title: Evolutionarily conserved sites in yeast tropomyosin function in cell polarity, transport and contractile ring formation.
abstract: Tropomyosin is a coiled-coil protein that binds and regulates actin filaments. The tropomyosin gene in Schizosaccharomyces pombe, cdc8, is required for formation of actin cables, contractile rings, and polar localization of actin patches. The roles of conserved residues were investigated in gene replacement mutants. The work validates an evolution-based approach to identify tropomyosin functions in living cells and sites of potential interactions with other proteins. A cdc8 mutant with near-normal actin affinity affects patch polarization and vacuole fusion, possibly by affecting Myo52p, a class V myosin, function. The presence of labile residual cell attachments suggests a delay in completion of cell division and redistribution of cell patches following cytokinesis. Another mutant with a mild phenotype is synthetic negative with GFP-fimbrin, inferring involvement of the mutated tropomyosin sites in interaction between the two proteins. Proteins that assemble in the contractile ring region before actin do so in a mutant cdc8 strain that cannot assemble condensed actin rings, yet some cells can divide. Of general significance, LifeAct-GFP negatively affects the actin cytoskeleton, indicating caution in its use as a biomarker for actin filaments.
authors.affiliation: Department of Pathology and Laboratory Medicine, Robert Wood Johnson Medical School, Rutgers University, Piscataway, NJ 08854, USA.
authors.forename: Susanne
authors.initials: S
authors.lastname: Cranz-Mileva
authors.affiliation: Department of Pathology and Laboratory Medicine, Robert Wood Johnson Medical School, Rutgers University, Piscataway, NJ 08854, USA.
authors.forename: Brittany
authors.initials: B
authors.lastname: MacTaggart
authors.affiliation: Department of Pathology and Laboratory Medicine, Robert Wood Johnson Medical School, Rutgers University, Piscataway, NJ 08854, USA.
authors.forename: Jacquelyn
authors.initials: J
authors.lastname: Russell
authors.affiliation: Department of Pathology and Laboratory Medicine, Robert Wood Johnson Medical School, Rutgers University, Piscataway, NJ 08854, USA hitchcoc@rwjms.rutgers.edu.
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Hitchcock-DeGregori
keywords: Actin binding protein; Actin cytoskeleton; Fimbrin; Fission yeast; LifeAct; Myosin; Schizosaccharomyces pombe; cdc8