Assembly and activation of dynein–dynactin by the cargo adaptor protein Hook3

Metazoan cytoplasmic dynein moves processively along microtubules with the aid of dynactin and an adaptor protein that joins dynein and dynactin into a stable ternary complex. Here, we have examined how Hook3, a cargo adaptor involved in Golgi and endosome transport, forms a motile dynein-dynactin complex. We show that the conserved Hook domain interacts directly with the dynein light intermediate chain 1 (LIC1). By solving the crystal structure of the Hook domain and using structure-based mutagenesis, we identify two conserved surface residues that are each critical for LIC1 binding. Hook proteins with mutations in these residues fail to form a stable dynein-dynactin complex, revealing a crucial role for LIC1 in this interaction. We also identify a region of Hook3 specifically required for an allosteric activation of processive motility. Our work reveals the structural details of Hook3’s interaction with dynein and offers insight into how cargo adaptors form processive dynein-dynactin motor complexes.

• Publication year

  2016

• Venue

  bioRxiv

• Publication date

  2016-04-08

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1083/jcb.201604002PMID 27482052PMCID 4970328
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• How dynein and dynactin transport cargos: a structural perspective.

  2016cited by this paper
• How Dynein Moves Along Microtubules.

  2016cited by this paper
• Structural organization of the dynein-dynactin complex bound to microtubules

  2015cited by this paper
• The structure of the dynactin complex and its interaction with dynein

  2015cited by this paper
• Dynein motors: How AAA+ ring opening and closing coordinates microtubule binding and linker movement

  2015cited by this paper
• Mechanism and regulation of cytoplasmic dynein.

  2015cited by this paper
• JPred4: a protein secondary structure prediction server

  2015influential reference
• Tyrosination of α-Tubulin Controls The Initiation of Processive Dynein-Dynactin Motility

  2015cited by this paper
• FHIP and FTS proteins are critical for dynein-mediated transport of early endosomes in Aspergillus

  2014cited by this paper
• A Ras-like domain in the light intermediate chain bridges the dynein motor to a cargo-binding region

  2014influential reference
• HookA is a novel dynein–early endosome linker critical for cargo movement in vivo

  2014influential reference
• Opposing microtubule motors control motility, morphology and cargo segregation during ER-to-Golgi transport

  2014cited by this paper
• Autoinhibition and cooperative activation mechanisms of cytoplasmic dynein

  2014cited by this paper
• Hook is an adapter that coordinates kinesin-3 and dynein cargo attachment on early endosomes

  2014cited by this paper
• Activation of cytoplasmic dynein motility by dynactin-cargo adapter complexes

  2014influential reference
• In vitro reconstitution of a highly processive recombinant human dynein complex

  2014cited by this paper
• Autoregulatory Mechanism for Dynactin Control of Processive and Diffusive Dynein Transport

  2014cited by this paper
• Integrated regulation of motor-driven organelle transport by scaffolding proteins.

  2014cited by this paper
• Dlic1 deficiency impairs ciliogenesis of photoreceptors by destabilizing dynein

  2013cited by this paper
• MAFFT Multiple Sequence Alignment Software Version 7: Improvements in Performance and Usability

  2013cited by this paper
• Microtubule-dependent endosomal sorting of clathrin-independent cargo by Hook1

  2013cited by this paper
• Cytoplasmic dynein crosslinks and slides anti-parallel microtubules using its two motor domains

  2013cited by this paper
• Cytoplasmic dynein function defined by subunit composition

  2012cited by this paper
• Recruitment of dynein to late endosomes and lysosomes through light intermediate chains

  2011cited by this paper
• Cytoplasmic dynein.

  2011cited by this paper
• Mechanism and function of synaptotagmin-mediated membrane apposition

  2011cited by this paper
• Rab11-FIP3 links the Rab11 GTPase and cytoplasmic dynein to mediate transport to the endosomal-recycling compartment

  2010cited by this paper
• Computer Control of Microscopes Using µManager

  2010influential reference
• Experimental phasing with SHELXC/D/E: combining chain tracing with density modification

  2010cited by this paper
• Mitigating unwanted photophysical processes for improved single-molecule fluorescence imaging.

  2009cited by this paper
• Regulation of the processivity and intracellular localization of Saccharomyces cerevisiae dynein by dynactin

  2009cited by this paper
• Specificity of cytoplasmic dynein subunits in discrete membrane-trafficking steps.

  2009cited by this paper
• Bicaudal-D and its role in cargo sorting by microtubule-based motors.

  2009cited by this paper
• Murine CENP-F regulates centrosomal microtubule nucleation and interacts with Hook2 at the centrosome.

