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. 1988 Nov;85(22):8558–8562. doi: 10.1073/pnas.85.22.8558

Role of the overdrive sequence in T-DNA border cleavage in Agrobacterium.

N Toro 1, A Datta 1, M Yanofsky 1, E Nester 1
PMCID: PMC282498  PMID: 3186745

Abstract

The T-DNA of the Ti plasmid of Agrobacterium is flanked by 25-base-pair imperfect direct repeats that are required in cis for transfer to the genome of the plant host. Another sequence, designated overdrive, is located adjacent to the right-border repeats and functions in cis to enhance tumor formation. We have examined the effect of the overdrive sequence on the early steps in T-DNA processing. We report here that overdrive greatly enhances cleavage by the site-specific endonuclease in Agrobacterium, perhaps by directing the endonuclease to the adjacent border sequences. We also show by a gel mobility-shift assay that overdrive affinity-purified proteins from acetosyringone-induced Agrobacterium cells interact with T-DNA border and overdrive sequences. Further, we show that in vivo the virC operon enhances cleavage at the T-DNA borders, most likely by interaction between the VirC1 protein and the overdrive sequence.

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