title: Engineered myosin VI motors reveal minimal structural determinants of directionality and processivity.
abstract: Myosins have diverse mechanical properties reflecting a range of cellular roles. A major challenge is to understand the structural basis for generating novel functions from a common motor core. Myosin VI (M6) is specialized for processive motion toward the (-) end of actin filaments. We have used engineered M6 motors to test and refine the "redirected power stroke" model for (-) end directionality and to explore poorly understood structural requirements for processive stepping. Guided by crystal structures and molecular modeling, we fused artificial lever arms to the catalytic head of M6 at several positions, retaining varying amounts of native structure. We found that an 18-residue alpha-helical insert is sufficient to reverse the directionality of the motor, with no requirement for any calmodulin light chains. Further, we observed robust processive stepping of motors with artificial lever arms, demonstrating that processivity can arise without optimizing lever arm composition or mechanics.
authors.affiliation: Department of Bioengineering, Stanford University, Stanford, CA 94305, USA.
authors.forename: Jung-Chi
authors.initials: JC
authors.lastname: Liao
authors.forename: Mary Williard
authors.initials: MW
authors.lastname: Elting
authors.forename: Scott L
authors.initials: SL
authors.lastname: Delp
authors.forename: James A
authors.initials: JA
authors.lastname: Spudich
authors.forename: Zev
authors.initials: Z
authors.lastname: Bryant
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001696:Biomechanical Phenomena; D003198:Computer Simulation; D007700:Kinetics; D008958:Models, Molecular; D020409:Molecular Motor Proteins / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D008969:Molecular Sequence Data; D009068:Movement / Q000502:physiology*; D018995:Myosin Heavy Chains / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering; D055503:Protein Multimerization / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D017386:Sequence Homology, Amino Acid
chemical_list: D020409:Molecular Motor Proteins; C089785:myosin VI; D018995:Myosin Heavy Chains