دورية أكاديمية
Protein-DNA footprinting of the human epsilon-globin promoter in human intact cells using nitrogen mustard analogues and other DNA-damaging agents.
| العنوان: | Protein-DNA footprinting of the human epsilon-globin promoter in human intact cells using nitrogen mustard analogues and other DNA-damaging agents. |
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| المؤلفون: | Temple MD; School of Biochemistry and Molecular Genetics, University of New South Wales, Sydney NSW 2052, Australia., Cairns MJ, Kim A, Murray V |
| المصدر: | Biochimica et biophysica acta [Biochim Biophys Acta] 1999 Jun 09; Vol. 1445 (3), pp. 245-56. |
| نوع المنشور: | Comparative Study; Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Pub. Co Country of Publication: Netherlands NLM ID: 0217513 Publication Model: Print Cited Medium: Print ISSN: 0006-3002 (Print) Linking ISSN: 00063002 NLM ISO Abbreviation: Biochim Biophys Acta Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Amsterdam : Elsevier Pub. Co. |
| مواضيع طبية MeSH: | DNA Damage* , Promoter Regions, Genetic*, DNA Footprinting/*methods , Globins/*genetics, Bleomycin/pharmacology ; DNA/chemistry ; DNA/drug effects ; Globins/chemistry ; Globins/metabolism ; HeLa Cells ; Humans ; K562 Cells ; Mechlorethamine/analogs & derivatives ; Mechlorethamine/pharmacology ; Polymerase Chain Reaction/methods ; Sulfuric Acid Esters/pharmacology |
| مستخلص: | Nitrogen mustard analogues, bleomycin and dimethyl sulphate (DMS) have been used as probes of protein-DNA interactions in intact human cells. The sites of damage have been determined at base pair resolution in the single copy epsilon-globin gene promoter in erythroid K562 cells, non-erythroid HeLa cells and purified DNA. Exponential amplification of gene-specific damage fragments was achieved using the ligation-mediated polymerase chain reaction (LMPCR) technique and analysed on DNA sequencing gels. A comparison of the relative damage band intensities between purified DNA and intact cells revealed several significant differences - both protection (footprint) and enhancement. These differences occurred at putative transcription factor binding sites and hence are thought to be due to protein-DNA interactions. A major feature of the band intensity ratio plots was the footprint observed at the CCAAT box binding motif as revealed by nitrogen mustard analogues. Enhanced band intensity (hypersensitivity) was displayed at the 5'- and 3'-ends of the CCAAT box in K562 cells - this feature was absent in HeLa cells and in vitro reconstitutions. A footprint was found at the GATA-1 motif in K562 cells that was also absent in non-expressing HeLa cells. Footprints were also evident at the TATA box, CACC box and the epsilonF1 DNA binding motif in K562 cells. |
| المشرفين على المادة: | 0 (Sulfuric Acid Esters) 11056-06-7 (Bleomycin) 50D9XSG0VR (Mechlorethamine) 9004-22-2 (Globins) 9007-49-2 (DNA) JW5CW40Z50 (dimethyl sulfate) |
| تواريخ الأحداث: | Date Created: 19990615 Date Completed: 19990719 Latest Revision: 20190610 |
| رمز التحديث: | 20240627 |
| DOI: | 10.1016/s0167-4781(99)00057-3 |
| PMID: | 10366709 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 0006-3002 |
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| DOI: | 10.1016/s0167-4781(99)00057-3 |