IDENTIFICATION OF A 2ND REGION UPSTREAM OF THE MOUSE HEME OXYGENASE-1 GENE THAT FUNCTIONS AS A BASAL LEVEL AND INDUCER-DEPENDENT TRANSCRIPTION ENHANCER

被引:191
作者
ALAM, J
CAMHI, S
CHOI, AMK
机构
[1] LOUISIANA STATE UNIV,MED CTR,DEPT BIOCHEM & MOLEC BIOL,NEW ORLEANS,LA 70121
[2] JOHNS HOPKINS UNIV,SCH MED,DEPT MED,BALTIMORE,MD 21205
关键词
D O I
10.1074/jbc.270.20.11977
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A 161-base pair fragment (AB1) similar to 10 kilobase pairs upstream of the transcription start site of the mouse heme oxygenase-1 gene functions as a basal level and inducer-dependent enhancer. AB1/chloramphenicol acetyltransferase fusion genes stably transfected into mouse hepatoma (Hepa) cells or L929 fibroblasts were activated 7-8- or 17-22-fold, respectively, after treatment of the cells with either CdCl2 or heme. The AB1 fragment is composed largely of three tandem repeats containing two conserved core elements, A and B. Part of core element A (TCCGGAGCTGTG) resembles the consensus-binding site for transcription factor AP-4, whereas core element B (GCTGAGTCANGG) includes the consensus-binding site (TGAGTCA) for the AP-1 family of transcription factors, Nuclear proteins from Hepa cells did not bind to any of the core A elements, but bound to all three copies of the core B element, AB1 derivatives with one or two mutant AP-1-binding elements exhibited reduced but measurable inducer-dependent enhancer activity, but mutation of all three AP-1-binding sites abolished activation by CdCl2, and heme and also by mercury chloride, zinc chloride, H2O2, sodium arsenate, and 12-O-tetradecanoylphorbol-13-acetate. etate. Pretreatment of stably transfected L929 cells with protein kinase C inhibitors, but not with tyrosine kinase inhibitors or N-acetylcysteine, abrogated 12-O-tetradecanoylphorbol-13-acetate-dependent activation of the AB1/chloramphenicol acetyltransferase fusion gene. Induction by H2O2 was unaffected by the kinase inhibitors, but completely abolished by N-acetylcysteine. Heme-dependent induction was not significantly affected by any of these chemicals.
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收藏
页码:11977 / 11984
页数:8
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