+ 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

Characterization of a murine coronavirus defective interfering RNA internal cis-acting replication signal



Characterization of a murine coronavirus defective interfering RNA internal cis-acting replication signal



Journal of Virology 69(8): 4963-4971



The mouse hepatitis virus (MHV) sequences required for replication of the JHM strain of MHV defective interfering (DI) RNA consist of three discontinuous genomic regions: about 0.47 kb from both terminal sequences and a 0.13-kb internal region present at about 0.9 kb from the 5' end of the DI genome. In this study, we investigated the role of the internal 0.13-kb region in MHV RNA replication. Overall sequences of the 0.13-kb regions from various MHV strains were similar to each other, with nucleotide substitutions in some strains; MHV-A59 was exceptional, with three nucleotide deletions. Computer-based secondary-structure analysis of the 0.13-kb region in the positive strand revealed that most of the MHV strains formed the same or a similar main stem-loop structure, whereas only MHV-A59 formed a smaller main stem-loop structure. The RNA secondary structures in the negative strands were much less uniform among the MHV strains. A series of DI RNAs that contained MHV-JHM-derived 5'- and 3'-terminal sequences plus internal 0.13-kb regions derived from various MHV strains were constructed. Most of these DI RNAs replicated in MHV-infected cells, except that MRP-A59, with a 0.13-kb region derived from MHV-A59, failed to replicate. Interestingly, replication of MRP-A59 was temperature dependent; it occurred at 39.5 degrees C but not at 37 or 35 degrees C, whereas a DI RNA with an MHV-JHM-derived 0.13-kb region replicated at all three temperatures. At 37 degrees C, synthesis of MRP-A59 negative-strand RNA was detected in MHV-infected and MRP-A59 RNA-transfected cells. Another DI RNA with the internal 0.13-kb region deleted also synthesized negative-strand RNA in MHV-infected cells. MRP-A59-transfected cells were shifted from 39.5 to 37 degrees C at 5.5 h postinfection, a time when most MHV negative-strand RNAs have already accumulated; after the shift, MRP-A59 positive-strand RNA synthesis ceased. The minimum sequence required for maintenance of the positive-strand major stem-loop structure and biological function of the MHV-JHM 0.13-kb region was about 57 nucleotides. Function was lost in the 50-nucleotide sequence that formed a positive-strand stem-loop structure identical to that of MHV-A59. These studies suggested that the RNA structure made by the internal sequence was important for positive-strand MHV RNA synthesis.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 008294554

Download citation: RISBibTeXText

PMID: 7609066


Related references

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

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

A cis-acting function for the coronavirus leader in defective interfering RNA replication. Journal of Virology 68(12): 8223-8231, 1994

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

Analysis of cis-acting sequences essential for coronavirus defective interfering RNA replication. Virology 197(1): 53-63, 1993

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

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

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

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

Analysis of efficiently packaged defective interfering RNAs of murine coronavirus: localization of a possible RNA-packaging signal. Journal of Virology 64(12): 6045-6053, 1990

Defective-interfering particles of murine coronavirus: mechanism of synthesis of defective viral RNAs. Virology 163(1): 104-111, 1988

Replication of vesicular stomatitis virus defective interfering particle rna in vitro transition from synthesis of defective interfering leader rna to synthesis of full length defective interfering rna. Journal of Virology 46(2): 513-522, 1983

The 3' cis-acting genomic replication element of the severe acute respiratory syndrome coronavirus can function in the murine coronavirus genome. Journal of Virology 78(14): 7846-7851, 2004