Nucleic Acids Research, 2006, Vol. 34, No. 16 4605 (a) (b) (c) Figure 6. Analysis of regions responsible for subcellular localization of DDX47. (a) Serial deletion constructs of EGFP-fused DDX47 were used to transfect BHK21 cells. Subcellular localization of all constructs was determined by fluorescence microscopy. Representative fluorescence images are shown. Interaction of DDX47 deletion constructs with NOP132 was determined using two different yeast two-hybrid read-outs, a chromogenic b-galactosidase assay and a colony growth assay, and the results are shown at the far right. Domains: DEADc, CDD accession no. cd00268 (38); HELICc, CDD accession no. cd00079. Abbreviations: N, nucleus/nucleolus; C, cytoplasm; NES, nuclear export signal; NLS, nuclear localization signal; +, yeast two-hybrid interaction between the DDX47 deletion construct and NOP132; , no interaction. Hatched box indicates the NOP132-interacting region. N, nucleus; C, cytoplasm. (b) DDX47 amino acid sequence is shown indicating possible NES and NLS regions (underlined) and motifs (bold). (c) Sequence alignment of the DDX47 and HELICc motifs. Conserved RNA helicase DEAD-box motifs IV, V, and VI are indicated (48). Two dots indicate identity and a single dot indicates chemically similar amino acids.