Efficient stimulation of site-specific ribosome frameshifting by antisense oligonucleotides

Abstract

Evidence is presented that morpholino, 2-O-methyl, phosphorothioate, and RNA antisense oligonucleotides can direct sitespecific −1 translational frameshifting when annealed to mRNA downstream from sequences where the P-and A-site tRNAs are both capable of re-pairing with −1 frame codons. The efficiency of ribosomes shifting into the new frame can be as high as 40%, determined by the sequence of the frameshift site, as well as the location, sequence composition, and modification of the antisense oligonucleotide. These results demonstrate that a perfect duplex formed by complementary oligonucleotides is sufficient to induce high level −1 frameshifting. The implications for the mechanism of action of natural programmed translational frameshift stimulators are discussed.

The global structures of a wild-type and poorly functional plant luteoviral mRNA pseudoknot are essentially identical

“…These disparate findings, taken together, indicate that the impact of specific interactions at the helical junction are strongly context dependent, and structural/functional rules that emerge from investigation of a specific frameshift signal may not be generally applicable to systems. Indeed, there may be multiple mechanistic paths the ribosome can take to achieve high levels of À1 frameshifting since antisense oligonucleotides capable of base pairing to a site just downstream of the slip site have recently been shown to induce significant levels of frameshifting in vitro (Howard et al 2004).…”

Augmented genetic decoding: global, local and temporal alterations of decoding processes and codon meaning

“…mRNA interacts with various cellular components, and these interactions may alter the stimulatory properties of particular structures or sequences. Both protein molecules 140 and nucleic acid molecules 117,141,142 have been shown to modulate frameshifting in trans. The dependence of frameshifting on stimulatory elements, as well as the responsiveness to cellular conditions, provides translation with a powerful regulatory mechanism.…”

Programmed ribosomal frameshifting in HIV-1 and the SARS–CoV

“…Thus, frameshifting at stem-loop structures, like at pseudoknots, is likely to be governed by the rate at which the structure is unfolded. It has recently been discovered that simply annealing an oligonucleotide downstream of a slippery sequence can under certain circumstances promote efficient frameshifting, at least in in vitro translation systems (Howard et al, 2004;Olsthoorn et al, 2004). In this situation, the ribosome encounters a double-stranded region conformationally distinct from a stem-loop, yet similarly, the rate of unwinding of the structure is likely to be the key determinant of frameshifting efficiency.…”