Cytoplasmic dynein is the predominant minus-end-directed microtubule (MT) motor in most eukaryotic cells. In addition to transporting vesicular cargos, dynein helps to organize MTs within MT networks such as mitotic spindles. How dynein performs such non-canonical functions is unknown. Here we demonstrate that dynein crosslinks and slides anti-parallel MTs in vitro. Surprisingly, a minimal dimeric motor lacking a tail domain and associated subunits can cause MT sliding. Single molecule imaging reveals that motors pause and frequently reverse direction when encountering an anti-parallel MT overlap, suggesting that the two motor domains can bind both MTs simultaneously. In the mitotic spindle, inward microtubule sliding by dynein counteracts outward sliding generated by kinesin-5, and we show that a tailless, dimeric motor is sufficient to drive this activity in mammalian cells. Our results identify an unexpected mechanism for dynein-driven microtubule sliding, which differs from filament sliding mechanisms described for other motor proteins.
When cells divide, they must also divide their contents. In particular, both ‘mother’ and ‘daughter’ cells require full sets of chromosomes, which must first be duplicated, and then evenly distributed between the cells. Protein filaments called microtubules form a network that helps to accurately segregate the chromosomes. Microtubules emanate from structures at each end of the dividing cell known as spindle poles; after the chromosomes have duplicated, the microtubules latch onto them and align the pairs in the middle of the cell. As the two cells separate, microtubules at opposite spindle poles reel in one chromosome from each pair.
Microtubules are composed of alternating copies of two different types of a protein called tubulin, and have ends with distinct properties. The ‘minus’ ends are directed outwards, away from the chromosomes; the ‘plus’ ends—which can actively add tubulin—grow toward the middle of the cell, and can also bind to chromosomes. Microtubules can be manipulated by motor proteins that ‘walk’ along them carrying cargoes, which can include other microtubules. The combined actions of many motor proteins rearrange the microtubule network into a configuration that enables the chromosomes, and other cellular structures, to partition equally between the mother and daughter cells.
Motor proteins such as dynein and kinesin transport cargoes along microtubules; each motor is composed of two identical copies of the protein bound to each other. Kinesin walks toward the plus end of a microtubule, propelling itself using ‘feet’ that are called motor domains; it binds cargoes (including other microtubules) through additional regions located at the opposite end of the protein. In contrast, dynein walks toward the minus end of a microtubule. Although dynein is known to carry certain cargoes through regions outside its motor domain, how it transports other microtubules is not well understood.
Tanenbaum et al. now show that regions outside the motor domain of dynein are unnecessary to transport microtubule cargoes. When two dynein motor domains are isolated and linked to each other in vitro, each can bind to a separate microtubule. By walking toward the minus ends of their respective microtubules, the motor domains drive the microtubules in opposite directions, sliding them apart. These studies thus provide insight into the mechanism through which dynein works with additional motor proteins (such as kinesin) to rearrange microtubules during cell division—and also to ensure that chromosomes segregate evenly between mother and daughter cells.