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Formation of a pseudoknot in the conserved RNA core domain in theribonucleoprotein human telomerase is required for function. In vitroexperiments show that the pseudoknot (PK) is in equilibrium with an extendedhairpin (HP) structure. We use molecular simulations of a coarse-grained model,which reproduces most of the salient features of the experimental meltingprofiles of PK and HP, to show that crowding enhances the stability of PKrelative to HP in the wild type and in a mutant associated with dyskeratosiscongenita. In monodisperse suspensions, small crowding particles increase thestability of compact structures to a greater extent than larger crowders. Ifthe sizes of crowders in a binary mixture are smaller than the unfolded RNA,the increase in melting temperature due to the two components is additive. In aternary mixture of crowders that are larger than the unfolded RNA, which mimicsthe composition of ribosome, large enzyme complexes and proteins in E. coli,the marginal increase in stability is entirely determined by the smallestcomponent. We predict that crowding can restore partially telomerase activityin mutants, which dramatically decrease the PK stability.

Crowding Promotes the Switch from Hairpin to Pseudoknot Conformation in Human Telomerase RNA