Dynein activation in vivo is regulated by the nucleotide states of its AAA3 domain

Qiu, Rongde; Zhang, Jun; Rotty, Jeremy D.; Xiang, Xin

doi:10.1101/2021.04.12.439451

Cited by 5 publications

(5 citation statements)

References 156 publications

(326 reference statements)

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“…A similar pathway for dynein regulation may exist in other organisms, including mammalian cells (Figure 8B), and related mechanisms have recently been proposed (Markus et al, 2020;Qiu et al, 2021). Our work shows that both the sitering and sitestalk Lis1 binding sites on dynein are important for the formation of active human dyneindynactin-activating adaptor complexes containing either the Hook3 or BicD2 activating adaptors.…”

Section: Discussionsupporting

confidence: 81%

Structural Basis for Cytoplasmic Dynein-1 Regulation by Lis1

Reimer

Karasmanis

et al. 2021

Preprint

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The lissencephaly 1 gene, LIS1, is mutated in patients with the neurodevelopmental disease lissencephaly. The Lis1 protein is conserved from fungi to mammals and is a key regulator of cytoplasmic dynein-1, the major minus-end-directed microtubule motor in many eukaryotes. Lis1 is the only dynein regulator that binds directly to dynein's motor domain, and by doing so alters dynein's mechanochemistry. Lis1 is required for the formation of fully active dynein complexes, which also contain essential cofactors: dynactin and an activating adaptor. Here, we report the first high-resolution structure of the yeast dynein-Lis1 complex. Our 3.1Å structure reveals, in molecular detail, the major contacts between dynein and Lis1 and between Lis1's β-propellers. Structure-guided mutations in Lis1 and dynein show that these contacts are required for Lis1's ability to form fully active human dynein complexes and to regulate yeast dynein's mechanochemistry and in vivo function. We present a model for the conserved role of Lis1 in regulating dynein from yeast to humans.

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Section: Discussionsupporting

confidence: 81%

Structural Basis for Cytoplasmic Dynein-1 Regulation by Lis1

Reimer

Karasmanis

et al. 2021

Preprint

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“…While dynein's microtubule binding domain is not required to reach the cortex (Lammers and Markus, 2015), our data suggest some role for microtubule binding in this process. Assembling active dynein-dynactinactivating adaptor complexes likely involves multiple steps and the many in vitro phenotypes described for Lis1's regulation of dynein, such as the nucleotide-specific affects at AAA3 (DeSantis et al, 2017;Qiu et al, 2021), may be providing mechanistic hints about these steps.…”

Section: Discussionmentioning

confidence: 99%

“…Genetically, mutations that link nucleotide hydrolysis at AAA3 to Lis1 have also been reported (Qiu et al, 2021;Willins et al, 1997;Zhuang et al, 2007). While it remains to be determined how the readouts described above or other in vitro readouts of Lis1 regulation of dynein (Baumbach et al, 2017;Gutierrez et al, 2017;Jha et al, 2017;Marzo et al, 2020;Wang et al, 2013) relate to Lis1's in vivo function, they serve as powerful tools to test structure/ function predictions (DeSantis et al, 2017;Toropova et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

Structural basis for cytoplasmic dynein-1 regulation by Lis1

Gillies

Reimer

Karasmanis

et al. 2022

eLife

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The lissencephaly 1 gene, LIS1, is mutated in patients with the neurodevelopmental disease lissencephaly. The Lis1 protein is conserved from fungi to mammals and is a key regulator of cytoplasmic dynein-1, the major minus-end-directed microtubule motor in many eukaryotes. Lis1 is the only dynein regulator known to bind directly to dynein's motor domain, and by doing so alters dynein's mechanochemistry. Lis1 is required for the formation of fully active dynein complexes, which also contain essential cofactors: dynactin and an activating adaptor. Here, we report the first high-resolution structure of the yeast dynein–Lis1 complex. Our 3.1Å structure reveals, in molecular detail, the major contacts between dynein and Lis1 and between Lis1's ß-propellers. Structure-guided mutations in Lis1 and dynein show that these contacts are required for Lis1's ability to form fully active human dynein complexes and to regulate yeast dynein's mechanochemistry and in vivo function.

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“…Studies indicate that AAA1 is the main site for ATPase activity 42,45,47 , with AAA3 and AAA4 playing regulatory roles during motility and force production [48][49][50][51] . Previous studies have also found that the nucleotide state of AAA3 may play a direct role in modulating dynein-Pac1 affinity 52,53 . To determine whether ATP binding or hydrolysis at AAA1, 3 or 4 might affect the ability of dyneinMOTOR MT-B or dyneinMOTOR MT-U , we mutated either Walker A or Walker B ("WA", or "WB"; responsible for ATP binding and hydrolysis, respectively) in AAA1, 3 and 4, and assessed the extent of their plus end binding.…”

Section: The Nucleotide-bound State Of Dynein Affects Its Affinity Fo...mentioning

confidence: 96%

Microtubule binding-induced allostery promotes LIS1 dissociation from dynein prior to cargo transport

Ton

Wang

Chai

et al. 2022

Preprint

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The lissencephaly-related protein LIS1 is a critical regulator of cytoplasmic dynein that governs motor function and intracellular localization (e.g., 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 -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 bound to microtubule plus-ends. We find that a monomeric motor domain is sufficient to exhibit these opposing LIS1 affinities, and that this is an evolutionarily conserved phenomenon. Three cryo-EM structures of 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.

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Dynein activation in vivo is regulated by the nucleotide states of its AAA3 domain

Cited by 5 publications

References 156 publications

Structural Basis for Cytoplasmic Dynein-1 Regulation by Lis1

Structural Basis for Cytoplasmic Dynein-1 Regulation by Lis1

Structural basis for cytoplasmic dynein-1 regulation by Lis1

Microtubule binding-induced allostery promotes LIS1 dissociation from dynein prior to cargo transport

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