Human ZC3H14 isoforms 1–3 are localized to the nucleus of N2A cells. (A) Domain alignment of S. cerevisiae Nab2, D. melanogaster dNab2, and human ZC3H14 isoforms 1–3 and isoform 4. The conserved N-terminal PWI-like fold, Q-rich, RGG/predicted classical nuclear localization signal (cNLS), and C-terminal tandem CCCH zinc finger RNA-binding motif (ZnF CCCH) domains are indicated. The zinc finger domains of human ZC3H14 and fly dNab2 share significant homology with 48% amino acid similarity in the zinc fingers as determined by analysis with Clustal W (Larkin et al. 2007). The location of the Arginine-to-termination codon (R154Ter) mutation that eliminates isoforms 1–3 in intellectual disability patients is shown as is the location for the Alternate exons that are included/excluded to generate ZC3H14 isoforms 1–3. The PWI-like domain used to raise polyclonal ZC3H14 antibodies, which recognize ZC3H14-iso1-3, (Leung et al. 2009) is also indicated (Antibody binding region). (B) ZC3H14 localization in cultured N2A cells was assessed using an antibody directed against the PWI-like domain of ZC3H14 (α-ZC3H14) and thus isoforms 1–3 are visualized (green). Cells are costained with the RNA speckle marker SC-35 (α-SC-35) to indicate the position of nuclear speckles (red). Corresponding Hoescht staining (blue) is shown to indicate the position of the nucleus. A phase image is also shown.
