Author: Aaron S. Mendez; Carolin Vogt; Jens Bohne; Britt A. Glaunsinger
Title: Site specific target binding controls RNA cleavage efficiency by the Kaposi’s sarcoma-associated herpesvirus endonuclease SOX Document date: 2018_5_13
ID: 298cbr1x_15
Snippet: All RNA probes were 3' biotinylated and immobilized to a streptavidin-coated BLI probe, whereupon the binding and dissociation of SOX was measured. To prevent degradation of the probe, excess calcium ion was used in place of magnesium ( S4C-D and table S1). We also measured SOX binding to the KSHV pre-miRNA sequence used to obtain the SOX-RNA co-crystal structure (K2-31) (21). Notably, the affinity of SOX for K2-31 was even less than that of the .....
Document: All RNA probes were 3' biotinylated and immobilized to a streptavidin-coated BLI probe, whereupon the binding and dissociation of SOX was measured. To prevent degradation of the probe, excess calcium ion was used in place of magnesium ( S4C-D and table S1). We also measured SOX binding to the KSHV pre-miRNA sequence used to obtain the SOX-RNA co-crystal structure (K2-31) (21). Notably, the affinity of SOX for K2-31 was even less than that of the LIMD1 3xA-G and Zipper mutants (K d = 1.8 µM), suggesting that despite having an UGAAG motif upstream of a predicted bulge, this may not be a bona fide SOX target (Fig. 5B , S4E and table S1).
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