Structure of a Yeast Dyn2-Nup159 Complex and Molecular Basis for Dynein Light Chain-Nuclear Pore Interaction

The Mechanism of Dynein Light Chain LC8-mediated Oligomerization of the Ana2 Centriole Duplication Factor

“…As reported previously (29), purified LC8 eluted primarily as a dimer (Fig. 6A, light green trace indicating the Rayleigh ratio) with a mass of 21.6 kDa (Fig.…”

“…Protein Purification for Crystallization-LC8 was purified as described previously for the yeast homologue Dyn2 (29). Briefly, cells expressing GST-LC8 were lysed by sonication and clarified by centrifugation at 23,000 ϫ g for 45 min, and the supernatant was loaded onto a Glutathione Sepharose column (GE Healthcare).…”

Multiple Recognition Motifs in Nucleoporin Nup159 Provide a Stable and Rigid Nup159-Dyn2 Assembly

“…tures of bound LC8 complexes to date show partner proteins adopting a sixth ␤-strand in an LC8 antiparallel ␤-sheet (6,7,13,16,17), and the Nup-Dyn2 complexes are expected to have the same (6, 7); thus the QT3 motif containing a segment with helical propensity is less likely to favor acquisition of a ␤-strand conformation at the binding interface with Dyn2.…”

“…In studies of the assembly of Nup159-Nup82-Nsp1 nucleoporin subcomplex, Hurt and co-workers (5) identified Dyn2, the yeast ortholog of LC8, as a binding partner of Nup159. Dyn2 and Drosophila melanogaster LC8 are very closely related, with 48% sequence identity and essentially the same three-dimensional structure (6,7). LC8, a dynein light chain that also binds numerous other non-dynein partners, has been identified as a versatile hub protein that in diverse systems binds to disordered protein segments and assists folding, assembly, and regulation of multiprotein complexes (8).…”

“…This binding enhancement involving multiple bivalent dimers is termed "polybivalency" (9, 10) and the IC-light chain assemblage is a canonical example of polybivalency in intrinsically disordered proteins. A common feature among LC8 binding partners, including dynein IC, is that the LC8 recognition motif is disordered in the apo form but is ordered in the LC8-bound form where it packs as a ␤-strand that spans the length of LC8 dimer interface (6,(13)(14)(15)(16)(17).…”