Dynamic multimerization of Dab2-Myosin VI complexes regulates cargo processivity while minimizing cortical actin reorganization

J Biol Chem. 2021 Jan-Jun:296:100232. doi: 10.1074/jbc.RA120.012703. Epub 2021 Jan 7.

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.

Keywords: Dab2; actin remodeling; cargo–motor interaction; motor processivity; myosin VI.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Actin Cytoskeleton / chemistry
  • Actin Cytoskeleton / genetics*
  • Actin Cytoskeleton / ultrastructure
  • Adaptor Proteins, Signal Transducing / chemistry*
  • Adaptor Proteins, Signal Transducing / genetics
  • Apoptosis Regulatory Proteins / chemistry*
  • 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 / chemistry*
  • Multiprotein Complexes / genetics
  • Multiprotein Complexes / ultrastructure
  • Myosin Heavy Chains / chemistry*
  • Myosin Heavy Chains / genetics
  • Myosin Heavy Chains / ultrastructure
  • Phosphatidylserines / genetics
  • Protein Binding / genetics
  • Protein Multimerization / genetics
  • Single Molecule Imaging

Substances

  • Adaptor Proteins, Signal Transducing
  • Apoptosis Regulatory Proteins
  • DAB2 protein, human
  • Multiprotein Complexes
  • Phosphatidylserines
  • myosin VI
  • Myosin Heavy Chains