Nascent Synthesis of Leader Sequence-Containing Subgenomic mRNAs in Coronavirus Genome-Length Replicative Intermediate RNA

Mizutani, Tetsuya; Repass, John; Makino, Shinji

doi:10.1006/viro.2000.0489

Cited by 16 publications

(13 citation statements)

References 20 publications

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“…Recently, strong experimental evidence supporting the discontinuous transcription during negative-strand RNA synthesis has been reported [3,62]. Also the leader-primed transcription has received additional support [41].…”

Section: Introductionmentioning

confidence: 99%

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et al. 2001

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The complete sequence (28580 nt) of the PUR46-MAD clone of the Purdue cluster of transmissible gastroenteritis coronavirus (TGEV) has been determined and compared with members of this cluster and other coronaviruses. The computing distances among their S gene sequences resulted in the grouping of these coronaviruses into four clusters, one of them exclusively formed by the Purdue viruses. Three new potential sequence motifs with homology to the alpha-subunit of the polymerase-associated nucleocapsid phosphoprotein of rinderpest virus, the Bowman-Birk type of proteinase inhibitors, and the metallothionein superfamily of cysteine rich chelating proteins have been identified. Comparison of the TGEV polymerase sequence with that of other RNA viruses revealed high sequence homology with the A-E domains of the palm subdomain of nucleic acid polymerases.

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Section: Introductionmentioning

confidence: 99%

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et al. 2001

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“…Thus positive-strand DI RNA synthesis, from the expressed negative-strand DI RNA transcripts, is blocked at the ds RNA structure present downstream of the leader sequence in coexpressing cells. Nascent leader sequence containing subgenomic mRNAs elongate on genome-length replicative intermediate RNA, containing the genome-length negativestrand template Mizutani et al, 2000). As most of coronavirus negative-strand RNAs appear to be present in a ds RNA form (Lin et al, 1994;Sawicki and Sawicki, 1986), genome-length negative-strand RNA, which is a template for MHV subgenomic mRNA synthesis Baric et al, 1983;Mizutani et al, 2000), also most probably exists in a ds RNA form.…”

Section: Mechanism Of Leader Switching In Cells Expressing Negative-smentioning

confidence: 99%

“…Nascent leader sequence containing subgenomic mRNAs elongate on genome-length replicative intermediate RNA, containing the genome-length negativestrand template Mizutani et al, 2000). As most of coronavirus negative-strand RNAs appear to be present in a ds RNA form (Lin et al, 1994;Sawicki and Sawicki, 1986), genome-length negative-strand RNA, which is a template for MHV subgenomic mRNA synthesis Baric et al, 1983;Mizutani et al, 2000), also most probably exists in a ds RNA form. Although further studies are required to clarify the mechanism of the joining of leader sequence to the 5Ј end of subgenomic mRNA during subgenomic mRNA synthesis, it was proposed that the leader sequence is used as a primer for subgenomic mRNA synthesis (leader-primed transcription) Lai et al, 1984b).…”

Section: Mechanism Of Leader Switching In Cells Expressing Negative-smentioning

confidence: 99%

“…Genomic-size and subgenomic-size negative-strand RNAs are present in coronavirus-infected cells in amounts that are significantly lower than the amounts of corresponding positive-strand RNAs (Sethna et al, 1989). Two independent studies showed that nascent leader sequence-containing MHV subgenomic mRNAs are elongating on a genomic-length replicative-intermediate RNA containing a genomiclength negative-strand RNA late in infection Mizutani et al, 2000). The genomic-length negative-strand RNA also serves as a template for subgenomic mRNA synthesis early in infection .…”

Section: Introductionmentioning

confidence: 99%

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Enhanced Accumulation of Coronavirus Defective Interfering RNA from Expressed Negative-Strand Transcripts by Coexpressed Positive-Strand RNA Transcripts

2001

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Expression of negative-strand murine coronavirus mouse hepatitis virus (MHV) defective interfering (DI) RNA transcripts in MHV-infected cells results in the accumulation of positive-strand DI RNAs (M. Joo et al., 1996, J. Virol. 70, 5769-5776). However, the expressed negative-strand DI RNA transcripts are poor templates for positive-strand DI RNA synthesis. The present study demonstrated that DI RNA accumulation from the expressed negative-strand DI RNA transcripts in MHV-infected cells was enhanced by the coexpression of complementary RNA transcripts that correspond to the 5' region of positive-strand DI RNA. The positive-strand RNA transcripts corresponding to the 5' end-most 0.7-2.0 kb DI RNA had a similar enhancement effect. The coexpressed positive-strand RNA transcripts lacking the leader sequence or those containing only the leader sequence failed to demonstrate this enhancement effect, demonstrating that the presence of the leader sequence in the coexpressed positive-strand RNA transcripts was necessary, but not sufficient, for the enhancement of DI RNA accumulation from the coexpressed negative-strand DI RNA transcripts. Negative-strand DI RNA transcripts that were coexpressed with the partial-length positive-strand RNA transcripts were no more stable than those expressed alone, suggesting that a higher stability of the expressed negative-strand RNA transcripts was an unlikely reason for the higher DI RNA accumulation in cells coexpressing two complementary DI RNA transcripts. Sequence analyses unexpectedly demonstrated that the leader sequence of the majority of accumulated DI RNAs switched to helper virus derived leader sequence, suggesting that enhancement of DI RNA accumulation was mediated by the efficient utilization of helper virus derived leader sequence for DI RNA synthesis. Furthermore, our data suggested that this leader switching, a type of homologous RNA-RNA recombination, occurred during positive-strand DI RNA synthesis and that MHV positive-strand RNA synthesis mechanism may have a preference toward recognizing double-stranded RNA structures over single-stranded negative-strand RNA to produce positive-strand DI RNAs.

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“…The SARSCoV genome is about 30 kb long. Differential transcription of its genes generates a gradient of subgenomic RNAs (sgRNA) with a common 3Ј end (2,(4)(5)(6). This implies that genes at the 3Ј end should be transcribed at high levels and that sgRNA species produced by transcription of these genes might be the most abundant during cell infection.…”

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confidence: 99%