1. Dynamic multimerization of Dab2-Myosin VI complexes regulates cargo processivity while minimizing cortical actin reorganization.
- Author
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Rai A, Vang D, Ritt M, and Sivaramakrishnan S
- Subjects
- Actin Cytoskeleton chemistry, Actin Cytoskeleton ultrastructure, Adaptor Proteins, Signal Transducing genetics, Apoptosis Regulatory Proteins genetics, Apoptosis Regulatory Proteins ultrastructure, Clathrin-Coated Vesicles chemistry, Clathrin-Coated Vesicles genetics, Cytoskeleton chemistry, Cytoskeleton genetics, Cytoskeleton ultrastructure, Endocytosis genetics, Endosomes genetics, Humans, Kinetics, Multiprotein Complexes genetics, Multiprotein Complexes ultrastructure, Myosin Heavy Chains genetics, Myosin Heavy Chains ultrastructure, Phosphatidylserines genetics, Protein Binding genetics, Protein Multimerization genetics, Single Molecule Imaging, Actin Cytoskeleton genetics, Adaptor Proteins, Signal Transducing chemistry, Apoptosis Regulatory Proteins chemistry, Multiprotein Complexes chemistry, Myosin Heavy Chains chemistry
- Abstract
Myosin VI ensembles on endocytic cargo facilitate directed transport through a dense cortical actin network. Myosin VI is recruited to clathrin-coated endosomes via the cargo adaptor Dab2. Canonically, it has been assumed that the interactions between a motor and its cargo adaptor are stable. However, it has been demonstrated that the force generated by multiple stably attached motors disrupts local cytoskeletal architecture, potentially compromising transport. In this study, we demonstrate that dynamic multimerization of myosin VI-Dab2 complexes facilitates cargo processivity without significant reorganization of cortical actin networks. Specifically, we find that Dab2 myosin interacting region (MIR) binds myosin VI with a moderate affinity (184 nM) and single-molecule kinetic measurements demonstrate a high rate of turnover (1 s
-1 ) of the Dab2 MIR-myosin VI interaction. Single-molecule motility shows that saturating Dab2-MIR concentration (2 μM) promotes myosin VI homodimerization and processivity with run lengths comparable with constitutive myosin VI dimers. Cargo-mimetic DNA origami scaffolds patterned with Dab2 MIR-myosin VI complexes are weakly processive, displaying sparse motility on single actin filaments and "stop-and-go" motion on a cellular actin network. On a minimal actin cortex assembled on lipid bilayers, unregulated processive movement by either constitutive myosin V or VI dimers results in actin remodeling and foci formation. In contrast, Dab2 MIR-myosin VI interactions preserve the integrity of a minimal cortical actin network. Taken together, our study demonstrates the importance of dynamic motor-cargo association in enabling cargo transportation without disrupting cytoskeletal organization., Competing Interests: Conflict of interests The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)- Published
- 2021
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