Figure 6.

(A) Identification of MCPIP1-triggered cleavage sites in the mIL-6_82–106 stem-loop RNA structure. Nt sequences and structures created during MCPIP1-induced cleavage are illustrated. Mapping of the cleavage sites based on the RNase assay results, intermediates and the most significant subsequent degradation products are presented. (B) Visualization of the stoichiometry of the MCPIP1 interaction with stem-loops. Schematic cartoon representation of the ternary complex model. The size exclusion chromatography results showed that PIN and PIN-ZF were monomeric and suggest that full-length MCPIP1 most frequently occurs as a dimer in native condition. Stoichiometry of the MCPIP1 - RNA interaction was based on the size exclusion chromatography results and the results from affinity determination assays where the sequential binding model were used. Thus, for full-length MCPIP1, we proposed a sequential binding model: oligo + MCPIP1_Dimer + MCPIP1_dimer $\rightleftarrows$ oligo-MCPIP1_dimer + MCPIP1_dimer $\rightleftarrows$ oligo-MCPIP1_tetramer. The presented dissociations constants of the complexes were estimated based on the affinity determination assays shown in Table 2.