Monalisa Mukherjea, M. Yusuf Ali, Carlos Kikuti, Daniel Safer, Zhaohui Yang, Helena Sirkia, Virginie Ropars, Anne Houdusse, David M. Warshaw, H. Lee Sweeney
2014 vol: 8 issue: 5 pp: 1522-1532 doi: 10.1016/j.celrep.2014.07.041
It is unclear whether the reverse-direction myosin (myosin VI) functions as a monomer or dimer in cells and how it generates large movements on actin. We deleted a stable, single-α-helix (SAH) domain that has been proposed to function as part of a lever arm to amplify movements without impact on in vitro movement or in vivo functions. A myosin VI construct that used this SAH domain as part of its lever arm was able to take large steps in vitro but did not rescue in vivo functions. It was necessary for myosin VI to internally dimerize, triggering unfolding of a three-helix bundle and calmodulin binding in order to step normally in vitro and rescue endocytosis and Golgi morphology in myosin VI-null fibroblasts. A model for myosin VI emerges in which cargo binding triggers dimerization and unfolds the three-helix bundle to create a lever arm essential for in vivo functions.
Topics: Binding mechanism, Peptides, Monolith – MicroScale Thermophoresis, MST, Proteins, Publications