Effect of intergenic consensus sequence flanking sequences on coronavirus transcription

Shinji Makino, Myungsoo Joo

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Insertion of a region, including the 18-nucleotide-long intergenic sequence between genes 6 and 7 of mouse hepatitis virus (MHV) genomic RNA, into an MHV defective interfering (DI) RNA leads to transcription of subgenomic DI RNA in helper virus-infected cells (S. Makino, M. Joo, and J. K. Makino, J. Virol. 66:6031-6041, 1991). In this study, the subgenomic DI RNA system was used to determine how sequences flanking the intergenic region affect MHV RNA transcription and to identify the minimum intergenic sequence required for MHV transcription. DI cDNAs containing the intergenic region between genes 6 and 7, but with different lengths of upstream or downstream flanking sequences, were constructed. All DI cDNAs had an 18-nucleotide-long intergenic region that was identical to the 3′ region of the genomic leader sequence, which contains two UCUAA repeat sequences. These constructs included 0 to 1,440 nucleotides of upstream flanking sequence and 0 to 1,671 nucleotides of downstream flanking sequence. An analysis of intracellular genomic DI RNA and subgenomic DI RNA species revealed that there were no significant differences in the ratios of subgenomic to genomic DI RNA for any of the DI RNA constructs. DI cDNAs which lacked the intergenic region flanking sequences and contained a series of deletions within the 18-nucleotide-long intergenic sequence were constructed to determine the minimum sequence necessary for subgenomic DI RNA transcription. Small amounts of subgenomic DI RNA were synthesized from genomic DI RNAs with the intergenic consensus sequences UCUAAAC and GCUAAAC, whereas no subgenomic DI RNA transcription was observed from DI RNAs containing UCUAAAG and GCTAAAG sequences. These analyses demonstrated that the sequences flanking the intergenic sequence between genes 6 and 7 did not play a role in subgenomic DI RNA transcription regulation and that the UCUAAAC consensus sequence was sufficient for subgenomic DI RNA transcription.

Original languageEnglish (US)
Pages (from-to)3304-3311
Number of pages8
JournalJournal of Virology
Volume67
Issue number6
StatePublished - Jun 1993
Externally publishedYes

Fingerprint

Coronavirus
Intergenic DNA
Coronavirinae
consensus sequence
Consensus Sequence
transcription (genetics)
RNA
intergenic DNA
Murine hepatitis virus
nucleotides
Nucleotides
genomics
Complementary DNA
Helper Viruses
Genes
genes

ASJC Scopus subject areas

  • Immunology

Cite this

Effect of intergenic consensus sequence flanking sequences on coronavirus transcription. / Makino, Shinji; Joo, Myungsoo.

In: Journal of Virology, Vol. 67, No. 6, 06.1993, p. 3304-3311.

Research output: Contribution to journalArticle

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