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Published November 1, 1988 | Published
Journal Article Open

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

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.

Additional Information

© 1988 National Academy of Sciences. Communicated by Earl W. Davie, July 25, 1988 (received for review April 3, 1988). We thank Drs. J. Ward, C. Young, and P. Christie for their critical reading of the manuscript. J. Ward, G. Bolton, and J. Shouguang supplied antisera. N.T. and A.D. were supported by Fulbright Fellowships. This work was supported by Grant 5 RO1 GM32618-14 from the National Institutes of Health. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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