Lis1 Has Two Opposing Modes of Regulating Cytoplasmic Dynein

Abstract

Summary Regulation is central to the functional versatility of cytoplasmic dynein, a motor involved in intracellular transport, cell division, and neurodevelopment. Previous work established that Lis1, a conserved regulator of dynein, binds to its motor domain and induces a tight microtubule-binding state in dynein. The work we present here—a combination of biochemistry, single-molecule assays, and cryo-electron microscopy—led to the surprising discovery that Lis1 has two opposing modes of regulating dynein, being capable of inducing both low and high affinity for the microtubule. We show that these opposing modes depend on the stoichiometry of Lis1 binding to dynein and that this stoichiometry is regulated by the nucleotide state of dynein’s AAA3 domain. The low affinity state requires Lis1 binding to dynein at a novel conserved site, mutation of which disrupts Lis1’s function in vivo. We propose a new model for the regulation of dynein by Lis1.

CENP-F controls force generation and the dynein-dependent stripping of CENP-E at kinetochores

“…This context-dependent regulation explains how perturbation of Lis1 in cells can both augment and reduce cargo transport at distinct subcellular locations (Dix et al, 2013;Klinman and Holzbaur, 2015;Moughamian et al, 2013;Pandey and Smith, 2011;Shao et al, 2013;Vagnoni et al, 2016;Yi et al, 2011). Our data supports the idea that CENP-F/Ndel1/Nde1/Lis1 at human kinetochores favours a highbinding low-velocity dynein motor state that attenuates CENP-E stripping after end-on attachment (DeSantis et al, 2017). This idea will need testing with in vitro reconstitution experiments.…”

“…Lis1 is a highly conserved and wellestablished dynein regulator that is mutated in the neurodevelopmental disease Type-1 lissencephaly (Reiner and Coquelle, 2005). Lis1 binds directly to the dynein motor domain and differentially regulates its transport in response to AAA3 nucleotide state (DeSantis et al, 2017;Huang et al, 2012;Toropova et al, 2014). This context-dependent regulation explains how perturbation of Lis1 in cells can both augment and reduce cargo transport at distinct subcellular locations (Dix et al, 2013;Klinman and Holzbaur, 2015;Moughamian et al, 2013;Pandey and Smith, 2011;Shao et al, 2013;Vagnoni et al, 2016;Yi et al, 2011).…”

The coordination of spindle-positioning forces during the asymmetric division of theC. eleganszygote is revealed by distinct microtubule dynamics at the cortex

“…The first and obvious possible player is dynactin, known to be essential to cortical pulling forces and able to make dynein more processive (Skop, 1998;Gonczy et al, 1999;King and Schroer, 2000;Rodriguez-Garcia et al, 2018). Beyond, some other players were involved in other systems in vitro or in vivo, for example: the end-binding (EB) proteins putatively involved in initiating dynein pulling once a microtubule is captured (Jha et al, 2017), and whose homolog EBP-2 contributes, although modestly, to cortical forces (Rodriguez-Garcia et al, 2018); LIS-1 essential for spindle positioning in nematode and reported to be also implicated in dynein regulation beyond its classic inhibitory role (Cockell et al, 2004;Baumbach et al, 2017;DeSantis et al, 2017); BICD-1 BICD2 involved in nuclear migration in nematode hypodermis (Fridolfsson et al, 2010) but no strong early embryonic phenotype was reported, despite indication of its role in other organisms (Swan et al, 1999;Splinter et al, 2012;Jha et al, 2017;Urnavicius et al, 2018). Thirdly and finally, efficient pushing by microtubules against the cortex requires some microtubuleassociated proteins to prevent the switch to catastrophe (Janson et al, 2003).…”

Lis1 regulates Phagocytosis by co-localising with actin in the Phagocytic cup

“…Actin is subject to regulation by a host of actin associated proteins including nucleating, filament severing, end-capping, bundling, membrane tethering, and scaffolding proteins. Lissencephaly-1 (Lis1) is a protein that is classically known to interact with, and modulate the function of microtubule-based dynein motors [2] [3]. Mutations in LIS1 cause severe developmental defects in formation of the brain (Lissencephaly).…”