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In vitro-transcribed biotinylated human SRP RNA was incubated with HNE in the presence or absence of recombinant RanQ69LGTP, a constitutively active mutant form of Ran loaded with GTP, the RNA was then pulled down with streptavidin beads, and the precipitated protein was analyzed by Western blotting (Fig. 5).
The active GTP-bound form of Ran (Ran-GTP) is produced on the chromatin and a Ran-GTP gradient centred on chromosomes is generated (for review see [7]).
(D ) Slx9 specifically interacts with the GTP-bound form of Ran.
Pre-miRNAs interact with exportin-5 and the GTP-bound form of Ran in the nucleus.
Surprisingly, although Slx9 did not interact with Crm1 in vitro, it interacts specifically with the GTP-bound form of Ran.
RCC1 is a guanine exchange factor for the Ran GTPase (or RanGEF) that establishes a gradient of the GTP bound form of Ran around chromatin.
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