TAILIEUCHUNG - Báo cáo khóa học: Cloning and characterization of two distinct isoforms of rainbow trout heat shock factor 1

To elucidate the molecular mechanism underlying the heat shock response in cold-water fish species, genes enco-ding heat shock transcription factors (HSFs) were cloned fromRTG-2cellsof the rainbowtroutOncorhynchusmykiss. Consequently, two distinctHSF1genes, namedHSF1aand HSF1b, were identified. The predicted amino acid sequence ofHSF1ashows identity to that proteins contained the general structuralmotifs ofHSF1, . a DNA-binding domain, hydrophobic heptad repeats and nuclear localization signals. . | Eur. J. Biochem. 271 703-712 2004 FEBS 2004 doi Cloning and characterization of two distinct isoforms of rainbow trout heat shock factor 1 Evidence for heterotrimer formation Nobuhiko Ojima and Michiaki Yamashita Cell Biology Section Physiology and Molecular Biology Division National Research Institute of Fisheries Science Fisheries Research Agency Yokohama Japan To elucidate the molecular mechanism underlying the heat shock response in cold-water fish species genes encoding heat shock transcription factors HSFs were cloned from of the rainbow trout Oncorhynchus mykiss. Consequently two distinct HSF1 genes named HSF1a and HSF1b were identified. The predicted amino acid sequence of HSF1a shows identity to that of HSF1b. The two proteins contained the general structural motifs of HSF1 . a DNA-binding domain hydrophobic heptad repeats and nuclear localization signals. Southern blot analysis showed that each HSF1 is encoded by a distinct gene. The two HSF1 mRNAs were coexpressed in unstressed rainbow trout RTG-2 cells nnd in various tissues. In nn electrophoretic mobility shift assay each in vitro translated HSF1 bound to the heat shock element. Chemical cross-linking and immunoprecipitation analysis showed that HSF1a and HSF1b form heterotrimers as well as homotrimers. Taken together these results demonstrate that in rainbow trout cells there are two distinct HSF1 isoforms that can form heterotrimers suggesting that a unique molecular mechanism underlies the stress response in tetraploid and or cold-water fish species. Keywords heat shock factor HSF1 isofoom rainbow trout trimerization. Heat shock proteins HSPs are highly conserved among a wide range of animals and are induced by environmental stressors such as elevated temperature heavy metals and oxidants. Many kinds of HSP have been reported to act as molecular chaperones that aid in the folding assembly degradation and translocation of intracellular proteins 1 .

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