An astrovirus frameshift signal induces ribosomal frameshifting in vitro

Expression of the astrovirus RNA-dependent RNA polymerase has been hypothesized to be regulated by (-1) ribosomal frameshifting. Sequence analysis of the 70 nucleotide region between open reading frames 1a and 1b indicates the presence of a shifty heptamer consensus sequence and downstream sequences...

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Bibliographic Details
Published in:Archives of virology 1995-01, Vol.140 (6), p.1127-1135
Main Authors: LEWIS, T. L, MATSUI, S. M
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Biological and medical sciences
Brief Report
Capsid - genetics
Capsid Proteins
Cloning, Molecular
DNA, Viral
Frameshift Mutation
Fundamental and applied biological sciences. Psychology
Genetics
Humans
Macaca mulatta
Mamastrovirus - genetics
Microbiology
Molecular Sequence Data
Ribosomes
Rotavirus - genetics
Virology
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Summary:Expression of the astrovirus RNA-dependent RNA polymerase has been hypothesized to be regulated by (-1) ribosomal frameshifting. Sequence analysis of the 70 nucleotide region between open reading frames 1a and 1b indicates the presence of a shifty heptamer consensus sequence and downstream sequences that may be needed for ribosomal frameshifting. We constructed four astrovirus cassettes that spanned this region and inserted each into the rhesus rotavirus VP4 gene. The constructs were expressed in an in vitro system, and products were immunoprecipitated by rotavirus amino and carboxy terminal-specific monoclonal antibodies. Ribosomal frameshifting, at an efficiency of 6-7%, was demonstrated in all constructs containing the shifty heptamer and stem-loop. Deletion of the downstream sequence potentially involved in pseudoknot formation did not affect frameshifting efficiency. However, deletion of the shifty heptamer resulted in no detectable frameshift activity.
ISSN:0304-8608
1432-8798
DOI:10.1007/BF01315421