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. 2023 Mar 15;133(6):e165245. doi: 10.1172/JCI165245

Figure 3. 5′-L deletions alter dimerization propensities and NC-binding properties.

Figure 3

(A) Secondary structure of the HIV-1 NL4-3 gRNA 5′-L with patient deletions indicated in purple (d22) and orange (d21), with a region of overlap within the splice donor (SD) shown in red. High-affinity binding sites with endothermic contribution on ITC binding within the psi hairpin are indicated with blue text. Gray text indicates the portion of AUG truncated to better study the dimer and its initial binding sites by ITC. (B) Concentration-dependent dimerization assays of the full leader show that the WT and d21 constructs maintain similar dimerization propensities, while the d22 variants exhibit reduced dimerization. (C–E) ITC isotherms for the truncated dimeric 5′-L titrated with low protein-to-RNA ratios. WT exhibits previously described initial binding with an endothermic contribution (C) that is not seen for the d22 (D) or d21 (E) 5′-L constructs.