Agrobacterium May Delay Plant Nonhomologous End-Joining DNA Repair via XRCC4 to Favor T-DNA Integration

Agrobacterium tumefaciens is a soilborne pathogen that causes crown gall disease in many dicotyledonous plants by transfer of a portion of its tumor-inducing plasmid (T-DNA) into the plant genome. Several plant factors that play a role in Agrobacterium attachment to plant cells and transport of T-DN...

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Published in:	The Plant cell 2012-10, Vol.24 (10), p.4110-4123
Main Authors:	Vaghchhipawala, Zarir E., Vasudevan, Balaji, Lee, Seonghee, Morsy, Mustafa R., Mysore, Kirankumar S.
Format:	Article
Language:	eng
Subjects:	Agrobacterium Agrobacterium tumefaciens Agrobacterium tumefaciens - genetics Arabidopsis - genetics Arabidopsis - metabolism Arabidopsis - microbiology Arabidopsis Proteins - analysis Arabidopsis Proteins - metabolism Arabidopsis Proteins - physiology Arabidopsis thaliana Bacterial Proteins - metabolism Bleomycin - pharmacology Comets Diseases and pests DNA DNA Breaks, Double-Stranded DNA damage DNA End-Joining Repair DNA Repair DNA, Bacterial - metabolism DNA-Binding Proteins - analysis DNA-Binding Proteins - metabolism DNA-Binding Proteins - physiology Down-Regulation Gene Silencing Genetic aspects Genome, Plant Genomics Health aspects Host-parasite relationships Ion Channels - metabolism Molecular Sequence Data Nicotiana - genetics Nicotiana - metabolism Nicotiana - microbiology Physiological aspects Plant cells Plant genetics Plant Proteins - analysis Plant Proteins - metabolism Plant Proteins - physiology Plant roots Plants Transformation, Genetic Tumors
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Summary:	Agrobacterium tumefaciens is a soilborne pathogen that causes crown gall disease in many dicotyledonous plants by transfer of a portion of its tumor-inducing plasmid (T-DNA) into the plant genome. Several plant factors that play a role in Agrobacterium attachment to plant cells and transport of T-DNA to the nucleus have been identified, but the T-DNA integration step during transformation is poorly understood and has been proposed to occur via nonhomologous end-joining (NHEJ)—mediated double-strand DNA break (DSB) repair. Here, we report a negative role of X-RAY CROSS COMPLEMENTATION GROUP4 (XRCC4), one of the key proteins required for NHEJ, in Agrobacterium T-DNA integration. Downregulation of XRCC4 in Arabidopsis and Nicotiana benthamiana increased stable transformation due to increased T-DNA integration. Overexpression of XRCC4 in Arabidopsis decreased stable transformation due to decreased T-DNA integration. Interestingly, XRCC4 directly interacted with Agrobacterium protein VirE2 in a yeast two-hybrid system and in planta. VirE2-expressing Arabidopsis plants were more susceptible to the DNA damaging chemical bleomycin and showed increased stable transformation. We hypothesize that VirE2 titrates or excludes active XRCC4 protein available for DSB repair, thus delaying the closure of DSBs in the chromosome, providing greater opportunity for T-DNA to integrate.
ISSN:	1040-4651 1532-298X