Isolation and characterization of a metallothionein-1 protein in Chloris virgata Swartz that enhances stress tolerances to oxidative, salinity and carbonate stress in Saccharomyces cerevisiae.
Chloris virgata Swartz (C. virgata) is a gramineous wild plant that is found in alkaline soil areas in northeast China and is highly tolerant to carbonate stress. We constructed a cDNA library from C. virgata seedlings treated with NaHCO(3), and isolated a type 1 metallothionein (MT1) gene (ChlMT1: AB294238) from the library. The amino acid sequence of ChlMT1 contained 12 cysteine residues that constituted the Cys-X-Cys (X = amino acid except Cys) motifs in the N- and C-terminal regions. Northern hybridization showed that expression of ChlMT1 was induced by several abiotic stresses, from salts (NaCl and NaHCO(3)), a ROS inducer (paraquat), and metals (CuSO(4), ZnSO(4), and CoCl(2)). ChlMT1 expression in leaf was induced by 200 mM NaCl and 100 mM NaHCO(3). About 5 microM Paraquat, 500 microM Zn(2+), and 500 microM Co(2+) also induced expression of ChlMT1 in leaf after 6 h, and 100 microM Cu(2+) induced it after 24 h. Saccharomyces cerevisiae when transformed with the ChlMT1 gene had dramatically increased tolerances to salts (NaCl and NaHCO(3)) and ROS.
Nishiuchi
Shunsaku
S
Asian Natural Environmental Science Center, The University of Tokyo, 1-1-1, Midori-cho, Nishitokyo-shi, Tokyo, 188-0002, Japan.
Liu
Shenkui
S
Takano
Tetsuo
T
D000595:Amino Acid Sequence; D002244:Carbon / Q000737:chemistry; D008668:Metallothionein / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D008957:Models, Genetic; D008969:Molecular Sequence Data; D018384:Oxidative Stress; D010100:Oxygen / Q000378:metabolism; D018515:Plant Leaves / Q000378:metabolism; D010944:Plants / Q000378:metabolism*; D017382:Reactive Oxygen Species; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D012492:Salts / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D012965:Sodium Chloride / Q000737:chemistry
D017382:Reactive Oxygen Species; D012492:Salts; D012965:Sodium Chloride; D002244:Carbon; D008668:Metallothionein; D010100:Oxygen