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Cloning and characterization of Ku70 and Ku80 homologues involved in DNA repair process in wheat (Triticum aestivum L.)

Published online by Cambridge University Press:  16 July 2014

Jiayu Gu ,
Qing Wang ,
Meng Cui ,
Bing Han ,
Huijun Guo ,
Linshu Zhao ,
Yongdun Xie ,
Xiyun Song  and
Luxiang Liu
Jiayu Gu
Affiliation:
Institute of Crop Science, Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement/National Center of Space Mutagenesis for Crop Improvement, Beijing100081, People's Republic of China
Qing Wang
Affiliation:
Institute of Crop Science, Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement/National Center of Space Mutagenesis for Crop Improvement, Beijing100081, People's Republic of China College of Life Science, Qingdao Agricultural University, Qingdao266109, People's Republic of China
Meng Cui
Affiliation:
Institute of Crop Science, Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement/National Center of Space Mutagenesis for Crop Improvement, Beijing100081, People's Republic of China College of Life Science, Qingdao Agricultural University, Qingdao266109, People's Republic of China
Bing Han
Affiliation:
Institute of Crop Science, Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement/National Center of Space Mutagenesis for Crop Improvement, Beijing100081, People's Republic of China College of Life Science, Qingdao Agricultural University, Qingdao266109, People's Republic of China
Huijun Guo
Affiliation:
Institute of Crop Science, Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement/National Center of Space Mutagenesis for Crop Improvement, Beijing100081, People's Republic of China
Linshu Zhao
Affiliation:
Institute of Crop Science, Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement/National Center of Space Mutagenesis for Crop Improvement, Beijing100081, People's Republic of China
Yongdun Xie
Affiliation:
Institute of Crop Science, Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement/National Center of Space Mutagenesis for Crop Improvement, Beijing100081, People's Republic of China
Xiyun Song
Affiliation:
College of Life Science, Qingdao Agricultural University, Qingdao266109, People's Republic of China
Luxiang Liu*
Affiliation:
Institute of Crop Science, Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement/National Center of Space Mutagenesis for Crop Improvement, Beijing100081, People's Republic of China
*
* Corresponding author. E-mail: [email protected]
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Abstract

Error-prone repair of radiation-induced DNA double-strand breaks (DSBs) results in DNA mutation that is essential for mutation breeding. Non-homologous end joining might be the principal DSB repair mechanism in eukaryotes, which is mediated and activated by Ku protein, a heterodimer of 70 and 80 kDa subunits. In this study, on the basis of complementary DNA (cDNA), the genomic sequences of TaKu70 and TaKu80 genes in all the three genomes of wheat were characterized. Only single-nucleotide substitutions and no insertions or deletions were detected in the exons of TaKu70 and TaKu80 genes. The size of the introns exhibited a slight variation between the sequences. Yeast two-hybrid analysis demonstrated that TaKu70 and TaKu80 formed a heterodimer, and electrophoretic mobility shift assays revealed that this heterodimer bound to double-stranded DNA, but not to single-stranded DNA. The quantitative polymerase chain reaction analysis revealed that the expression of TaKu70 and TaKu80 genes was up-regulated under γ-ray irradiation in a dose-dependent manner in the seedlings of wheat. These results suggest that TaKu70 and TaKu80 form a functional heterodimer and are associated with the repair of the induced DSBs in wheat.

Keywords

DNA repairDSBKu proteinwheat
Type
Research Article
Information
Plant Genetic Resources , Volume 12 , Issue S1: Special issue on the 3rd International Symposium on “Genomics of Plant Genetic Resources” , July 2014 , pp. S99 - S103
Copyright
Copyright © NIAB 2014 

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