The nucleotide sequence of satellite tobacco necrosis virus strain C and helper-assisted replication of wild-type and mutant clones of the virus

DH Bringloe, AP Gultyaev, M Pelpel, CW Pleij and RH Coutts Biology Department, Imperial College of Science, Technology and Medicine, London, UK. The complete nucleotide sequence of satellite tobacco necrosis virus strain C (STNV-C) was determined. The genome has a similar overall organization to two...

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Bibliographic Details
Published in:Journal of general virology 1998-06, Vol.79 (6), p.1539-1546
Main Authors: Bringloe, DH, Gultyaev, AP, Pelpel, M, Pleij, CW, Coutts, RH
Format: Article
Language:English
Subjects:
Base Sequence
DNA, Viral
Helper Viruses - genetics
Helper Viruses - physiology
Molecular Sequence Data
Mutation
Nicotiana - virology
Nucleic Acid Conformation
Plants, Toxic
RNA, Viral - chemistry
Satellite Viruses - genetics
Satellite Viruses - physiology
Sequence Analysis, RNA
Virus Replication
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Summary:DH Bringloe, AP Gultyaev, M Pelpel, CW Pleij and RH Coutts Biology Department, Imperial College of Science, Technology and Medicine, London, UK. The complete nucleotide sequence of satellite tobacco necrosis virus strain C (STNV-C) was determined. The genome has a similar overall organization to two STNV isolates studied previously but differs significantly from them in the secondary structure of the translated and untranslated regions (UTRs). STNV-C RNA is naturally uncapped and contains 1221 nt: 101 nt in the 5' UTR, 606 nt in the capsid protein (CP) coding region and 514 nt in the 3' UTR. Using the known sequences of STNV-C and tobacco necrosis virus strain D (TNV-D) RNAs, full-length cDNA clones of both RNAs were constructed. Synthetic transcripts derived from STNV-C cDNA clones only replicated in plants and protoplasts when co-inoculated with TNV-D transcripts. A number of mutant clones in both the 3' and the 5' STNV-C RNA UTRs were constructed which disrupted putative cis-acting elements recognized by helper virus polymerase. Deletion analysis revealed an essential requirement of all 3' and 5' proximal sequences in the STNV-C UTRs for replication. However, an internal region in the 3' UTR could be deleted without loss of infectivity. Likewise, the entire STNV-C CP-encoding region could be deleted and replaced with a marker gene of a similar size without loss of transcript accumulation in plants.
ISSN:0022-1317
1465-2099
DOI:10.1099/0022-1317-79-6-1539