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Figure 1.
(a) Schematic representation of the domain organization of
DBP5 with the domain boundaries of the protein constructs used
in this study. The RecA-like domains are colored blue and the
N-terminal region is shown in orange. (b) Protein precipitations
with biotinylated single-stranded RNA identify the RNA binding
region of human DBP5 for structural studies. Purified proteins
were mixed with 5' end–biotinylated 20-mer single-stranded RNA
and incubated with or without AMPPNP or ADP, as indicated.
Proteins mixtures before (input, 17% of the total) and after
coprecipitation (precipitate) were separated on a 15% (w/v)
acrylamide SDS-PAGE and visualized using Coomassie stain. The
molecular weight standards are shown in lane 1. DBP5 constructs
are described in a. NUP214   C
corresponds to residues 1–405 (human sequence). (c) Structure
of DBP5 N
(light blue) bound to AMPPNP (yellow) and single-stranded RNA
(black). The magnesium ion at the ATP binding site is shown in
magenta. Six ordered nucleotides of a single-stranded poly-U RNA
are present in the 2.2-Å resolution structure. The N- and
C-terminal residues of DBP5 visible in the electron density are
indicated. The N-terminal region of DBP5 (residues 75–92,
orange) folds into a short  -helix.
This and all other ribbon diagrams in the manuscript were
generated using PyMol (http://www.pymol.org).
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