| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | A synthetic gene encoding human RCC1 (Supplementary Table 3) was synthesized by GenScript, while yeast RCC1 was PCR out from a Saccharomyces cerevisiae (strain S288C) genomic library....n. Clarified lysates expressing GST-tagged proteins were incubated with glutathione resin (GenScript) and beads were washed with Complex buffer...A volume of 700 μl of this mixture was then incubated with 100 µl of glutathione resin beads (GenScript) ... | Get A Quote |
| Proteins, Expression, Isolation and Analysis | Get A Quote |
Active nuclear import of Ran exchange factor RCC1 is mediated by importin α3. This pathway is essential to generate a gradient of RanGTP on chromatin that directs nucleocytoplasmic transport, mitotic spindle assembly and nuclear envelope formation. Here we identify the mechanisms of importin α3 selectivity for RCC1. We find this isoform binds RCC1 with one order of magnitude higher affinity than the generic importin α1, although the two isoforms share an identical NLS-binding groove. Importin α3 uses its greater conformational flexibility to wedge the RCC1 β-propeller flanking the NLS against its lateral surface, preventing steric clashes with its Armadillo-core. Removing the β-propeller, or inserting a l... More
