1.2. CENTRO DE RECREACIÓN CULTURAL
1.2.7. Elementos del diseño organizacional para el centro de recreación
relative to the structures these sequences form in the monomeric initial state (illustrated in Table 2.1).
Structures for PAL1 and PAL2 in the monomer state were obtained using strong and moderate SHAPE reactivities to constrain single stranded nucleotides15. PAL1 stability was calculated for nts 205–227. PAL2 stability was estimated for nts 244-299, as the mean folding free energy for three postulated15 interconverting structures in the 231-315 domain
and subtracting the free energy of the anchoring duplex (nts 231-243/300-315). Stabilities for the PAL1 and PAL2 intermolecular duplexes in the dimer state were estimated using the
bimolecular folding mode in RNAstructure34.
2.5.6 Incremental dissociation constants for individual dimerization steps. For the dimerization step involving SL1-SL2 domain interactions, the dimerization constant K1
was measured directly as the affinity of the Stb-Del mutant. For steps involving SL1-SL2 interactions plus either PAL1 or PAL2 formation, K2 and K2' were calculated from Kdimer for
the PAL2Del and PAL1Stb mutants, respectively, as Kdimer = K1K2. For the third step, K3 and
K3' were calculated from Kdimer for dimerization of the native MiDAS, Kdimer = K1K2K3. Free
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