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Single Cell Analysis of Human RAD18-Dependent DNA Post-Replication Repair by Alkaline Bromodeoxyuridine Comet Assay

Mórocz, Mónika and Gali, Himabindu and Raskó, István and Haracska, Lajos and Downes, C. Stephen (2013) Single Cell Analysis of Human RAD18-Dependent DNA Post-Replication Repair by Alkaline Bromodeoxyuridine Comet Assay. PLOS ONE, 8 (8). ISSN 1932-6203

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Abstract

Damage to DNA can block replication progression resulting in gaps in the newly synthesized DNA. Cells utilize a number of post-replication repair (PRR) mechanisms such as the RAD18 controlled translesion synthesis or template switching to overcome the discontinuities formed opposite the DNA lesions and to complete DNA replication. Gaining more insights into the role of PRR genes promotes better understanding of DNA damage tolerance and of how their malfunction can lead to increased genome instability and cancer. However, a simple and efficient method to characterise gene specific PRR deficiencies at a single cell level has not been developed. Here we describe the so named BrdU comet PRR assay to test the contribution of human RAD18 to PRR at a single cell level, by which we kinetically characterized the consequences of the deletion of human RAD18 on the replication of UV-damaged DNA. Moreover, we demonstrate the capability of our method to evaluate PRR at a single cell level in unsynchronized cell population. © 2013 Mórocz et al.

Item Type: Article
Uncontrolled Keywords: ultraviolet radiation; single cell analysis; POSTREPLICATION REPAIR; intermethod comparison; human cell; human; DNA Replication; DNA Repair; DNA Damage; controlled study; comet assay; cell labeling; ARTICLE; Rad18 protein; broxuridine
Subjects: Q Science / természettudomány > QH Natural history / természetrajz > QH301 Biology / biológia
R Medicine / orvostudomány > R1 Medicine (General) / orvostudomány általában
SWORD Depositor: MTMT SWORD
Depositing User: MTMT SWORD
Date Deposited: 15 Oct 2013 14:31
Last Modified: 15 Oct 2013 14:31
URI: http://real.mtak.hu/id/eprint/6896

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