One-dimensional-NMR data suggest that the peptide changes the population distribution of possible RNA structures to favor an annealing-prone RNA conformation, thereby increasing the fraction of colliding RNA molecules that successfully anneal.Īs with proteins, the folding of RNA molecules into their native and functional structure is a non-trivial problem ( 1, 2). Additionally, we found that Tat(44–61) drives the RNA annealing reaction via entropic rather than enthalpic terms. Mutagenesis of the peptide illustrated the dominant role of positively charged amino acids in RNA annealing-both the overall charge of the molecule and a precise distribution of basic amino acids within the peptide are important. The activity of the peptide is strongly regulated by mono- and divalent cations which hints at the importance of electrostatic interactions between RNA and peptide. In order to study the mechanism of protein-facilitated acceleration of annealing we selected a short peptide, HIV-1 Tat(44–61), which accelerates the reaction efficiently. A variety of proteins have been shown to accelerate RNA/RNA annealing but their mode of action is still mainly uncertain. The annealing of nucleic acids to (partly) complementary RNA or DNA strands is involved in important cellular processes.
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