title: Investigations of human myosin VI targeting using optogenetically controlled cargo loading.
abstract: Myosins play countless critical roles in the cell, each requiring it to be activated at a specific location and time. To control myosin VI with this specificity, we created an optogenetic tool for activating myosin VI by fusing the light-sensitiveAvena sativaphototropin1 LOV2 domain to a peptide from Dab2 (LOVDab), a myosin VI cargo protein. Our approach harnesses the native targeting and activation mechanism of myosin VI, allowing direct inferences on myosin VI function. LOVDab robustly recruits human full-length myosin VI to various organelles in vivo and hinders peroxisome motion in a light-controllable manner. LOVDab also activates myosin VI in an in vitro gliding filament assay. Our data suggest that protein and lipid cargoes cooperate to activate myosin VI, allowing myosin VI to integrate Ca2+, lipid, and protein cargo signals in the cell to deploy in a site-specific manner.
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637.
authors.forename: Alexander R
authors.initials: AR
authors.lastname: French
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637; trsosnic@uchicago.edu rrock@uchicago.edu.|Institute for Biophysical Dynamics, The University of Chicago, Chicago, IL 60637.
authors.forename: Tobin R
authors.initials: TR
authors.lastname: Sosnick
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL 60637; trsosnic@uchicago.edu rrock@uchicago.edu.|Institute for Biophysical Dynamics, The University of Chicago, Chicago, IL 60637.
authors.forename: Ronald S
authors.initials: RS
authors.lastname: Rock
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D006367:HeLa Cells; D006801:Humans; D018995:Myosin Heavy Chains / Q000235:genetics* / Q000378:metabolism*; D062308:Optogenetics / Q000379:methods; D015290:Second Messenger Systems / Q000235:genetics
chemical_list: D000199:Actins; C089785:myosin VI; D018995:Myosin Heavy Chains
keywords: LOV2; motor proteins; myosin VI; optogenetics; protein regulation