COVALENTLY BOUND VIRD2 PROTEIN OF AGROBACTERIUM-TUMEFACIENS PROTECTS THE T-DNA FROM EXONUCLEOLYTIC DEGRADATION
COVALENTLY BOUND VIRD2 PROTEIN OF AGROBACTERIUM-TUMEFACIENS PROTECTS THE T-DNA FROM EXONUCLEOLYTIC DEGRADATION
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DOI：
10.1073/pnas.86.23.9154
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发表时间：
1989-12-01
影响因子：
11.1
通讯作者：
KOUKOLIKOVANICOLA, Z
中科院分区：
文献类型：
--
作者：
DURRENBERGER, F;CRAMERI, A;KOUKOLIKOVANICOLA, Z
我们发现，在农杆菌的诱导下，自由的线性双链T-DNA分子以及之前描述的t链从Ti质粒中产生。这些分子中的大多数都与蛋白质结合在一起。我们发现这种蛋白是毒力基因virD2的产物。该蛋白被发现附着在加工过的T-DNA的5'末端的右侧边界和Ti质粒的左侧边界上。Pronase消化后的蛋白残余物使双链T-DNA右端对5'' .fwdarw具有抗性。体外3‘’外核溶解攻击。蛋白质- dna结合对SDS、巯基乙醇、温和碱、哌啶和羟胺具有抗性，表明它涉及共价键。讨论了这种t -DNA-蛋白复合物在复制、转导到植物、核靶向和整合到植物核DNA中的可能参与。
We show that upon induction of Agrobacterium tumefaciens, free linear double-stranded T-DNA molecules as well as the previously described T-strands are generated from the Ti plasmid. A majority of these molecules are bound to a protein. We show that this protein is the product of the virulence gene virD2. This protein was found to be attached to the 5'' terminus of processed T-DNA at the right border and to the rest of the Ti plasmid at the left border. The protein remnant after Pronase digestion rendered the right end of the doulbe-stranded T-DNA resistant to 5'' .fwdarw. 3'' exonucleolytic attack in vitro. The protein-DNA association was resistant to SDS, mercaptoethanol, mild alkali, piperidine, and hydroxylamine, indicating that it involves a covalent linkage. The possible involvement of this T-DNA-protein complex in replication, transduction to the plant, nuclear targeting, and integration into the plant nuclear DNA is discussed.