共查询到20条相似文献,搜索用时 31 毫秒
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Wen SC Roder K Hu KY Rombel I Gavva NR Daftari P Kuo YY Wang C Shen CK 《Molecular and cellular biology》2000,20(6):1993-2003
The HS-40 enhancer is the major cis-acting regulatory element responsible for the developmental stage- and erythroid lineage-specific expression of the human alpha-like globin genes, the embryonic zeta and the adult alpha2/alpha/1. A model has been proposed in which competitive factor binding at one of the HS-40 motifs, 3'-NA, modulates the capability of HS-40 to activate the embryonic zeta-globin promoter. Furthermore, this modulation was thought to be mediated through configurational changes of the HS-40 enhanceosome during development. In this study, we have further investigated the molecular basis of this model. First, human erythroid K562 cells stably integrated with various HS-40 mutants cis linked to a human alpha-globin promoter-growth hormone hybrid gene were analyzed by genomic footprinting and expression analysis. By the assay, we demonstrate that factors bound at different motifs of HS-40 indeed act in concert to build a fully functional enhanceosome. Thus, modification of factor binding at a single motif could drastically change the configuration and function of the HS-40 enhanceosome. Second, a specific 1-bp, GC-->TA mutation in the 3'-NA motif of HS-40, 3'-NA(II), has been shown previously to cause significant derepression of the embryonic zeta-globin promoter activity in erythroid cells. This derepression was hypothesized to be regulated through competitive binding of different nuclear factors, in particular AP1 and NF-E2, to the 3'-NA motif. By gel mobility shift and transient cotransfection assays, we now show that 3'-NA(II) mutation completely abolishes the binding of small MafK homodimer. Surprisingly, NF-E2 as well as AP1 can still bind to the 3'-NA(II) sequence. The association constants of both NF-E2 and AP1 are similar to their interactions with the wild-type 3'-NA motif. However, the 3'-NA(II) mutation causes an approximately twofold reduction of the binding affinity of NF-E2 factor to the 3'-NA motif. This reduction of affinity could be accounted for by a twofold-higher rate of dissociation of the NF-E2-3'-NA(II) complex. Finally, we show by chromatin immunoprecipitation experiments that only binding of NF-E2, not AP1, could be detected in vivo in K562 cells around the HS-40 region. These data exclude a role for AP1 in the developmental regulation of the human alpha-globin locus via the 3'-NA motif of HS-40 in embryonic/fetal erythroid cells. Furthermore, extrapolation of the in vitro binding studies suggests that factors other than NF-E2, such as the small Maf homodimers, are likely involved in the regulation of the HS-40 function in vivo. 相似文献
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Bach1 functions as a hypoxia-inducible repressor for the heme oxygenase-1 gene in human cells 总被引:4,自引:0,他引:4
Kitamuro T Takahashi K Ogawa K Udono-Fujimori R Takeda K Furuyama K Nakayama M Sun J Fujita H Hida W Hattori T Shirato K Igarashi K Shibahara S 《The Journal of biological chemistry》2003,278(11):9125-9133
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Sun J Hoshino H Takaku K Nakajima O Muto A Suzuki H Tashiro S Takahashi S Shibahara S Alam J Taketo MM Yamamoto M Igarashi K 《The EMBO journal》2002,21(19):5216-5224
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Zenke-Kawasaki Y Dohi Y Katoh Y Ikura T Ikura M Asahara T Tokunaga F Iwai K Igarashi K 《Molecular and cellular biology》2007,27(19):6962-6971
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Nagai T Kikuchi S Ohmine K Miyoshi T Nakamura M Kondo T Furuyama K Komatsu N Ozawa K 《Journal of cellular biochemistry》2008,104(2):680-691
