Comparative study of the effects of heptameric slippery site composition on -1 frameshifting among different eukaryotic systems.
Studies of programmed -1 ribosomal frameshifting (-1 PRF) have been approached over the past two decades by many different laboratories using a diverse array of virus-derived frameshift signals in translational assay systems derived from a variety of sources. Though it is generally acknowledged that both absolute and relative -1 PRF efficiency can vary in an assay system-dependent manner, no methodical study of this phenomenon has been undertaken. To address this issue, a series of slippery site mutants of the SARS-associated coronavirus frameshift signal were systematically assayed in four different eukaryotic translational systems. HIV-1 promoted frameshifting was also compared between Escherichia coli and a human T-cell line expression systems. The results of these analyses highlight different aspects of each system, suggesting in general that (1) differences can be due to the assay systems themselves; (2) phylogenetic differences in ribosome structure can affect frameshifting efficiency; and (3) care must be taken to employ the closest phylogenetic match between a specific -1 PRF signal and the choice of translational assay system.
Department of Cell Biology and Molecular Genetics, Microbiology Building, Room 2135, University of Maryland, College Park, Maryland 20742, USA.
Ewan P
EP
Plant
Jonathan D
JD
Dinman
D001483:Base Sequence; D002460:Cell Line; D003062:Codon; D004926:Escherichia coli / Q000235:genetics; D018965:Frameshifting, Ribosomal; D015497:HIV-1; D006801:Humans; D016297:Mutagenesis, Site-Directed; D009841:Oligonucleotides; D010957:Plasmids; D013601:T-Lymphocytes / Q000378:metabolism
D003062:Codon; D009841:Oligonucleotides