Analyses of DNA complex and crystallization of the DNA repair promoting protein PprA derived from Deinococcus Radiodurance

Adachi, Motoyasu; Tamada, Taro; Sato, Katsuya*; Narumi, Issei; Kuroki, Ryota

no journal, , 

The resistance of Deinococcus radiodurans to -ray is 1000 times higher than that of human cells. PprA is the novel protein which promotes repairing DNA, and play a key role in the ability to resist to the radiation. The characterization of PprA have showed: (1) that the expression of PprA is controlled under another novel protein of PprI (2) that PprA promotes ligase reaction: (3) that pprA exhibits the ability to bind on DNA. In this study, our objective is the crystal structure analysis of PprA to reveal its structure-function relationships and for application as genetic and clinical tools. Here, we constructed the E. coli expression and purification system of PprA. Using the purified PprA, some small crystals in size were obtained after screening of conditions for crystalization. Beased on the analyses of DNA-PprA interaction, we showed that 280 molecules of PprA at least can bind on a DNA molecule of pUC19 plasmid (2686bp). In addition, we indicated that several DNA-PprA complexes may form higher complex, when DNA includes terminal end or nick (the cleaved part on a DNA strand). These results are important knowledge in molecular assembly and function of PprA.