RcLEA, a late embryogenesis abundant protein gene isolated from Rosa chinensis, confers tolerance to Escherichia coli and Arabidopsis thaliana and stabilizes enzyme activity under diverse stresses.
The late embryogenesis abundant (LEA) protein family is a large protein family that is closely associated with resistance to abiotic stresses in many organisms, such as plants, bacteria and animals. In this study, we isolated a LEA gene, RcLEA, which was cytoplasm-localized, from Rosa chinensis. RcLEA was found to be induced by high temperature through RT-PCR. Overexpression of RcLEA in Escherichia coli improved its growth performance compared with the control under high temperature, low temperature, NaCl and oxidative stress conditions. RcLEA was also overexpressed in Arabidopsis thaliana. The transgenic Arabidopsis showed better growth after high and low temperature treatment and exhibited less peroxide according to 3, 3-diaminobenzidine staining. However, RcLEA did not improve the tolerance to NaCl or osmotic stress in Arabidopsis. In vitro analysis showed that RcLEA was able to prevent the freeze-thaw-induced inactivation or heat-induced aggregation of various substrates, such as lactate dehydrogenase and citrate synthase. It also protected the proteome of E. coli from denaturation when the proteins were heat-shocked or subjected to acidic conditions. Furthermore, bimolecular fluorescence complementation assays suggested that RcLEA proteins function in a complex manner by making the form of homodimers.
State Key Laboratory of Genetic Engineering, Institute of Genetics, Fudan University, 220 Handan Road, Shanghai, 200433, China.
Xuan
X
Zhang
Songchong
S
Lu
Changhua
C
Jiang
Yaofeng
Y
Wang
Bo
B
Lv
Jiabin
J
Shen
Feng
F
Ming
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics*; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D018506:Gene Expression Regulation, Plant / Q000502:physiology*; D006358:Hot Temperature; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D050296:Microbial Viability; D018384:Oxidative Stress; D010802:Phylogeny; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism; D027826:Rosa / Q000235:genetics / Q000378:metabolism*; D012965:Sodium Chloride; D013312:Stress, Physiological / Q000235:genetics; D015854:Up-Regulation
D010940:Plant Proteins; D018749:RNA, Plant; D012965:Sodium Chloride; D007770:L-Lactate Dehydrogenase