title: Enhancing stress tolerance by overexpression of a methionine sulfoxide reductase A (MsrA) gene in Pleurotus ostreatus.
abstract: Proteins are subjected to modification by reactive oxygen species (ROS), and oxidation of specific amino acid residues can impair their biological functions. Methionine as a sulfur-containing amino acid is easily oxidized to methionine sulfoxide (MetSO). The modified methionine can be repaired by methionine sulfoxide reductase (Msr), an enzyme that reverses oxidation of methionine in proteins. In this study, a methionine sulfoxide reductase A (PoMsrA) gene from Pleurotus ostreatus was cloned and characterized. Furthermore, the function of PoMsrA gene was analyzed by overexpression in P. ostreatus via Agrobacterium-mediated transformation. Stable integration of the target gene into the genome of P. ostreatus was confirmed by PCR, fluorescence observation, and Southern blot hybridization. qRT-PCR analysis showed that PoMsrA was highly expressed in the stage of mature and young fruiting bodies as well as the osmotic stress condition of 0.3 M NaCl. Additionally, the transgenic strains with PoMsrA overexpression exhibited an enhanced tolerance to high temperature, high osmotic stress, and oxidative stress. This suggests that PoMsrA is an active player in the protection of the cellular proteins from oxidative stress damage.
authors.affiliation: College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China.
authors.forename: Chaomin
authors.initials: C
authors.lastname: Yin
authors.forename: Liesheng
authors.initials: L
authors.lastname: Zheng
authors.forename: Jihong
authors.initials: J
authors.lastname: Zhu
authors.forename: Liguo
authors.initials: L
authors.lastname: Chen
authors.forename: Aimin
authors.initials: A
authors.lastname: Ma
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D051150:Methionine Sulfoxide Reductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D010802:Phylogeny; D020076:Pleurotus / Q000235:genetics / Q000254:growth & development / Q000502:physiology*; D011487:Protein Conformation; D016415:Sequence Alignment; D013312:Stress, Physiological
chemical_list: D005656:Fungal Proteins; D051150:Methionine Sulfoxide Reductases; C027219:methionine sulfoxide reductase