In vivo Recombination After Chronic Damage Exposure Falls to Below Spontaneous Levels in ‘.pdf

In vivo Recombination After Chronic Damage Exposure Falls to Below Spontaneous Levels in ‘‘Recombomice’’ Olga Kovalchuk,1 Carrie A. Hendricks,2 Scott Cassie,1 Andrew J. Engelward,3 and Bevin P. Engelward2 1Department of Biological Sciences, University of Lethbridge, Lethbridge, Alberta, Canada; 2Biological Engineering Division, Massachusetts Institute of Technology, Cambridge, Massachusetts; and 3Department of Mathematics, Harvard University, Cambridge, Massachusetts Abstract All forms of cancer are initiated by heritable changes in gene expression. Although point mutations have been studied extensively, much less is known about homologous recombination events, despite its role in causing sequence rearrangements that contribute to tumorigenesis. Although transgenic mice that permit detection of point mutations have provided a fundamental tool for studying point mutations in vivo, until recently, transgenic mice designed specifically to detect homologous recombination events in somatic tissues in vivo did not exist. We therefore created fluorescent yellow direct repeat mice, enabling automated detection of recombinant cells in vivo for the first time. Here, we show that an acute dose of ionizing radiation induces recombination in fluorescent yellow direct repeat mice, providing some of the first direct evidence that ionizing radiation induces homologous recombination in cutaneous tissues in vivo. In contrast, the same total dose of radiation given under chronic exposure conditions suppresses recombination to levels that are significantly below those of unexposed animals. In addition, global methylation is suppressed and key DNA repair proteins are induced in tissues from chronically irradiated animals (specifically AP endonuclease, polymerase B, and Ku70). Thus, increased clearance of recombinogenic lesions may contribute to suppression of homologous recombination. Taken together, these studies show that fluorescent yellow direct repeat mice provide a rapid and power