Niwa Y, Yamamoto A, Machida C, Takebe I, Machida Y
Department of Biology, Faculty of Science, Nagoya University, Japan.
Nucleic Acids Res. 1988 Aug 11;16(15):7647-61. doi: 10.1093/nar/16.15.7647.
The T-DNA of octopine Ti plasmid of Agrobacterium tumefaciens contains TL- and TR-DNA regions each bounded by 25 base-pair-repeats (designated A, B, C and D from left to right). Short DNA segments containing the borders B, C and D were found to function as promoter when placed in the rightward orientation upstream of promoter-less lacZ. Promoter consensus sequence of Agrobacterium were found within these border repeats and in their adjacent regions. The expression of lacZ was low when the segments contained the overdrive, a sequence known to enhance T-DNA transfer. Simultaneous overproduction of VirD1 and D2 proteins, endonuclease acting on the border repeats, interfered with the promoter functions of the border segments. In spite of their activity under these conditions, the border regions do not seem to be involved in the gene expression, because they are not followed by appropriate open reading frames. We propose that RNA polymerase of Agrobacterium competes with VirD products for T-DNA borders and thereby affects the transfer of T-DNA.
根癌农杆菌章鱼碱型Ti质粒的T-DNA包含TL-DNA和TR-DNA区域,每个区域都由25个碱基对的重复序列界定(从左到右分别命名为A、B、C和D)。发现含有边界B、C和D的短DNA片段,当以向右的方向置于无启动子的lacZ上游时,可作为启动子发挥作用。在这些边界重复序列及其相邻区域发现了农杆菌的启动子共有序列。当片段包含增强子序列(一种已知可增强T-DNA转移的序列)时,lacZ的表达较低。同时过量产生作用于边界重复序列的核酸内切酶VirD1和D2蛋白,会干扰边界片段的启动子功能。尽管在这些条件下它们具有活性,但边界区域似乎不参与基因表达,因为它们后面没有合适的开放阅读框。我们提出,农杆菌的RNA聚合酶与VirD产物竞争T-DNA边界,从而影响T-DNA的转移。