  2009cited by this paper
• An FTS/Hook/p107(FHIP) complex interacts with and promotes endosomal clustering by the homotypic vacuolar protein sorting complex.

  2008cited by this paper
• The Microtubule-binding Protein Hook3 Interacts with a Cytoplasmic Domain of Scavenger Receptor A*

  2007cited by this paper
• Phaser crystallographic software

  2007cited by this paper
• Iterative model building, structure refinement and density modification with the PHENIX AutoBuild wizard

  2007cited by this paper
• Hook2 Localizes to the Centrosome, Binds Directly to Centriolin/CEP110 and Contributes to Centrosomal Function

  2007cited by this paper
• Genetic Analysis of the Cytoplasmic Dynein Subunit Families

  2006cited by this paper
• Mutations in Caenorhabditis elegans Cytoplasmic Dynein Components Reveal Specificity of Neuronal Retrograde Cargo

  2004cited by this paper
• Coot: model-building tools for molecular graphics.

  2004cited by this paper
• The C. elegans hook protein, ZYG-12, mediates the essential attachment between the centrosome and nucleus.

  2003cited by this paper
• Bicaudal D induces selective dynein‐mediated microtubule minus end‐directed transport

  2003cited by this paper
• Purification of brain tubulin through two cycles of polymerization-depolymerization in a high-molarity buffer.

  2003cited by this paper
• The Golgi-Associated Hook3 Protein Is a Member of a Novel Family of Microtubule-Binding Proteins

  2001cited by this paper
• Mammalian Golgi‐associated Bicaudal‐D2 functions in the dynein–dynactin pathway by interacting with these complexes

  2001cited by this paper
• Light Intermediate Chain 1 Defines a Functional Subfraction of Cytoplasmic Dynein Which Binds to Pericentrin*

  2000cited by this paper
• Genetic analysis of hook, a gene required for endocytic trafficking in drosophila.

  1999cited by this paper
• The C . elegans Hook Protein , ZYG-12 , Mediates the Essential Attachment between the Centrosome

  1997cited by this paper
• Molecular analysis of a cytoplasmic dynein light intermediate chain reveals homology to a family of ATPases.

  1995cited by this paper
• A switch between two-, three-, and four-stranded coiled coils in GCN4 leucine zipper mutants.

  1993cited by this paper
• Predicting coiled coils from protein sequences

  1991cited by this paper
• Where is the ?c?

  1978cited by this paper
• Electronic Reprint Biological Crystallography Phenix : a Comprehensive Python-based System for Macromolecular Structure Solution

  year unknowncited by this paper
• Electronic Reprint Biological Crystallography Iterative Model Building, Structure Refinement and Density Modification with the Phenix Autobuild Wizard Biological Crystallography Iterative Model Building, Structure Refinement and Density Modification with the Phenix Autobuild Wizard

  year unknowncited by this paper

• Roles of microtubules and LIS1 in dynein transport machinery assembly.

  2026cites this paper
• Synthetic cargo adaptors reveal molecular features that can enhance dynein activation

  2025cites this paper
• Molecular basis for assembly and activation of the Hook3 − KIF1C complex-dependent transport machinery

  2025cites this paper
• Fat traffic control: S-acylation in axonal transport

  2025cites this paper
• HIV-1 binds dynein directly to hijack microtubule transport machinery

  2025influential citation
• CDR2 is a dynein adaptor recruited by kinectin to regulate ER sheet organization.

  2025cites this paper
• Regulation of angiogenesis by endocytic trafficking mediated by cytoplasmic dynein 1 light intermediate chain 1

  2024cites this paper
• Cargo specificity, regulation, and therapeutic potential of cytoplasmic dynein

  2024cites this paper
• Structure and Function of Dynein’s Non-Catalytic Subunits

  2024influential citation
• Molecular mechanism of dynein-dynactin complex assembly by LIS1

  2024influential citation
• CDR2 is a dynein adaptor recruited by kinectin to regulate ER sheet organization

  2024cites this paper
• VezA/vezatin facilitates proper assembly of the dynactin complex in vivo

  2024cites this paper
• Angiogenesis is limited by LIC1-mediated lysosomal trafficking

  2024cites this paper
• NuMA is a mitotic adaptor protein that activates dynein and connects it to microtubule minus ends

  2024cites this paper
• VezA/vezatin facilitates proper assembly of the dynactin complex in vivo

  2024cites this paper
• Structure of the endosomal Commander complex linked to Ritscher-Schinzel syndrome

  2023cites this paper
• HIV-1 binds dynein directly to hijack microtubule transport machinery

  2023cites this paper
• Kazrin promotes dynein/dynactin-dependent traffic from early to recycling endosomes

  2023cites this paper
• TRAK adaptors regulate the recruitment and activation of dynein and kinesin in mitochondrial transport

  2023cites this paper
• TRAK adaptors regulate the recruitment and activation of dynein and kinesin in mitochondrial transport

  2023cites this paper
• Structure of the Commander endosomal trafficking complex linked to X-linked intellectual disability/Ritscher-Schinzel syndrome

  2023influential citation
• High-Resolution Tracking of Dynein-Dynactin-BicD2 Complexes.

