title: A heat shock protein 90 β isoform involved in immune response to bacteria challenge and heat shock from Miichthys miiuy.
abstract: Heat shock protein 90 (HSP90) is highly conserved molecular chaperone that plays a critical role in cellular stress response. In this study, we reported the identification and functional analysis of a heat shock protein 90 gene from miiuy croaker (designated Mimi-HSP90). Mimi-HSP90 contained five conserved HSP90 protein family signatures and shared 89.6%-99.5% similarity with other known HSP90 β isoform. Homology analysis and structure comparison further indicated that Mimi-HSP90 should be β isoform member of the HSP90 family. The molecular evolutionary analysis showed that HSP90 was under an overall strong purifying select pressure among fish species. Mimi-HSP90 gene was constitutively expressed in ten examined tissues, and the expression level of liver was higher than in other tissues. The expression level of Mimi-HSP90 gene under bacterial infection and heat shock were analyzed by real-time quantitative RT-PCR, resulted in significant changes in liver, spleen, and kidney tissues. The purified recombinant pET-HSP90 protein was used to produce the polyclonal antibody in mice. The specificity of the antibody was determined by Western blot analysis. All results suggested that Mimi-HSP90 was involved in thermal stress and immune response in miiuy croaker.
authors.affiliation: Laboratory for Marine Living Resources and Molecular Engineering, College of Marine Science, Zhejiang Ocean University, 105 Wenhua Road, Zhoushan 316000, PR China.
authors.forename: Tao
authors.initials: T
authors.lastname: Wei
authors.forename: Yunhang
authors.initials: Y
authors.lastname: Gao
authors.forename: Rixin
authors.initials: R
authors.lastname: Wang
authors.forename: Tianjun
authors.initials: T
authors.lastname: Xu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D018841:HSP90 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D018869:Heat-Shock Response; D008969:Molecular Sequence Data; D010473:Perciformes / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment / Q000662:veterinary; D014733:Vibrio / Q000502:physiology*
chemical_list: D018076:DNA, Complementary; D018841:HSP90 Heat-Shock Proteins; D012333:RNA, Messenger
keywords: Evolutionary analysis; Gene expression; Heat shock protein 90 (HSP90); Miichthys miiuy (Miiuy croaker); Western blot