Overexpression of bacterioferritin comigratory protein (Bcp) enhances viability and reduced glutathione level in the fission yeast under stress.
The structural gene encoding bacterioferritin comigratory protein (Bcp) was amplified using PCR from the genomic DNA of Schizosaccharomyces pombe, and transferred into the shuttle vector pRS316 to generate the recombinant plasmid pBCPlO. The bcp(+) mRNA level in the pBCPlO-containing yeast cells was significantly higher than that in the control yeast cells, indicating that the cloned gene is functioning. The S. pombe cells harboring the plasmid pBCPIO exhibited higher survival on the solid minimal media with hydrogen peroxide, tert-BOOH or cadmium than the control yeast cells. They also exhibited enhanced cellular viability in the liquid media containing the stressful agents. The increased viabilities of the fission yeast cells harboring the plasmid pBCP10 were also obtained with 0.4% glucose or 0.4% sucrose as a sole carbon source, and nitrogen starvation, compared with those of the control yeast cells. The total glutathione (GSH) content and total GSH/GSSG ratio were significantly higher in the yeast cells harboring the plasmid pBCP10 than in the control yeast cells. In brief, the S. pombe Bcp plays a protective role in the defensive response to oxidative stress possibly via up-regulation of total and reduced glutathione levels.
Kang
Ga-Young
GY
Division of Life Sciences and Research Institute of Life Sciences, Kangwon National University, Chuncheon 200-701, Republic of Korea.
Park
Eun-Hee
EH
Kim
Kyunghoon
K
Lim
Chang-Jin
CJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D016384:Consensus Sequence / Q000235:genetics; D004271:DNA, Fungal / Q000302:isolation & purification; D004274:DNA, Recombinant / Q000096:biosynthesis; D005978:Glutathione / Q000096:biosynthesis*; D050296:Microbial Viability / Q000235:genetics*; D008969:Molecular Sequence Data; D018384:Oxidative Stress / Q000235:genetics*; D054464:Peroxiredoxins / Q000096:biosynthesis* / Q000235:genetics*; D017382:Reactive Oxygen Species / Q000378:metabolism; D012568:Schizosaccharomyces / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D017422:Sequence Analysis, DNA; D015854:Up-Regulation
D004271:DNA, Fungal; D004274:DNA, Recombinant; D017382:Reactive Oxygen Species; D029702:Schizosaccharomyces pombe Proteins; D054464:Peroxiredoxins; C540496:SPBC1773.02c protein, S pombe; D005978:Glutathione