tailieunhanh - Báo cáo khoa học: Activation of transiently transfected reporter genes in 3T3 Swiss cells by the inducers of differentiation/apoptosis – dimethylsulfoxide, hexamethylene bisacetamide and trichostatin A

Despitedecadesof investigation, theprimary siteof actionof the prototypical inducers of differentiation, dimethylsulfox-ide andhexamethylene bisacetamide (HMBA), has not been studies designed to analyzecis-acting elements responsible for induction of stage-specific globin genes, we discovered the capacity of HMBA and dimethyl-sulfoxide toenhance the expressionof transiently transfected reporter genes derived from globin and nonglobin gene promoters, prominently innonerythroid3T3Swiss cells | Eur. J. Biochem. 271 2379-2390 2004 FEBS 2004 doi Activation of transiently transfected reporter genes in 3T3 Swiss cells by the inducers of differentiation apoptosis - dimethylsulfoxide hexamethylene bisacetamide and trichostatin A Kimiko Ishiguro and Alan C. Sartorelli Department of Pharmacology and Developmental Therapeutics Program Cancer Center Yale University School of Medicine New Haven CT USA Despite decades of investigation the primary site of action of the prototypical inducers of differentiation dimethylsulfoxide and hexamethylene bisacetamide HMBA has not been delineated. Durigg stddies desigeed to aanlyee cis-acting elements responsible for induction of stage-specific globin genes we discovered the capacity of HMBA and dimethylsulfoxide to enhance the expression of transiently transfected reporter genes derived from globin and nonglobin gene promoters prominently in nonerythroid 3T3 Swiss cells. The action of HMBA and dimethylsulfoxide in the transient transfection system resembled that of the inhibitor of histone deacetylases HDACs trichostatin A TSA in that the three agents enhanced reporter gene expression a regardless of the promoter employed b with similar kinetics and c with an increase in the steady-state level of reporter mRNA. Traniíendy 11 111 0. 1 DNA was assembied rapidly into a chromatiniyed structure in 3T3 cells suggesting that transcription of reporter genes was at least in part repressed by chromatin organization. Nudear iuin-on I y s indicated that dimethylsulfoxide and HMBA enhanced transcriptional initiation of the reporter and p21 WAF1 Cip1 genes. nn conlnitl. TSA prod need neg He effects on nuclear run-on transcription of these genes. HMBA ndd dimethylsulfoxide did not change the acetylation phosphorylation or methylation status of histones and did not activate stably transfected these differences the three agents modulated the expression of common sets of cellular genes and induced .

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