Pseudoknots in RNA secondary structures: representation, enumeration, and prevalence.

A number of non-coding RNA are known to contain functionally important or conserved pseudoknots. However, pseudoknotted structures are more complex than orthodox, and most methods for analyzing secondary structures do not handle them. I present here a way to decompose and represent general secondary structures which extends the tree representation of the stem-loop structure, and use this to analyze the frequency of pseudoknots in known and in random secondary structures. This comparison shows that, though a number of pseudoknots exist, they are still relatively rare and mostly of the simpler kinds. In contrast, random secondary structures tend to be heavily knotted, and the number of available structures increases dramatically when allowing pseudoknots. Therefore, methods for structure prediction and non-coding RNA identification that allow pseudoknots are likely to be much less powerful than those that do not, unless they penalize pseudoknots appropriately.