Lis1 Acts as a “Clutch” between the ATPase and Microtubule-Binding Domains of the Dynein Motor

Huang, Jinxiu; Roberts, A. J.; Leschziner, Andres E.; Reck-Peterson, Samara L.

doi:10.1016/j.cell.2012.07.022

Cited by 218 publications

(440 citation statements)

References 67 publications

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“…Although both regulators are known to interact with both dynein and dynactin, currently no overlapping binding sites are known. Lis1 binds the AAA+ ring of the dynein motor domain (Tai et al , 2002; Huang et al , 2012) and the p50 subunit of dynactin (Tai et al , 2002), whereas BicD2 interacts with the dynein tail and the Arp1 filament of dynactin (Chowdhury et al , 2015; Urnavicius et al , 2015). Interestingly, Lis1 has been shown to bind to a compact dynein conformation with docked motor domains (also known as the phi particle) (Toba et al , 2015) that has been proposed to correspond to an autoinhibited state of the dynein motor (Torisawa et al , 2014).…”

Section: Discussionmentioning

confidence: 99%

Combinatorial regulation of the balance between dynein microtubule end accumulation and initiation of directed motility

et al. 2017

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Cytoplasmic dynein is involved in a multitude of essential cellular functions. Dynein's activity is controlled by the combinatorial action of several regulatory proteins. The molecular mechanism of this regulation is still poorly understood. Using purified proteins, we reconstitute the regulation of the human dynein complex by three prominent regulators on dynamic microtubules in the presence of end binding proteins (EBs). We find that dynein can be in biochemically and functionally distinct pools: either tracking dynamic microtubule plus‐ends in an EB‐dependent manner or moving processively towards minus ends in an adaptor protein‐dependent manner. Whereas both dynein pools share the dynactin complex, they have opposite preferences for binding other regulators, either the adaptor protein Bicaudal‐D2 (BicD2) or the multifunctional regulator Lissencephaly‐1 (Lis1). BicD2 and Lis1 together control the overall efficiency of motility initiation. Remarkably, dynactin can bias motility initiation locally from microtubule plus ends by autonomous plus‐end recognition. This bias is further enhanced by EBs and Lis1. Our study provides insight into the mechanism of dynein regulation by dissecting the distinct functional contributions of the individual members of a dynein regulatory network.

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Section: Discussionmentioning

confidence: 99%

Combinatorial regulation of the balance between dynein microtubule end accumulation and initiation of directed motility

et al. 2017

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“…In the absence of load, ATP at AAA3 blocks reorientation of the linker from the post-to the prepowerstroke conformation (9). In addition, binding of the cofactor Lis1, which mechanically obstructs linker movements, uncouples AAA1's ATPase activities from changes in MT-binding affinity (44). Finally, AAA5 mutations preventing linker docking severely reduce dynein's ATPase activities (3,8).…”

Section: Discussionmentioning

confidence: 99%

Cytoplasmic dynein regulates its attachment to microtubules via nucleotide state-switched mechanosensing at multiple AAA domains

