The RAD51 gene family in bread wheat is highly conserved across eukaryotes, with RAD51A upregulated during early meiosis
Kelvin H. P. Khoo A , Hayley R. Jolly A and Jason A. Able A BA School of Agriculture, Food and Wine, The University of Adelaide, Waite Campus, PMB1, Glen Osmond, SA 5064, Australia.
B Corresponding author. Email: jason.able@adelaide.edu.au
Functional Plant Biology 35(12) 1267-1277 https://doi.org/10.1071/FP08203
Submitted: 21 July 2008 Accepted: 25 September 2008 Published: 16 December 2008
Abstract
The RADiation sensitive protein 51 (RAD51) recombinase is a eukaryotic homologue of the bacterial Recombinase A (RecA). It is required for homologous recombination of DNA during meiosis where it plays a role in processes such as homology searching and strand invasion. RAD51 is well conserved in eukaryotes with as many as four paralogues identified in vertebrates and some higher plants. Here we report the isolation and preliminary characterisation of four RAD51 gene family members in hexaploid (bread) wheat (Triticum aestivum L.). RAD51A1, RAD51A2 and RAD51D were located on chromosome group 7, and RAD51C was on chromosome group 2. Q-PCR gene expression profiling revealed that RAD51A1 was upregulated during meiosis with lower expression levels seen in mitotic tissue, and bioinformatics analysis demonstrated the evolutionary linkages of this gene family to other eukaryotic RAD51 sequences. Western blot analysis of heterologously expressed RAD51 from bread wheat has shown that it is detectable using anti-human RAD51 antibodies and that molecular modelling of the same protein revealed structural conservation when compared with yeast, human, Arabidopsis and maize RAD51A orthologues. This report has widened the knowledge base of this important protein family in plants, and highlighted the high level of structural conservation among RAD51 proteins from various species.
Additional keywords: evolution, recombination.
Acknowledgements
The authors thank Professor Akira Shinohara (Faculty of Science, Osaka University, Japan) for supplying the anti-HsRAD51 antibody used in this study, and Dr Amanda J Able (School of Agriculture, Food and Wine, The University of Adelaide) for reviewing the manuscript. This research was supported in part by the Molecular Plant Breeding Cooperative Research Centre (MPB CRC), the Australian Government under the Australia-India Strategic Research Fund (AISRF) and the School of Agriculture, Food and Wine, The University of Adelaide.
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