Cytoplasmic dynein is a microtubule-based motor required for intracellular transport and cell division. Its movement involves coupling cycles of track binding and release with cycles of force-generating nucleotide hydrolysis. How this is accomplished given the ~25 nm separating dynein’s track- and nucleotide-binding sites is not understood. Here, we present a sub-nanometer-resolution structure of dynein’s microtubule-binding domain bound to microtubules by cryo-electron microscopy that was used to generate a pseudo-atomic model of the complex with molecular dynamics. We identified large rearrangements triggered by track binding and specific interactions, confirmed by mutagenesis and single molecule motility assays, which tune dynein’s affinity for microtubules. Our results provide a molecular model for how dynein’s binding to microtubules is communicated to the rest of the motor.

Regulation of cytoplasmic dynein's motor activity is essential for diverse eukaryotic functions, including cell division, intracellular transport, and brain development. The dynein regulator Lis1 is known to keep dynein bound to microtubules; however, how this is accomplished mechanistically remains unknown. We have used three-dimensional electron microscopy, single-molecule imaging, biochemistry, and in vivo assays to help establish this mechanism. The three-dimensional structure of the dynein–Lis1 complex shows that binding of Lis1 to dynein's AAA+ ring sterically prevents dynein's main mechanical element, the ‘linker’, from completing its normal conformational cycle. Single-molecule experiments show that eliminating this block by shortening the linker to a point where it can physically bypass Lis1 renders single dynein motors insensitive to regulation by Lis1. Our data reveal that Lis1 keeps dynein in a persistent microtubule-bound state by directly blocking the progression of its mechanochemical cycle.DOI: http://dx.doi.org/10.7554/eLife.03372.001

In the present study, we have investigated the growth of a multilayer formed by poly(sodium 4-styrene sulfonate) (PSSS) and poly(diallyldimethylammonium chloride) (PDDA) at different salt concentrations by use of quartz crystal microbalance with dissipation (QCM-D). The frequency change (Deltaf) demonstrates that the exponential growth mode gradually becomes dominant as NaCl concentration (C(NaCl)) increases. On the other hand, the dissipation change (DeltaD) reveals that the deposition is dominated by chain conformation at C(NaCl) < 1.0 M, where the change of the characteristic growth parameter agrees well with the results fit with Debye length. At C(NaCl) > or = 1.0 M, the growth is not determined by chain conformation but by chain interpenetration.