Short reports
ScienceAsia (): 253-257 |doi:
10.2306/scienceasia1513-1874...253
Temperature dependent γH2AX binding to DNA
Narisorn Kongruttanachoka, Chutipa Phuangphairojb, Wanpen Ponyeamb, Apiwat Mutirangurab*
ABSTRACT: γ-H2AX, the serine 139 phosphorylated form of histone H2AX, is one of the earliest DNA double-strand breaks repair responses. Therefore, the quantity of γ-H2AX-bound DNA usually reflects the extent of DNA damage and consequently, mutagenic potential. The purpose of this study was to assess the cellular evidence regarding whether and how temperature is related to mutation. This study applies our novel assay technique to measure the quantity of γ-H2AX-bound DNA by PCR. Chromatin immunoprecipitation technique was applied to select for the γ-H2AX-bound DNA complex. Then, the amount of precipitated DNA was measured by long interspersed element-1 (LINE-1) repetitive sequences real-time PCR. We observed a positive correlation between the amount of precipitated LINE-1 sequences and the exposure to temperature. The quantity of γ-H2AX-bound DNA increased significantly when HeLa was incubated in 47 °C for 30 min. In contrast, incubation at 4 °C for 30 min reduced the γ-H2AX-bound DNA quantity. Precise quantification of γ-H2AX-bound DNA will be a tool to prove whether there is a linkage between temperature and mutagenic effect.
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Inter-Department Program of Biomedical Sciences, Faculty of Graduate School, Chulalongkorn University, Bangkok 10330, Thailand. |
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Center of Excellence in Molecular Genetics of Cancer and Human Diseases, Department of Anatomy, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. |
* Corresponding author, E-mail: mapiwat@chula.ac.th
Received 4 Oct 2007, Accepted 30 Nov 2007
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