کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5907681 | 1160132 | 2008 | 10 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
NRF-1, and AP-1 regulate the promoter of the human calpain small subunit 1 (CAPNS1) gene
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کلمات کلیدی
RT-PCRNRF-1uPARCAPNS1Activator protein 1 (AP-1)Calpain small subunit 1Small interfering RNA - RNA تداخل کوچکsiRNA - siRNAchromatin immunoprecipitation - ایمن سازی کروماتینPromoter analysis - تجزیه و تحلیل سازندهTranscriptional regulation - تنظیم ترانزیتیreverse transcription PCR - رونویسی معکوس PCREMSA یا electrophoretic mobility shift assay - سنجش تغییر تحرک الکتروفورتیکNuclear respiratory factor 1 - عامل تنفسی هسته ای 1CHiP - چیپUrokinase plasminogen activator receptor - گیرنده فعال کننده پلاسمینوژن اورو کیناز
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
ژنتیک
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: NRF-1, and AP-1 regulate the promoter of the human calpain small subunit 1 (CAPNS1) gene NRF-1, and AP-1 regulate the promoter of the human calpain small subunit 1 (CAPNS1) gene](/preview/png/5907681.png)
چکیده انگلیسی
Ubiquitously expressed µ- and m-calpain are cysteine proteases with broad functions in cell spreading, migration, proliferation, apoptosis, and in tumor invasion. They are heterodimers, with a distinct large 80-kDa catalytic, and a common small 28-kDa regulatory subunit (Capn4/CAPNS1). CAPNS1 is required to maintain stability and activity of both calpains. Despite its biological importance, the transcriptional regulation of this gene has not been studied, and the CAPNS1 promoter has not yet been characterized. In this study, we identified the main transcriptional start site, and cloned and characterized the ~ 2.0 kb upstream region of the CAPNS1 gene. Deletion analysis identified the core promoter located within region â 187/+174. Site-directed mutagenesis, EMSA- and supershift analysis identified Sp1-, NRF-1-, and AP-1-binding elements within the CAPNS1 core promoter. Binding of NRF-1, Sp1 and AP-1 to the natural core promoter was confirmed by chromatin immunoprecipitation (ChIP). Site-directed mutagenesis at the NRF-1 site in HeLa and MCF7 cells substantially reduced core promoter activity by 70%, whereas mutation of the AP-1-binding and Sp1-binding site reduced promoter activity by 50% and 30%, respectively. Double mutation of the NRF-1 and the AP-1 site reduced promoter activity by 90%. In Drosophila SL2 cells, ectopic expression of NRF-1 led to a significant induction of CAPNS1 promoter activity. Furthermore, an siRNA against NRF-1 substantially reduced promoter activity in HeLa cells, which was paralleled by a significant downregulation of CAPNS1 mRNA. These results reveal that especially NRF-1, along with AP-1 and, to a minor extent, an Sp1 site, is essential for human CAPNS1 promoter activity and gene expression.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Gene - Volume 410, Issue 1, 29 February 2008, Pages 197-206
Journal: Gene - Volume 410, Issue 1, 29 February 2008, Pages 197-206
نویسندگان
Irfan A. Asangani, Suhail A.K. Rasheed, Jörg H. Leupold, Stefan Post, Heike Allgayer,