8FDU

Engineered human dynein motor domain in the microtubule-unbound state with LIS1 complex in the buffer containing ATP-Vi (local refined on AAA3-AAA5 and LIS1)

Summary for 8FDU

• Entry DOI: 10.2210/pdb8fdu/pdb
• EMDB information: 29012 29014
• Descriptor: Cytoplasmic dynein 1 heavy chain 1,Serine--tRNA ligase, Platelet-activating factor acetylhydrolase IB subunit beta,Platelet-activating factor acetylhydrolase IB subunit beta,human LIS1 protein with a SNAP tag, ADENOSINE-5'-DIPHOSPHATE (3 entities in total)
• Functional Keywords: dynein, motor domain, microtubule-unbound, motor protein, lis1
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 3
• Total formula weight: 491160.62

Authors

Ton, W.,Wang, Y.,Chai, P. (deposition date: 2022-12-04, release date: 2023-06-21, Last modification date: 2023-11-15)

Primary citation

Ton, W.D.,Wang, Y.,Chai, P.,Beauchamp-Perez, C.,Flint, N.T.,Lammers, L.G.,Xiong, H.,Zhang, K.,Markus, S.M.
Microtubule-binding-induced allostery triggers LIS1 dissociation from dynein prior to cargo transport.
Nat.Struct.Mol.Biol., 30:1365-1379, 2023

PubMed Abstract: The lissencephaly-related protein LIS1 is a critical regulator of cytoplasmic dynein that governs motor function and intracellular localization (for example, to microtubule plus-ends). Although LIS1 binding is required for dynein activity, its unbinding prior to initiation of cargo transport is equally important, since preventing dissociation leads to dynein dysfunction. To understand whether and how dynein-LIS1 binding is modulated, we engineered dynein mutants locked in a microtubule-bound (MT-B) or microtubule-unbound (MT-U) state. Whereas the MT-B mutant exhibits low LIS1 affinity, the MT-U mutant binds LIS1 with high affinity, and as a consequence remains almost irreversibly associated with microtubule plus-ends. We find that a monomeric motor domain is sufficient to exhibit these opposing LIS1 affinities, and that this is evolutionarily conserved between yeast and humans. Three cryo-EM structures of human dynein with and without LIS1 reveal microtubule-binding induced conformational changes responsible for this regulation. Our work reveals key biochemical and structural insight into LIS1-mediated dynein activation.

PubMed: 37322240
DOI: 10.1038/s41594-023-01010-x

Experimental method

ELECTRON MICROSCOPY (3.3 Å)