Pseudoknot in the central domain of small subunit ribosomal RNA is essential for translation

Phylogenetic comparison of rRNA sequences has suggested that a pseudoknot structure exists in the central domain of small-subunit rRNA. In Escherichia coli 16S rRNA, this pseudoknot would form when positions 570 and 571 pair with positions 865 and 866. Mutations were introduced into this pseudoknot at the phylogenetically invariant nucleotides U571 and A865. Single mutations of U to A at 571 or A to U at 865 dramatically altered the structural stability of the 30S subunit and also impaired the function of the subunit in translation. When the mutations were combined to create a compensatory pairing, the normal structure of the 30S subunit was restored, and the function of the mutant subunit in translation returned to wild-type levels. These results demonstrate the existence of a higher order structure in rRNA that directly affects the folding of the 30S subunit. Given the position of this structure in the three-dimensional model of the small subunit and the additional interactions that are likely to form in the same rRNA region, the central domain pseudoknot appears to contribute to a complex structure of rRNA that controls the conformational state of the ribosome.