Folding pathways of large RNAs are poorly understood.
We have addressed this question by hybridizing in vitro
transcripts, which varied in size, to an array of antisense
oligonucleotides. All transcripts included a common sequence
and all but one shared the same start-point; the other
had a small deletion of the 5′ end. Minimal free
energy calculations predicted quite different folds for
these transcripts. However, hybridization to the array
showed predominant features that were shared by transcripts
of all lengths, though some oligonucleotides that hybridized
strongly to the short transcripts gave weak interaction
with longer transcripts. A full-length RNA fragment that
had been denatured by heating and allowed to cool slowly
gave the same hybridization result as a shorter transcript.
Taken together, these results support theories that RNA
folding creates local stable states that are trapped early
in the transcription or folding process. As the transcript
elongates, interactions are added between regions that
are transcribed early and those transcribed late. The method
here described helps in identifying regions in the transcripts
that take part in long-range interactions.