronaviruses, however, has been hampered due to insufficiencies in the techniques used to detect the (-)-strand RNA species. Here, we employed a method of head-to-tail ligation and real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) to detect and quantitate the synthesis of bovine coronavirus (BCoV) defective interfering (DI) RNA (-) strands. Furthermore, using the aforementioned techniques along with Northern blot assay, we specifically defined the cis-acting RNA elements within the 3'-terminal 55 nucleotides (nts) which function in the synthesis of (-)- or (+)-strand BCoV DI RNA. The major findings are as follows: (i) nts from -5 to -39 within the 3'-terminal 55 nts are the cis-acting elements responsible for (-)-strand BCoV DI RNA synthesis, (ii) nts from -3 to -34 within the 3'-terminal 55 nts are cis-acting elements required for (+)-strand BCoV DI RNA synthesis, and (iii) the nucleotide species at the 3'-most position (-1) is important, but not critical, for both (-)- and (+)-strand BCoV DI RNA synthesis. These results demonstrate that the 3'-terminal 55 nts in BCoV DI RNA harbor cis-acting RNA elements required for both (-)- and (+)-strand DI RNA synthesis and extend our knowledge on the mechanisms of coronavirus replication. The method of head-to-tail ligation and qRT-PCR employed in the study may also be applied to identify other cis-acting elements required for (-)-strand RNA synthesis in coronaviruses.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 056160176

Download citation: RISBibTeXText

PMID: 24852421


Related references

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

The 5'-terminal region of the Aichi virus genome encodes cis-acting replication elements required for positive- and negative-strand RNA synthesis. Journal of Virology 79(11): 6918-6931, 2005

Importance of the positive-strand RNA secondary structure of a murine coronavirus defective interfering RNA internal replication signal in positive-strand RNA synthesis. Journal of Virology 72(10): 7926-7933, 1998

Enhanced accumulation of coronavirus defective interfering RNA from expressed negative-strand transcripts by coexpressed positive-strand RNA transcripts. Virology 287(2): 286-300, 2001

3'-Terminal sequence in poliovirus negative-strand templates is the primary cis-acting element required for VPgpUpU-primed positive-strand initiation. Journal of Virology 79(6): 3565-3577, 2005

A cis-acting protein is not required for the replication of a coronavirus defective-interfering RNA. Virology 209: 8-36, 1995

Two cis-acting elements in negative RNA strand of Hepatitis C virus involved in synthesis of positive RNA strand in vitro. Acta Virologica 49(2): 83-90, 2005

A cis-acting viral protein is not required for the replication of a coronavirus defective-interfering RNA. Virology 209(2): 428-436, 1995

Replication of murine coronavirus defective interfering RNA from negative-strand transcripts. Journal of Virology 70(9): 5769-5776, 1996

Stem-loop IV in the 5' untranslated region is a cis-acting element in bovine coronavirus defective interfering RNA replication. Journal of Virology 79(19): 12434-12446, 2005

Stem-loop III in the 5' untranslated region is a cis-acting element in bovine coronavirus defective interfering RNA replication. Journal of Virology 77(12): 6720-6730, 2003

An RNA stem-loop within the bovine coronavirus nsp1 coding region is a cis-acting element in defective interfering RNA replication. Journal of Virology 81(14): 7716-7724, 2007

The UCUAAAC promoter motif is not required for high-frequency leader recombination in bovine coronavirus defective interfering RNA. Journal of Virology 70(5): 2720-2729, 1996

Cis-Acting core RNA elements required for negative-strand RNA synthesis and cap-independent translation are separated in the 3'-untranslated region of Red clover necrotic mosaic virus RNA1. Virology 369(1): 168-181, 2007

Poliovirus CRE-dependent VPg uridylylation is required for positive-strand RNA synthesis but not for negative-strand RNA synthesis. Journal of Virology 77(8): 4739-4750, 2003