Fig. 4: A critical role for dimerization of Unkempt.
From: Regulation by the RNA-binding protein Unkempt at its effector interface
A A representative AlphaFold prediction of the coiled-coil homodimer formed by two copies of the C-terminal segment of UNK (UNKC; residues 637–810). All 25 predictions were identical, bar some minor variations in the C-terminal loop regions. The C-alpha atoms of the key hydrophobic residues stabilizing the coiled coil are shown as yellow spheres. Predictions of the homodimer full-length UNK protein are shown in Supplementary Fig. 5B. B–D Mass photometry analyses of Strep-tagged UNKC or UNKFULL (B), UNKC E6 or UNKC E8 (C), CNOTMINI or in complex with UNKFULL (D) (n = 2). Mutations in UNKC E6 and UNKC E8 are listed in (C). The calculated molecular weights are 62.5 kDa for UNKC monomer, 125 kDa for UNKC dimer, 90.5 kDa for UNKFULL monomer, and 181 kDa for UNKFULL dimer. The observed mean molecular weights are 126 kDa for UNKC, 62 kDa and 125 kDa for UNKC E6, 58 kDa for UNKC E8, 189 kDa for UNKFULL, 326 kDa for CNOTMINI, and 186 kDa, 321 kDa, and 512 kDa for the mixture of UNKFULL and CNOTMINI. E Morphologies of cells inducibly co-expressing UNKWT or UNKE8 and GFP compared to GFP-only expressing cells at 48 h of incubation with Dox (n = between 44 and 59 GFP-expressing cells per cell line). Data are presented as mean ± SD. Statistical significance was determined using Student’s two-tailed t-test with *p = 1.0 × 10−23; ns not significant.
