Effect of single-base substitutions in the central domain of virus-associated RNA I on its function.

AUTOR(ES)

Rahman, A

RESUMO

Adenoviruses use virus-associated RNA I (VAI RNA) to counteract the cellular antiviral response mediated by the interferon-induced, double-stranded-RNA-activated protein kinase PKR. VAI RNA is a highly structured small RNA which consists of two long duplex regions connected at the center by a complex, short stem-loop. This short stem-loop and the adjacent base-paired regions, referred to as the central domain, bind to PKR and inactivate it. Currently it is not known whether binding of VAI RNA to PKR is dependent solely on the secondary (and tertiary) structure of the central domain or whether nucleotide sequences in the central domain are also critical for this interaction. To address this question, 54 VAI mutants with single-base substitution mutations in the central domain of the RNA were constructed, and their capacities to inhibit the autophosphoryation of PKR in vitro were determined. It was found that although about half of the mutants inhibited PKR activity as efficiently as the wild type, a significant number of mutants lost the inhibitory activity substantially, without a perceptible change in their secondary structures. These results indicate that, in addition to secondary structure, at least some nucleotides in the central domain may be critical for the efficient function of VAI RNA.

Documentos Relacionados

• Effects of single-base substitutions within the Acanthamoeba castellanii rRNA promoter on transcription and on binding of transcription initiation factor and RNA polymerase I.
• Enzymatic techniques for the isolation of random single-base substitutions in vitro at high frequency.
• Enzymatic techniques for the isolation of random single-base substitutions in vitro at high frequency
• Mutational analysis of the central domain of adenovirus virus-associated RNA mandates a revision of the proposed secondary structure.
• Non-cross-linking gold nanoparticle aggregation as a detection method for single-base substitutions