Characterization of the gene Mre11 and evidence of silencing after polyploidization in Triticum

Abstract

The MRE11 protein is a component of the highly conserved MRN complex, along with RAD50 and NBS1. This complex is crucial in the repair of breaks in double stranded DNA, and is involved in many other cell processes. The present paper reports the molecular characterization of Mre11 gene in all three genomes of wheat, making use of the diploid species Triticum monococcum (genome A) and Aegilops Tauschii (genome D), the tetraploid T. turgidum (genomes A and B), and the hexaploid T. aestivum (genomes A, B and D). The genomic sequences characterized ranged from 4,662 to 4,766 bp in length; the cDNA corresponding to the processed mRNA was 2,440-2,510 bp long. In all cases, Mre11 coded for a highly conserved protein of 699 amino acids with a structure involving 22 exons. Mre11 expression was determined by real-time PCR in all the species analysed. The tetraploid species showed an expression similar to that of the diploid Ae. tauschii and lower than that of T. monococcum. Stronger expression was detected in the hexaploid T. aestivum. The SSCP technique was modified by introducing fluorescent labelling to the procedure in order to analyse the expression of the different Mre11 genes (i.e., those belonging to the different genomes) in the polyploid species. In both polyploids, the Mre11 gene belonging to the B genome was the least expressed. This probably reflects a first step in the process of silencing duplicate genes after polyploidization.