title: The functional expression and motile properties of recombinant outer arm dynein from Tetrahymena.
abstract: Cilia and flagella are motile organelles that play various roles in eukaryotic cells. Ciliary movement is driven by axonemal dyneins (outer arm and inner arm dyneins) that bind to peripheral microtubule doublets. Elucidating the molecular mechanism of ciliary movement requires the genetic engineering of axonemal dyneins; however, no expression system for axonemal dyneins has been previously established. This study is the first to purify recombinant axonemal dynein with motile activity. In the ciliated protozoan Tetrahymena, recombinant outer arm dynein purified from ciliary extract was able to slide microtubules in a gliding assay. Furthermore, the recombinant dynein moved processively along microtubules in a single-molecule motility assay. This expression system will be useful for investigating the unique properties of diverse axonemal dyneins and will enable future molecular studies on ciliary movement.
authors.affiliation: Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Tokyo, Japan. Electronic address: cedam@mail.ecc.u-tokyo.ac.jp.
authors.forename: Masaki
authors.initials: M
authors.lastname: Edamatsu
mesh_terms: D056444:Axonemal Dyneins / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D002923:Cilia / Q000502:physiology; D005407:Flagella / Q000502:physiology; D017125:Genes, Protozoan; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D008870:Microtubules / Q000502:physiology; D009068:Movement / Q000502:physiology; D015800:Protozoan Proteins / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013768:Tetrahymena / Q000737:chemistry / Q000235:genetics / Q000502:physiology*
chemical_list: D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D056444:Axonemal Dyneins
keywords: Cell motility; Ciliary movement; Cytoskeletal protein; Expression system; Recombinant axonemal dynein; Tetrahymena