  2023cites this paper
• Conformational transitions of the mitotic adaptor Spindly underlie its interaction with Dynein and Dynactin

  2022cites this paper
• In vitro characterization of the full-length human dynein-1 cargo adaptor BicD2

  2022cites this paper
• Choreographing the motor-driven endosomal dance

  2022cites this paper
• RUFY1 binds Arl8b and mediates endosome-to-TGN CI-M6PR retrieval for cargo sorting to lysosomes

  2022cites this paper
• Conformational transitions of the Spindly adaptor underlie its interaction with Dynein and Dynactin

  2022cites this paper
• Structure of dynein–dynactin on microtubules shows tandem adaptor binding

  2022cites this paper
• Doublecortin and JIP3 are neural-specific counteracting regulators of dynein-mediated retrograde trafficking

  2022cites this paper
• Multivalency, autoinhibition, and protein disorder in the regulation of interactions of dynein intermediate chain with dynactin and the nuclear distribution protein

  2022cites this paper
• Kinesin-1 autoinhibition facilitates the initiation of dynein cargo transport

  2022cites this paper
• Selective motor activation in organelle transport along axons

  2022cites this paper
• Rab11 regulates autophagy at dendritic spines in an mTOR- and NMDA-dependent

  2022cites this paper
• Rab11 regulates autophagy at dendritic spines in an mTOR- and NMDA-dependent manner

  2022cites this paper
• The meiotic LINC complex component KASH5 is an activating adaptor for cytoplasmic dynein

  2022influential citation
• The conserved centrosomin motif, γTuNA, forms a dimer that directly activates microtubule nucleation by the γ-tubulin ring complex (γTuRC)

  2022cites this paper
• The KASH5 protein involved in meiotic chromosomal movements is a novel dynein activating adaptor

  2022cites this paper
• Dynein light intermediate chains as pivotal determinants of dynein multifunctionality.

  2021cites this paper
• Dynein activation in vivo is regulated by the nucleotide states of its AAA3 domain

  2021cites this paper
• Subcellular spatial transcriptomics identifies three mechanistically different classes of localizing RNAs

  2021cites this paper
• Phosphorylation and Pin1 binding to the LIC1 subunit selectively regulate mitotic dynein functions

  2021cites this paper
• RUSC2 and WDR47 oppositely regulate kinesin–1-dependent distribution of ATG9A to the cell periphery

  2021cites this paper
• Cryo‐EM reveals the complex architecture of dynactin's shoulder region and pointed end

  2021cites this paper
• Review: Cytoplasmic dynein motors in photoreceptors

  2021influential citation
• TRAK adaptors coordinate the recruitment and activation of dynein and kinesin to control mitochondrial transport

  2021cites this paper
• A novel BICD2 mutation of a patient with Spinal Muscular Atrophy Lower Extremity Predominant 2.

  2021influential citation
• The FTS-Hook-FHIP (FHF) complex interacts with AP-4 to mediate perinuclear distribution of AP-4 and its cargo ATG9A

  2020influential citation
• miR‐496 remedies hypoxia reoxygenation–induced H9c2 cardiomyocyte apoptosis via Hook3‐targeted PI3k/Akt/mTOR signaling pathway activation

  2020cites this paper
• Deciphering anomalous heterogeneous intracellular transport with neural networks

  2020cites this paper
• The Generation of Dynein Networks by Multi-Layered Regulation and Their Implication in Cell Division

  2020cites this paper
• IGF1R regulates retrograde axonal transport of signalling endosomes in motor neurons

  2020cites this paper
• The crosstalk between microtubules, actin and membranes shapes cell division

  2020cites this paper
• Role of dynein–dynactin complex, kinesins, motor adaptors, and their phosphorylation in dendritogenesis

  2020cites this paper
• Activation and Regulation of Cytoplasmic Dynein.

  2020influential citation
• Activation of cytoplasmic dynein through microtubule crossbridging

  2020cites this paper
• Organizational Principles of the NuMA-Dynein Interaction Interface and Implications for Mitotic Spindle Functions.

  2020cites this paper
• Stable tug-of-war between kinesin-1 and cytoplasmic dynein upon different ATP and roadblock concentrations

  2020cites this paper
• Cryo-EM reveals the complex architecture of dynactin’s shoulder and pointed end

  2020cites this paper
• Dynamic organization of intracellular organelle networks.

