Structure of a hexanucleotide RNA hairpin loop conserved in ribosomal RNAs

The structure of 5'-pppGGAC(GUAAUA)GUCC has been deduced from NMR data. The six-nucleotide hairpin loop is highly conserved in large subunit ribosomal RNA and confers unusual stability on RNA hairpins. Standard assignment methods, including through-bond strategies for backbone protons, were used to assign all P-31 resonances and all but two of the non-exchangeable protons. The model calculated by restrained molecular dynamics shows that the loop adopts a specific structure stabilized by five non-canonical hydrogen bonds. An edge-to-edge G . A pair (hydrogen bonds G5 N3-A10 NH6, G5 H2-A10 N7 and A10 NH6-G5 O2') closes the hairpin loop. A tuans 5' P-O bond at A7 reverses the direction of the backbone and is stabilized by two base-backbone hydrogen bonds (U6 NH3-U9 OP and U6 O2'-A8 N7). The only unstructured part of the loop is U9, which is completely unstacked and also the only phylogenetically variable position. The U6-A7-A8 ''U-turn'' reproduces hydrogen bonds and backbone torsion seen in the tRNA anticodon loop and T Psi C loop, and in an internal loop of the hammerhead ribozyme. G . A is a common closing mismatch in ribosomal RNA hairpin loops. The U-turn and G . A pair found in this hexaloop structure may therefore be common structural units of RNA hairpin loops. (C) 1996 Academic Press Limited