Transcriptional regulation involving the intronic heat shock element of the rat hsp27 gene
| Autor: | Lyndon F. Cooper, Zhanying Guo, Katsumi Uoshima |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
endocrine system
animal structures Transcription Genetic Response element Restriction Mapping Biophysics HSP27 Heat-Shock Proteins Biology Transfection Biochemistry Cell Line HSPA1B Structural Biology Transcription (biology) Genes Reporter Heat shock protein Genetics Animals HSF1 Promoter Regions Genetic Heat-Shock Proteins HSPA12A Temperature Nuclear Proteins Molecular biology Introns Neoplasm Proteins Rats Heat shock factor Open reading frame embryonic structures |
| Zdroj: | Biochimica et biophysica acta. 1490(3) |
| ISSN: | 0006-3002 |
| Popis: | While sequencing the first intron of the rat heat shock protein 27 gene (hsp27), we identified a consensus heat shock regulatory element (HSE). This intronic HSE (i-HSE) is conserved among mammalian hsp27 genes. The aim of this study was to investigate possible effects of this intronic HSE (i-HSE) on transcription from the rat hsp27 promoter. Gel mobility shift assays indicated that the i-HSE bound heat shock transcription factor 1 (HSF1) in a manner equivalent to that of HSE present in hsp27 promoter (p-HSE). The effect of i-HSE on transcription from the hsp27 promoter was evaluated using reporter constructs transiently transfected in the osteosarcoma cell line ROS17/2.8. When inserted 5' to a 145 bp fragment of the hsp27 promoter not containing p-HSE, a 215 bp fragment of hsp27 intron 1 containing i-HSE enhanced CAT activity and conferred heat shock-inducible activity to the construct. This intronic fragment containing i-HSE also enhanced CAT activity in either normal or heat-shocked culture conditions when inserted 3' to the CAT open reading frame. However, in chimeric reporter constructs with a 273 bp hsp27 promoter containing p-HSE directly 5' to CAT reporter, and with a 215 bp fragment containing i-HSE inserted 3' to the CAT open reading frame, transcription from hsp27 promoter was reduced under normal and heat-stressed culture conditions. Mutation of the i-HSE reversed this effect. Further study is required to define the mechanism by which the i-HSE-containing region of the hsp27 promoter may mediate negative regulation of hsp27 transcription. |
| Databáze: | OpenAIRE |
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