Cryo-EM shows how dynactin recruits two dyneins for faster movement

Urnavicius, L.; Lau, C.K.; Elshenawy, Mohamed M.; Morales-Ríos, Edgar; Motz, C.; Yıldız, Ahmet; Carter, Andrew P.

doi:10.1038/nature25462

Cited by 273 publications

(440 citation statements)

References 56 publications

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“…In one conformation, the alpha helix extends out from the body of the mushroom; in the other, it is oriented closer toward the surface of the α-subunit’s globule domain (PDB 3AAE, 11ZN,2KZ7, 2KXP) (Takeda et al, 2010; Yamashita et al, 2003; Zwolak et al, 2010a). Structures of CP bound to the actin-like Arp1 filament in dynactin complex, derived from cryoEM, also show the β-tentacle in an α-helical conformation, in contact with actin (Urnavicius et al, 2015; Urnavicius et al, 2018), and in a position consistent with the model for CP binding to actin (Narita et al, 2006). …”

Section: Resultsmentioning

confidence: 55%

Allosteric Coupling of CARMIL and V-1 Binding to Capping Protein Revealed by Hydrogen-Deuterium Exchange

et al. 2018

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Summary Actin assembly is important for cell motility. The ability of actin subunits to join or leave filaments via the barbed end is critical to actin dynamics. Capping protein (CP) binds to barbed ends to prevent subunit gain and loss, and is regulated by proteins that include V-1 and CARMIL. V-1 inhibits CP by sterically blocking one binding site for actin. CARMILs bind at a distal site and decrease the affinity of CP for actin, suggested to be caused by conformational changes. We used hydrogen-deuterium exchange with mass spectrometry (HDX-MS) to probe changes in structural dynamics induced by V-1 and CARMIL binding to CP. V-1 and CARMIL induce changes in both proteins’ binding sites on the surface of CP, along with a set of internal residues. Both also affect the conformation of CP’s β-subunit “tentacle,” a second distal actin-binding site. Concerted regulation of actin assembly by CP occurs through allosteric couplings between CP modulator and actin binding sites.

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Section: Resultsmentioning

confidence: 55%

Allosteric Coupling of CARMIL and V-1 Binding to Capping Protein Revealed by Hydrogen-Deuterium Exchange

et al. 2018

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“…Structural analysis revealed that most of these mutations (7 out of the 8) clustered to two distinct regions: (1) adjacent to, or within the N-terminal dimerization domain, or (2) at a surface that interfaces with the intermediate chain of a neighboring heavy chain in a 2 dynein:1 dynactin complex ( Figure 2, Supplement 4). This latter region was recently identified as being important to stabilize the binding of a second dynein complex to dynactin, and ensuring that all four heavy chains are properly aligned for efficient motility of the human dynein-dynactin complex 74 . Our data suggest that the ability to recruit 2 dynein complexes to dynactin is conserved in yeast, and that disrupting this complex can compromise force generation ( Fig.…”

Section: Spindle Tracking In Live Cells Provides a Sensitive Read Outmentioning

confidence: 99%

Molecular basis for dyneinopathies reveals insight into dynein regulation and dysfunction

Marzo

Griswold

Ruff

et al. 2019

Preprint

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Cytoplasmic dynein plays critical roles within the developing and mature nervous systems, including effecting nuclear migration, and retrograde transport of various cargos. Unsurprisingly, mutations in dynein are causative of various developmental neuropathies and motor neuron diseases. These "dyneinopathies" define a broad spectrum of diseases with no known correlation between mutation identity and disease state. To overcome complications associated with studying dynein function in human cells, we employed budding yeast as a screening platform to characterize the motility properties of seventeen disease-correlated dynein mutants. Using this system, we have determined the molecular basis for several broad classes of etiologically related diseases. Moreover, by engineering compensatory mutations, we have alleviated the mutant phenotypes in two of these cases, one of which we confirmed with recombinant human dynein complexes. In addition to revealing molecular insight into dynein regulation, our data reveal an unexpected correlation between the degree of dynein dysfunction and disease type.3

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“…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).…”

Section: Which Molecular Mechanisms Could Regulate These Forces?mentioning

confidence: 99%

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

Bouvrais

Chesneau

Cunff

et al. 2019

Preprint

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In the Caenorhabditis elegans zygote, astral microtubules generate forces, pushing against and pulling from the cell periphery. They are essential to position the mitotic spindle and in turn the cytokinesis furrow, ensuring the proper distribution of fate determinants to the daughter cells. By measuring the dynamics of astral microtubules, we revealed the presence of two populations, residing at the cortex during 0.4 s and 1.8 s, proposed to reflect the pulling and pushing events, respectively. This is a unique opportunity to unravel the time and space variations of these both spindle-positioning forces, to study their regulation under physiological conditions. By an investigation at the microscopic level, we first confirmed that the asymmetry in pulling forces was encoded by an anteroposterior imbalance in dynein-engaging rate, and that this asymmetry exists from early metaphase on and accounts for the final spindle position. More importantly, we obtained direct proof of the temporal control of pulling forces through the forcegenerator processivity increase during anaphase. Lastly, we discovered an anti-correlation between the long-lived population density and the stability of the spindle position during metaphase, which strongly suggests that the pushing forces contribute to maintaining the spindle at the cell centre.

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Cryo-EM shows how dynactin recruits two dyneins for faster movement

Cited by 273 publications

References 56 publications

Allosteric Coupling of CARMIL and V-1 Binding to Capping Protein Revealed by Hydrogen-Deuterium Exchange

Allosteric Coupling of CARMIL and V-1 Binding to Capping Protein Revealed by Hydrogen-Deuterium Exchange

Molecular basis for dyneinopathies reveals insight into dynein regulation and dysfunction

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

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