کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10738958 | 1046849 | 2005 | 11 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Identification of an E-box motif as a transcriptional repressor element in the proximal promoter region of the GCLC gene in rat lung epithelial L2 cells
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کلمات کلیدی
NF1EMSAbHLHZipAP-4MZF1GCLMGCLCARNTNF-κBAP-1GCLHRPPBSFBSDTTELFC/EBP - C / EBPSp1 - SP1aryl hydrocarbon receptor nuclear translocator - اتمسفر هسته ای گیرنده آرویل کربنsodium dodecyl sulfate-polyacrylamide gel electrophoresis - الکتروفورز ژل دوده سولفات سدیم پلی آکریل آمیدSDS-PAGE - الکتروفورز ژل پلی آکریل آمیدmyeloid zinc finger 1 - انگشت میلوئید روی 1Chronic obstructive pulmonary disease - بیماری مزمن انسدادی ریهCOPD - بیماری مزمن انسدادی ریهelectrophoretic mobility-shift assay - تحرک انتقال الکتریکی - تحرک تغییرOxidative stress - تنش اکسیداتیوdithiothreitol - دیتیوتریتولbasic helix-loop-helix leucine zipper - زیپ لوکسی اسلیمی اسلحه پایهfetal bovine serum - سرم جنین گاوnuclear factor 1 - عامل هسته ای 1antioxidant-response element - عنصر پاسخ آنتی اکسیدانیUpstream stimulatory factor - فاکتور تحرک بالاnuclear factor κB - فاکتور هسته ای κBglutamate-cysteine ligase - لیگاز گلوتامات سیتئینEpithelial lining fluid - مایع تخلیه اپیتلیالARE - هستندpolymerase chain reaction - واکنش زنجیره ای پلیمرازPCR - واکنش زنجیرهٔ پلیمرازHorseradish peroxidase - پراکسیداز هوررادیشStimulating protein 1 - پروتئین تحریک کننده 1activator protein 1 - پروتئین فعال کننده 1Glutathione - گلوتاتیون
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
سالمندی
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Identification of an E-box motif as a transcriptional repressor element in the proximal promoter region of the GCLC gene in rat lung epithelial L2 cells Identification of an E-box motif as a transcriptional repressor element in the proximal promoter region of the GCLC gene in rat lung epithelial L2 cells](/preview/png/10738958.png)
چکیده انگلیسی
Glutathione (GSH) is a critical antioxidant for protecting the airway epithelium from oxidant injury and its levels are mainly controlled by glutamate-cysteine ligase (GCL), which is the rate-limiting enzyme in GSH synthesis. A full understanding of the gene regulation mechanism of this important enzyme may disclose the role it plays in respiratory diseases. GCL is made up of two differentially regulated subunits, a catalytic or heavy subunit (GCLC) and a modifier or light subunit (GCLM). Many studies in this field led to the findings of important positive regulatory regions of the GCLC promoter. For a detailed analysis of this gene regulation in the respiratory system, we cloned a 1.76-kb 5â²-flanking region of the rat GCLC gene, inserted into a luciferase reporter vector. Exonuclease III was used to cut the 5â²-flanking region of the rat GCLC gene unidirectionally into deletion mutants of different lengths. Sequential deletion analysis revealed that regions from â403 to â111 and from â705 to â613 are involved in positive regulation and the region from â745 to â705 is involved in negative regulation of the GCL gene in rat lung epithelial L2 cells. Specific proteins binding to these regions were confirmed by electrophoretic mobility-shift assays (EMSAs) and antibody supershift assays. An E-box motif was found in the negative regulatory region â745 to â705. Site-directed mutagenesis proved that the functional element in this negative regulatory region was a putative E-box element. EMSA and supershift assays showed that USF1 and USF2 can specifically bind to the E-box element. Overexpression of USFs in L2 cells led to a decreased activity of the GCLC promoter. Western blotting demonstrated that the expression of GCLC protein was decreased in the retroviral USFs-expressing cells than in nontransfected (no DNA added) cells, suggesting that USF binding to the E-box at â729/â724 serves to trans-repress GCLC gene expression. These findings indicate that the E-box is an important transcriptional suppressor element in the GCLC promoter in rat lung epithelial L2 cells. Inhibition of interaction between the E-box and the USF may provide an effective means of ameliorating oxidant injury of the lung.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Free Radical Biology and Medicine - Volume 39, Issue 8, 15 October 2005, Pages 1030-1040
Journal: Free Radical Biology and Medicine - Volume 39, Issue 8, 15 October 2005, Pages 1030-1040
نویسندگان
Lin-Ling Cheng, Bing Li, Jian-Dong Luo, Hong-Bin Tu, Qi-Cai Liu, Pixin Ran,