Heme plays an important biomodulating role in various cell functions. In this study, we examined the effects of hemin on cellular sensitivity to imatinib and other anti-leukemia reagents. Hemin treatment of human BCR/ABL-positive KCL22 leukemia cells increased IC(50) values of imatinib, that is, the drug resistance, in a dose-dependent manner without any change in the BCR/ABL kinase activity. Imatinib-induced apoptosis was also suppressed by hemin treatment in KCL22 cells. Hemin treatment increased the activity of gamma-glutamylcysteine synthetase (gamma-GCS) light subunit gene promoter, which contains a Maf recognition element (MARE). Protein levels of gamma-GCS and heme oxygenase-1 (HO-1), two MARE-containing genes, were also increased after hemin treatment. Knockdown of Nrf2 expression by RNA interference largely abolished the effect of hemin on imatinib-treated cells, suggesting that Nrf2 recognition of MARE is essential for the hemin-mediated protective effect. Similar to hemin, treatment of cells with delta-aminolevulinic acid (delta-ALA), the obligatory heme precursor, also increased IC(50) values of imatinib. In contrast, inhibition of cellular heme synthesis by succinylacetone increased the sensitivity of cells to imatinib in two imatinib-resistant cell lines, KCL22/SR and KU812/SR. Hemin treatment also decreased the sensitivity of cells to four anthracyclins, daunorubicin, idarubicin, doxorubicin, and mitoxantrone, in BCR/ABL-negative leukemia U937 and THP-1 cells, as well as in KCL22 cells. These findings thus indicate that cellular heme level plays an important role in determining the sensitivity of cells to imatinib and certain other anti-leukemia drugs and that the effect of heme may be mediated via its ability to upregulate Nrf2 activity. 相似文献
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Hoshino H Kobayashi A Yoshida M Kudo N Oyake T Motohashi H Hayashi N Yamamoto M Igarashi K 《The Journal of biological chemistry》2000,275(20):15370-15376
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Shuqing Zhang Minrui Xu Jian Huang Lili Tang Yanqing Zhang Jingyao Wu Shuo Lin Han Wang 《Disease models & mechanisms》2014,7(7):837-845
Using a zebrafish model of hepatoerythropoietic porphyria (HEP), we identify a previously unknown mechanism underlying heme-mediated regulation of exocrine zymogens. Zebrafish bach1b, nrf2a and mafK are all expressed in the zebrafish exocrine pancreas. Overexpression of bach1b or knockdown of nrf2a result in the downregulation of the expression of the exocrine zymogens, whereas overexpression of nrf2a or knockdown of bach1b cause their upregulation. In vitro luciferase assays demonstrate that heme activates the zymogens in a dosage-dependent manner and that the zymogen promoter activities require the integral Maf recognition element (MARE) motif. The Bach1b-MafK heterodimer represses the zymogen promoters, whereas the Nrf2a-MafK heterodimer activates them. Furthermore, chromatin immunoprecipitation (ChIP) assays show that MafK binds to the MARE sites in the 5′ regulatory regions of the zymogens. Taken together, these data indicate that heme stimulates the exchange of Bach1b for Nrf2a at MafK-occupied MARE sites and that, particularly in heme-deficient porphyria, the repressive Bach1b-MafK heterodimer dominates, which can be exchanged for the activating Nrf2a-MafK heterodimer upon treatment with hemin. These results provide novel insights into the regulation of exocrine function, as well as the pathogenesis of porphyria, and should be useful for designing new therapies for both types of disease.KEY WORDS: Bach1b, Nrf2a, MafK, Heme, Porphyria, Zymogens, Zebrafish 相似文献
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Redox regulation of the transcriptional repressor Bach1 总被引:4,自引:0,他引:4
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Identification of Bach2 as a B-cell-specific partner for small maf proteins that negatively regulate the immunoglobulin heavy chain gene 3'' enhancer. 下载免费PDF全文
A Muto H Hoshino L Madisen N Yanai M Obinata H Karasuyama N Hayashi H Nakauchi M Yamamoto M Groudine K Igarashi 《The EMBO journal》1998,17(19):5734-5743
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Cadmium induces nuclear export of Bach1, a transcriptional repressor of heme oxygenase-1 gene 总被引:5,自引:0,他引:5
Suzuki H Tashiro S Sun J Doi H Satomi S Igarashi K 《The Journal of biological chemistry》2003,278(49):49246-49253