Lis1 regulates dynein by sterically blocking its mechanochemical cycle.
Lis1 regulates dynein by sterically blocking its mechanochemical cycle.
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DOI：
10.7554/elife.03372
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发表时间：
2014-11-07
期刊：
影响因子：
7.7
通讯作者：
Leschziner AE
中科院分区：
文献类型：
--
作者：
Toropova K;Zou S;Roberts AJ;Redwine WB;Goodman BS;Reck-Peterson SL;Leschziner AE
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 Cells use motor proteins to move ‘cargo’ from one location to another inside the cell. This cargo can range in size from a single macromolecule to something as large as the nucleus of the cell. A motor protein called dynein is the largest and least understood of the motor proteins found in cells. Dynein molecules work in pairs to take ‘steps’ along tracks called microtubules. Dynein contains two domains: a motor domain, which is responsible for generating movement, and a ‘tail’ domain to which the cargo is attached. The motor domain is composed of a ring-like shape and two appendages—the stalk and the linker. The linker undergoes large-scale movements relative to the ring that transmits force to the tail domain. Dynein also interacts with various accessory proteins to do its job inside the cell. One of these is a protein called Lis1 that is found across a wide range of species from yeast to humans. Defects in the gene for Lis1 result in brain developmental disorders in humans. However, it is not clear how the Lis1 protein influences the activity of dynein. Now Toropova, Zou et al. have visualized the structure of dynein bound to Lis1 and compared it with the structure of dynein on its own in order to work out if dynein changes its shape as a result of binding to Lis1. These experiments show that when Lis1 binds to dynein, it physically blocks the linker, preventing it from making contacts with the ring-like shape that are important for the normal function of the motor. To test the idea that this physical block is responsible for dynein molecules spending a relatively long time attached to their microtubules, Toropova, Zou et al. shortened the linker to a point where the Lis1 protein could no longer block it: this resulted in a dynein motor that was no longer sensitive to Lis1. A challenge for the future is to understand, at a molecular level, how the Lis1-mediated slowing down of dynein affects the multiple functions the motor carries out in a cell. DOI: http://dx.doi.org/10.7554/eLife.03372.002
