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Methylation of the exocyclic amino group of guanine is a relatively common modification in rRNA and tRNA. Single methylation ( N 2  -methylguanosine, m 2 G) is the second most frequently encountered nucleoside analog in  Escherichia coli  rRNAs. The most prominent case of dual methylation ( N 2 ,N 2  -dimethylguanosine, m 2  2 G) is found in the majority of eukaryotic tRNAs at base pair m 2  2 G26:A44. The latter modification eliminates the ability of the  N 2   function to donate in hydrogen bonds and alters its pairing behavior, notably vis-à-vis C. Perhaps a less obvious consequence of the  N 2 ,N 2  -dimethyl modification is its role in controlling the pairing modes between G and A. We have determined the crystal structure of a 13-mer RNA duplex with central tandem m 2  2 G:A pairs. In the structure both pairs adopt an imino-hydrogen bonded, pseudo-Watson–Crick conformation. Thus, the sheared conformation frequently seen in tandem G:A pairs is avoided due to a potential steric clash between an  N 2  -methyl group and the major groove edge of A. Additionally, for a series of G:A containing self-complementary RNAs we investigated how methylation affects competitive hairpin versus duplex formation based on UV melting profile analysis.

Effects of N2,N2-dimethylguanosine on RNA structure and stability: Crystal structure of an RNA duplex with tandem m22G:A pairs