+ 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

An in vitro system for the leader primed transcription of coronavirus messenger rnas



An in vitro system for the leader primed transcription of coronavirus messenger rnas



EMBO (European Molecular Biology Organization) Journal 9(12): 4173-4180



We have developed an in vitro transcription system which can utilize exogenous leader RNA for mouse hepatitis virus (MHV) 'leader-primed' mRNA transcription. Cytoplasmic extracts containing viral proteins and template RNA were prepared by lysolecithin permeabilization of MHV-infected cells. Synthetic leader RNA which differed in sequence from the endogenous leader RNA was added to the extracts and demonstrated to be incorporated into MHV mRNAs. Irrespective of the size of leader RNAs added, the exogenous leader RNA was joined to the endogenous mRNA at the same site, which corresponds to a UCUAA pentanucleotide repeat region. Only leader RNAs containing the pentanucleotide sequences could be utilized for transcription. Mismatches between the intergenic site and the exogenous leader sequence within the pentanucleotide repeat region were corrected in the in vitro system. This in vitro system thus established a novel mechanism of leader-primed transcription using exogenous RNA in trans, and suggests the involvement of a specific ribonuclease activity during coronavirus mRNA synthesis.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 007011568

Download citation: RISBibTeXText


Related references

Characterization of leader-related small RNAs in coronavirus-infected cells: further evidence for leader-primed mechanism of transcription. Virus Research 3(1): 19-33, 1985

An in vitro system for the leader-primed transcription of coronavirus mRNAs. Embo Journal 9(12): 4173-4179, 1990

An in vitro system for the leader-primed transcription of coronavirus mRnas. The Embo Journal 9(12): 4173-4179, 1990

Characterization of leader related small rna species in coronavirus infected cells further evidence for leader primed mechanism of transcription. Virus Research 3(1): 19-34, 1985

Coronavirus leader-RNA-primed transcription: an alternative mechanism to RNA splicing. Bioessays 5(6): 257-260, 1986

Mechanism of coronavirus rna transcription anti sense leader rna inhibits messenger rna transcription. Journal of Cellular Biochemistry Suppl. (10 Part D): 295, 1986

Murine coronavirus 5'-end genomic RNA sequence reveals mechanism of leader-primed transcription. Advances in Experimental Medicine and Biology 218: 73-81, 1987

The 5'-end sequence of the murine coronavirus genome: implications for multiple fusion sites in leader-primed transcription. Virology 156(2): 321-330, 1987

Leader sequences of murine coronavirus messenger rna can be freely reassorted evidence for the transcriptional role of free leader rna. Journal of Cellular Biochemistry Suppl. (10 Part D): 294, 1986

Multiple leader RNAs and messenger RNAs are transcribed from the La Crosse virus small genome segment. Cell 33(3): 791-799, 1983

Short leader sequences may be transferred from small rnas to pre mature messenger rnas by trans splicing in euglena. EMBO (European Molecular Biology Organization) Journal 10(9): 2621-2626, 1991

Leader sequences of murine coronavirus mRNAs can be freely reassorted: evidence for the role of free leader RNA in transcription. Proceedings of the National Academy of Sciences of the United States of America 83(12): 4204-4208, 1986

Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs. Nucleic Acids Research 12(18): 7057-7070, 1984

In vitro transcription and translational efficiency of chimeric SP6 messenger RNAs devoid of 5' vector nucleotides. Nucleic Acids Research 16(10): 4483-4498, 1988

Subgenomic RNAs with nucleotide sequences derived from RNAs 1 and 2 of cucumber mosaic virus can act as messenger RNAs in vitro. Virology 142(1): 144-157, 1985