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. 2013 Sep;19(9):1238–1252. doi: 10.1261/rna.039255.113

FIGURE 6.

FIGURE 6.

RNA-binding influences Dicer localization. (A) Transfection of HeLa cells with the C-terminal fragment of Dicer (RNase III domains plus dsRBD). The wild-type (wt) construct fails to localize to the nucleus. When E1564K is introduced into RNase IIIa catalytic core (RNase IIIa mut), nuclear accumulation can be observed. A similar mutation (E1813K) in the catalytic core of RNase IIIb (RNase IIIb mut) also leads to partial localization in the nucleus, although the effect is much weaker compared to RNase IIIa mut. The double mutation E1564K/E1813K showed strong nuclear accumulation. (B) Transfection of HeLa cells with full-length Dicer constructs. Wild-type (wt) Dicer localizes to the cytoplasm. Introduction of mutations E1564K or E1813K into the RNase III domains either individually (RNase IIIa mut and RNase IIIb mut) or together (RNase IIIa & b mut) showed cytoplasmic staining similar to wt protein. Representative images are shown from a minimum of three independent experiments. Bar, 20 µm. (C) Schematic representation of human Dicer with individual protein domains indicated by different colors and the position of each RNase III mutation shown.