+ 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

Requirement of the 5'-end genomic sequence as an upstream cis-acting element for coronavirus subgenomic mRNA transcription

Requirement of the 5'-end genomic sequence as an upstream cis-acting element for coronavirus subgenomic mRNA transcription

Journal of Virology 68(8): 4727-4737

We have developed a defective interfering (DI) RNA containing a chloramphenicol acetyltransferase reporter gene, placed behind an intergenic sequence, for studying subgenomic mRNA transcription of mouse hepatitis virus (MHV), a prototype coronavirus. Using this system, we have identified the sequence requirement for MHV subgenomic mRNA transcription. We show that this sequence requirement differs from that for RNA replication. In addition to the previously identified requirement for an intergenic (promoter) sequence, additional sequences from the 5' end of genomic RNA are required for subgenomic mRNA transcription. These upstream sequences include the leader RNA and a spacer sequence between the leader and intergenic sequence, which is derived from the 5' untranslated region and part of gene 1. The spacer sequence requirement is specific, since only the sequence derived from the 5' end of RNA genome, but not from other MHV genomic regions or heterologous sequences, could initiate subgenomic transcription from the intergenic sequence. These results strongly suggest that the wild-type viral subgenomic mRNAs (mRNA2 to mRNA7) and probably their counterpart subgenomic negative-sense RNAs cannot be utilized for mRNA amplification. Furthermore, we have demonstrated that a partial leader sequence present at the 5' end of genome, which lacks the leader-mRNA fusion sequence, could still support subgenomic mRNA transcription. In this case, the leader sequences of the subgenomic transcripts were derived exclusively from the wild-type helper virus, indicating that the MHV leader RNA initiates in trans subgenomic mRNA transcription. Thus, the leader sequence can enhance subgenomic transcription even when it cannot serve as a primer for mRNA synthesis. These results taken together suggest that the 5'-end leader sequence of MHV not only provides a trans-acting primer for mRNA initiation but also serves as a cis-acting element required for the transcription of subgenomic mRNAs. The identification of an upstream cis-acting element for MHV subgenomic mRNA synthesis defines a novel sequence requirement for regulating mRNA synthesis in RNA viruses.

Please choose payment method:

(PDF emailed within 1 workday: $29.90)

Accession: 009342960

Download citation: RISBibTeXText

PMID: 8035475

Related references

The leader RNA of coronavirus mouse hepatitis virus contains an enhancer-like element for subgenomic mRNA transcription. Journal of Virology 74(22): 10571-10580, 2000

Role of nucleotides immediately flanking the transcription-regulating sequence core in coronavirus subgenomic mRNA synthesis. Journal of Virology 79(4): 2506-2516, 2005

Coronavirus mRNA transcription: UV light transcriptional mapping studies suggest an early requirement for a genomic-length template. Journal of Virology 66(8): 4671-4678, 1992

Identification of a noncanonical signal for transcription of a novel subgenomic mRNA of mouse hepatitis virus: implication for the mechanism of coronavirus RNA transcription. Virology 278(1): 75-85, 2000

A 5'-proximal RNA sequence of murine coronavirus as a potential initiation site for genomic-length mRNA transcription. Journal of Virology 70(2): 705-711, 1996

Identification of an upstream sequence element required for vesicular stomatitis virus mRNA transcription. Journal of Virology 76(15): 7632-7641, 2002

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

Coronavirus leader RNA regulates and initiates subgenomic mRNA transcription both in trans and in cis. Journal of Virology 68(8): 4738-4746, 1994

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

The 3' untranslated region of coronavirus RNA is required for subgenomic mRNA transcription from a defective interfering RNA. Journal of Virology 70(10): 7236-7240, 1996

Upstream induction sequence the cis acting element required for response to the allantoin pathway inducer and enhancement of operation of the nitrogen regulated upstream activation sequence in saccharomyces cerevisiae. Journal of Bacteriology 173(22): 7186-7195, 1991

Investigation of the control of coronavirus subgenomic mRNA transcription by using T7-generated negative-sense RNA transcripts. Journal of Virology 69(10): 6219-6227, 1995

Downstream sequences influence the choice between a naturally occurring noncanonical and closely positioned upstream canonical heptameric fusion motif during bovine coronavirus subgenomic mRNA synthesis. Journal of Virology 75(16): 7362-7374, 2001

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

Mutual interference between genomic RNA replication and subgenomic mRNA transcription in brome mosaic virus. Journal of Virology 79(3): 1438-1451, 2005