Determining the structure of the Deinococcus radiodurans nucleoid-associated protein DR0199 by X-ray crystallography
Meggan A Bridgett
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Nucleoid-associated proteins (NAPs) are important for binding, compacting, and organizing bacterial DNA housed in the nucleoid. These proteins are present in numerous bacterial organisms including Deinococcus radiodurans. This organism is closely studied for its unique ability to survive after exposure to DNA damaging conditions. It is believed that NAPs are an important component in the ability to repair DNA after it has been damaged. This study focused on determining the structure of the NAP DR0199 found in Deinococcus radiodurans through protein overexpression, purification, crystallization, and X-ray crystallography. DR0199 crystals achieved successful growth, however the highest resolution of the X-ray crystallography data was 4 Å which was not a high enough resolution for a probable structure to be determined. This study further examined the relationship between the DR0199 NAP and other proteins expressed in the Deinococcus radiodurans organism. Deinococcus radiodurans cultures were exposed to DNA stressing conditions to encourage expression of proteins most likely to function with DR0199. The cells were then lysed and run through a column containing the DR0199 protein, so that proteins whose function allows them to physically interact with DR0199 would bind to the DR0199 protein in the column. Bound proteins were eluted and analyzed by SDS-PAGE.
Honors Thesis - Undergraduate
Matthew E. Lopper
Primary Advisor's Department
Stander Symposium project
"Determining the structure of the Deinococcus radiodurans nucleoid-associated protein DR0199 by X-ray crystallography" (2017). Stander Symposium Projects. 1079.