Graduate Thesis Or Dissertation
 

Functional domains of Agrobacterium tumefaciens single-stranded DNA binding protein VirE2

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  • Agrobacterium tumefaciens is a gram-negative soil bacterium that causes crown gall tumors on dicotyledenous plants. The transferred DNA (T-DNA) portion of the A. tumefaciens tumor-inducing (Ti) plasmid enters infected plant cells and integrates into plant nuclear DNA. The T-DNA is accompanied into plant cells by the VirD2 endonuclease covalently attached to its 5' end. VirE2, a cooperative, single-stranded DNA-binding protein is also transported into plant cells during infection by A. tumefaciens. VirD2 and VirE2 contain nuclear localization signals (NLSs) and are transported into the plant cell nucleus. The location of functional domains by the insertion of Xhol linker oligonucleotides throughout virE2 is reported. A ssDNA binding domain was located in the C-terminal half of VirE2, as well two domains involved in cooperative single-stranded DNA binding. Further, we isolated a mutation in the central region of VirE2 that decreased tumorigenicity, but did not affect ssDNA binding.
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