title: Cross-species functional complementation of cellulose synthase during the development of cellular slime molds.
abstract: Cellulose is a major and important component of the extracellular matrix during the development of Dictyostelium discoideum. Upon starvation, solitary amoebae of D. discoideum gather and form fruiting bodies in which cells differentiate into stalk cells and spores. The stalk tubes and walls of spores and stalk cells are made of cellulose. In the genus Acytostelium, however, all cells are destined to become spores and the stalks comprise only a cellulose tube, suggesting species-specific regulation of cellulose synthesis. In this study, we cloned a putative cellulose synthase gene (cesA) of Acytostelium subglobosum and performed comparative analyses with the D. discoideum cellulose synthase gene (dcsA). Although the deduced amino acid sequences were highly conserved between cesA and dcsA, the numbers of transmembrane spans preceding the catalytic domain were dissimilar; 2 and 3, respectively. Since ectopic expression of cesA in dcsA(-) null cells failed to restore the developmental defects of the mutant, we constructed a series of chimerical genes for complementation analyses and found that the catalytic domain of cesA was functional in D. discoideum cells if the preceding transmembrane region was swapped with dcsA. The non-functional products that contained the cesA-derived transmembrane region were localized to lysosomes. These results indicate that the transmembrane region of cellulose synthase is essential for proper accumulation of cellulose during the development of D. discoideum and that its differential localization in A. subglobosum may be related to the characteristic morphogenesis in this species.
authors.affiliation: Faculty of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8572, Japan.
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Kuwayama
authors.forename: Takeru
authors.initials: T
authors.lastname: Tohyama
authors.forename: Hideko
authors.initials: H
authors.lastname: Urushihara
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002482:Cellulose / Q000096:biosynthesis*; D003001:Cloning, Molecular; D017124:Conserved Sequence / Q000235:genetics; D016837:Dictyosteliida / Q000201:enzymology* / Q000254:growth & development*; D005816:Genetic Complementation Test; D005822:Genetic Vectors / Q000235:genetics; D005964:Glucosyltransferases / Q000235:genetics* / Q000378:metabolism; D015151:Immunoblotting; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009024:Morphogenesis / Q000235:genetics / Q000502:physiology*; D012098:Reproduction / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013045:Species Specificity
chemical_list: D002482:Cellulose; D005964:Glucosyltransferases; C478648:cellulose synthase
keywords: Acytostelium subglobosum; Dictyostelium discoideum; cellulose synthesis; development