Nicholas

Berger

Rao³

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

115

212

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Cytoplasmic dynein is a homodimeric microtubule (MT) motor protein responsible for most MT minus-end-directed motility. Dynein contains four AAA+ ATPases (AAA: ATPase associated with various cellular activities) per motor domain (AAA1-4). The main site of ATP hydrolysis, AAA1, is the only site considered by most dynein motility models. However, it remains unclear how ATPase activity and MT binding are coordinated within and between dynein's motor domains. Using optical tweezers, we characterize the MT-binding strength of recombinant dynein monomers as a function of mechanical tension and nucleotide state. Dynein responds anisotropically to tension, binding tighter to MTs when pulled toward the MT plus end. We provide evidence that this behavior results from an asymmetrical bond that acts as a slip bond under forward tension and a slip-ideal bond under backward tension. ATP weakens MT binding and reduces bond strength anisotropy, and unexpectedly, so does ADP. Using nucleotide binding and hydrolysis mutants, we show that, although ATP exerts its effects via binding AAA1, ADP effects are mediated by AAA3. Finally, we demonstrate "gating" of AAA1 function by AAA3. When tension is absent or applied via dynein's C terminus, ATP binding to AAA1 induces MT release only if AAA3 is in the posthydrolysis state. However, when tension is applied to the linker, ATP binding to AAA3 is sufficient to "open" the gate. These results elucidate the mechanisms of dynein-MT interactions, identify regulatory roles for AAA3, and help define the interplay between mechanical tension and nucleotide state in regulating dynein motility.cytoplasmic dynein | mechanosensing | optical tweezers | AAA+ ATPases | microtubules N umerous eukaryotic cellular processes require motion and force generated by cytoskeletal motor proteins, among which cytoplasmic dynein (hereinafter, "dynein") is unique for its size, complexity, and versatility. As a homodimeric, divergent AAA+ ATPase (AAA: ATPase associated with various cellular activities), dynein drives the majority of microtubule (MT) minusend-directed motility in most eukaryotes (1). The motor functions as a massive protein complex (2), but its catalytic core consists of two identical heavy chains, each with six AAA modules (AAA1-6) linked in tandem to form a ring (Fig. 1A). AAA1-4 bind nucleotides, whereas AAA5 and -6 are structural (3, 4). A ∼15-nm "stalk" emerging from AAA4 (3, 4) separates the AAA modules from the MT-binding domain (MTBD). The stalk configuration influences both MT affinity and ATPase activity (5) and thereby mediates bidirectional allosteric communication between the AAA ring and the MTBD (3, 6). Finally, a ∼10-nm "linker" also emerges from the ring and undergoes cyclic reorientations that generate force and displacement (7-9).For dynein to "walk," one motor domain ("head") must remain MT-bound while the other moves (10-13), thus requiring coordination of the "internal" cycles of both heads. Dynein may use allosteric mechanosensing (possibly through the stalk) to differentiate be...

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“…Rab6a(Q72L) has significantly higher affinity for the LIS1-dynein complex, suggesting that the molecular mechanism of Rab6a(Q72L)-mediated LIS1 dissociation might not be attributable to simple competition. The LIS1-binding position on dynein was recently determined 32 . Rab6a might induce a structural modification, resulting in decreased LIS1-binding affinity.…”

Section: Discussionmentioning

confidence: 99%

“…S4e,f, Supplementary Movie 10). Although our in vitro singlemolecule assays indicate that LIS1 inhibits dynein motility, it might be possible that LIS1-bound dynein is still mobile within the small level, in which the spatial resolution of our method cannot dissect 32 . Using this system, we examined whether Rab6a is able to release LIS1 from the LIS1-dynein complex and to activate dynein motility.…”

Section: Rab6a(q72l) Releases Lis1 and Activates Dynein Motilitymentioning

confidence: 99%

Rab6a releases LIS1 from a dynein idling complex and activates dynein for retrograde movement

et al. 2013

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Cytoplasmic dynein drives the movement of a wide range of cargoes towards the minus ends of microtubules. We previously demonstrated that LIS1 forms an idling complex with dynein, which is transported to the plus ends of microtubules by kinesin motors. Here we report that the small GTPase Rab6a is essential for activation of idling dynein. Immunoprecipitation and microtubule pull-down assays reveal that the GTP bound mutant, Rab6a(Q72L), dissociates LIS1 from a LIS1-dynein complex, activating dynein movement in in vitro microtubule gliding assays. We monitor transient interaction between Rab6a(Q72L) and dynein in vivo using dual-colour fluorescence cross-correlation spectroscopy in dorsal root ganglion (DRG) neurons. Finally, we demonstrate that Rab6a(Q72L) mediates LIS1 release from a LIS1-dynein complex followed by dynein activation through an in vitro single-molecule assay using triplecolour quantum dots. Our findings reveal a surprising function for GTP bound Rab6a as an activator of idling dynein.

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Lis1 Acts as a “Clutch” between the ATPase and Microtubule-Binding Domains of the Dynein Motor

Cited by 218 publications

References 67 publications

Combinatorial regulation of the balance between dynein microtubule end accumulation and initiation of directed motility

Combinatorial regulation of the balance between dynein microtubule end accumulation and initiation of directed motility

Cytoplasmic dynein regulates its attachment to microtubules via nucleotide state-switched mechanosensing at multiple AAA domains

Rab6a releases LIS1 from a dynein idling complex and activates dynein for retrograde movement

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