| A non-isotopic assay uses bromouridine and RNA synthesis to detect DNA damage responses. | |
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MedLine Citation:
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PMID: 20394837 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Individuals with inherited xeroderma pigmentosum (XP) disorder and Cockayne syndrome (CS) are deficient in nucleotide excision repair and experience hypersensitivity to sunlight. Although there are several diagnostic assays for these disorders, the recovery of RNA synthesis (RRS) assay that can discriminate between XP cells and CS cells is very laborious. Here, we report on a novel non-radioisotope RRS assay that uses bromouridine (a uridine analog) as an alternative to (3)H-uridine. This assay can easily detect RNA polymerase I transcription in nucleoli and RNA polymerase II transcription in nuclei. The non-RI RSS assay also can rapidly detect normal RRS activity in HeLa cells. Thus, this assay is useful as a novel and easy technique for CS diagnosis. Because RRS is thought to be related to transcription-coupled DNA repair, which is triggered by the blockage of transcriptional machinery by DNA lesions, this assay may be of use for analysis of DNA repair, transcription, and/or genetic toxicity. |
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Authors:
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Mayu Hasegawa; Shigenori Iwai; Isao Kuraoka |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-04-13 |
Journal Detail:
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Title: Mutation research Volume: 699 ISSN: 0027-5107 ISO Abbreviation: Mutat. Res. Publication Date: 2010 Jun |
Date Detail:
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Created Date: 2010-05-25 Completed Date: 2010-08-20 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0400763 Medline TA: Mutat Res Country: Netherlands |
Other Details:
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Languages: eng Pagination: 62-6 Citation Subset: IM |
Copyright Information:
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Copyright 2010 Elsevier B.V. All rights reserved. |
Affiliation:
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Graduate School of Engineering Science, Osaka University, Toyonaka, Osaka 560-8531, Japan. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Biological Assay* Cell Line Cockayne Syndrome / genetics DNA Damage / radiation effects* DNA Repair Hela Cells Humans Microscopy, Fluorescence RNA / biosynthesis* RNA Polymerase I / metabolism RNA Polymerase II / metabolism Ultraviolet Rays / adverse effects Uridine / analogs & derivatives*, metabolism Xeroderma Pigmentosum / genetics |
| Chemical | |
Reg. No./Substance:
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58-96-8/Uridine; 63231-63-0/RNA; 957-75-5/5-bromouridine; EC 2.7.7.-/RNA Polymerase I; EC 2.7.7.-/RNA Polymerase II |
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