  2020cites this paper
• ORP1L regulates dynein clustering on endolysosmal membranes in response to cholesterol levels

  2020cites this paper
• Cargo-Mediated Activation of Cytoplasmic Dynein in vivo

  2020influential citation
• Sequential dynein effectors regulate axonal autophagosome motility in a maturation-dependent pathway

  2020cites this paper
• A tunable LIC1-adaptor interaction modulates dynein activity in a cargo-specific manner

  2020cites this paper
• Regulation of dynein activity during spindle positioning in budding yeast

  2020cites this paper
• A septin GTPase scaffold of dynein–dynactin motors triggers retrograde lysosome transport

  2020cites this paper
• Hook3 is a scaffold for the opposite-polarity microtubule-based motors cytoplasmic dynein-1 and KIF1C

  2019cites this paper
• LIS1 regulates cargo-adapter–mediated activation of dynein by overcoming its autoinhibition in vivo

  2019cites this paper
• O-cyclic phytosphingosine-1-phosphate stimulates HIF1α-dependent glycolytic reprogramming to enhance the therapeutic potential of mesenchymal stem cells

  2019cites this paper
• Insights into LIS1 function in cargo-adapter-mediated dynein activation in vivo

  2019influential citation
• Setting the dynein motor in motion: New insights from electron tomography

  2019cites this paper
• PTPN21 and Hook3 relieve KIF1C autoinhibition and activate intracellular transport

  2019cites this paper
• Incorporating Motility in the Motor: Role of the Hook Protein Family in Regulating Dynein Motility.

  2019cites this paper
• Dynein activators and adaptors at a glance

  2019cites this paper
• Role of HIF1α Regulatory Factors in Stem Cells

  2019cites this paper
• CRACR2a is a calcium-activated dynein adaptor protein that regulates endocytic traffic

  2019cites this paper
• Spatiotemporal Changes of the Phagosomal Proteome in Dendritic Cells in Response to LPS Stimulation*

  2019cites this paper
• Dynactin p150 promotes processive motility of DDB complexes by minimizing diffusional behavior of dynein

  2019cites this paper
• Hexameric NuMA:LGN structures promote multivalent interactions required for planar epithelial divisions

  2019cites this paper
• Mechanisms of Regulation of Cytoplasmic Dynein

  2019influential citation
• Investigating the role of the insulin-like growth factor receptor (IGF1R) on axonal transport as a target for pharmacological intervention in Amyotrophic Lateral Sclerosis

  2019influential citation
• A transient helix in the disordered region of dynein light intermediate chain links the motor to structurally diverse adaptors for cargo transport

  2019cites this paper
• The dynein adaptor Hook2 plays essential roles in mitotic progression and cytokinesis

  2019influential citation
• Distinct roles for dynein light intermediate chains in neurogenesis, migration, and terminal somal translocation

  2019cites this paper
• The cortical force-generating machinery: how cortical spindle-pulling forces are generated.

  2019cites this paper
• Cargo Specific Regulation of Cytoplasmic Dynein by Effector Proteins

  2018influential citation
• Spindly is required for rapid migration of human cells

  2018cites this paper
• The cytoplasmic dynein transport machinery and its many cargoes

  2018cites this paper
• A conserved interaction of the dynein light intermediate chain with dynein-dynactin effectors necessary for processivity

  2018cites this paper
• Cooperative Accumulation of Dynein-Dynactin at Microtubule Minus-Ends Drives Microtubule Network Reorganization.

  2018cites this paper
• DNAH6 is a novel candidate gene associated with sperm head anomaly

  2018cites this paper
• Rôles dans les lymphocytes T de la protéine Lis1, un régulateur de la dynamique des microtubules dépendante de la dynéine

  2018cites this paper
• Activation of intracellular transport by relieving KIF1C autoinhibition

  2018cites this paper
• Drosophila cytoplasmic dynein: Mutations, tools, and developmental functions

  2018cites this paper
• Regulation of cytoplasmic dynein motility

  2018cites this paper
• A new calcium-activated dynein adaptor protein, CRACR2a, regulates clathrin-independent endocytic traffic in T cells

  2018cites this paper
• Multiple Roles, Multiple Adaptors: Dynein During Cell Cycle.

  2018cites this paper
• Biochemical and Biophysical Roles of Cell Surface Molecules

  2018cites this paper
• CLIP-170 is essential for MTOC repositioning during T cell activation by regulating dynein localisation on the cell surface

  2018cites this paper
• Dynein activator Hook1 is required for trafficking of BDNF-signaling endosomes in neurons

  2018influential citation
• Molecular mechanisms of the interhead coordination by interhead tension in cytoplasmic dyneins

  2018cites this paper