title: Overexpression of Hsp90 from grass carp (Ctenopharyngodon idella) increases thermal protection against heat stress.
abstract: With homologous DNA probes, we had screened a grass carp heat shock protein 90 gene (CiHsp90). The full sequence of CiHsp90 cDNA was 2793 bp, which could code a 798 amino acids peptide. The phylogenetic analysis demonstrated that CiHsp90 shared the high homology with Zebrafish Grp94. Quantitative RT-PCR analysis showed that CiHsp90 was ubiquitously expressed at lower levels in all detected tissues and up-regulated after heat shock at 34 °C or cold stress at 4 °C. To understand the function of CiHsp90 involving in thermal protection, an expression vector containing coding region cDNA was expressed in E. coli BL21 (DE3) plysS. Upon transfer from 37 °C to 42 °C, these cells that accumulated CiHsp90 peptides displayed greater thermoresistance than the control cells. While incubated at 4°C for different periods, it could also improve the cell viability. After transient transfected recombinant plasmid pcDNA3.1/CiHsp90 into mouse myeloma cell line SP2/0, we found that CiHsp90 could contribute to protecting cells against both thermal and cold extremes. On the contrary, the mutant construct ΔN-CiHsp90 (256-798aa) could abolish the protection activity both in prokaryotic cells and eukaryotic cells. Additionally, both CiHsp90 and ΔN-CiHsp90 peptides could reduce the level of citrate synthase aggregation at the high temperature.
authors.affiliation: Nanchang Teachers College, Nanchang 330103, China.
authors.forename: Chu-Xin
authors.initials: CX
authors.lastname: Wu
authors.forename: Feng-Yun
authors.initials: FY
authors.lastname: Zhao
authors.forename: Yuan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Yu-Jiao
authors.initials: YJ
authors.lastname: Zhu
authors.forename: Mei-Sheng
authors.initials: MS
authors.lastname: Ma
authors.forename: Hui-Ling
authors.initials: HL
authors.lastname: Mao
authors.forename: Cheng-Yu
authors.initials: CY
authors.lastname: Hu
mesh_terms: D000818:Animals; D002347:Carps / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002950:Citrate (si)-Synthase / Q000235:genetics; D003080:Cold Temperature; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D020869:Gene Expression Profiling; D018841:HSP90 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D006358:Hot Temperature; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D010802:Phylogeny; D017386:Sequence Homology, Amino Acid; D013312:Stress, Physiological / Q000235:genetics
chemical_list: D018841:HSP90 Heat-Shock Proteins; D002950:Citrate (si)-Synthase