Ziegelin G, Fürste J P, Lanka E
Max-Planck-Institut für Molekulare Genetik, Abteilung Schuster, Berlin, West Germany.
J Biol Chem. 1989 Jul 15;264(20):11989-94.
Transfer of plasmid RP4 during bacterial conjugation requires the plasmid-encoded TraJ protein, which binds to the transfer origin (Fürste, J. P., Pansegrau, W., Ziegelin, G., Kröger, M., and Lanka, E. (1989) Proc. Natl. Acad. Sci. U.S.A. 86, 1771-1775). As indicated by traJ mutants, the TraJ protein is a constituent of the relaxosome, the initiation complex of transfer DNA replication. The traJ gene maps adjacent to the transfer origin (oriT). The structural gene consists of a 372-base pair sequence encoding a polypeptide of 122 amino acids (13,282 Da). TraJ was purified from an Escherichia coli strain overproducing the protein. DNA footprinting experiments involving DNase I demonstrated that the purified protein binds to the right arm of a 19-base pair inverted repeat within oriT. Hydroxyl radical footprints of the DNA-protein complex revealed that TraJ protein is bound to only one side of the DNA helix.
在细菌接合过程中,质粒RP4的转移需要质粒编码的TraJ蛋白,该蛋白与转移起始点结合(Fürste, J. P., Pansegrau, W., Ziegelin, G., Kröger, M., and Lanka, E. (1989) Proc. Natl. Acad. Sci. U.S.A. 86, 1771 - 1775)。如traJ突变体所示,TraJ蛋白是松弛体的组成部分,松弛体是转移DNA复制的起始复合物。traJ基因定位于与转移起始点(oriT)相邻的位置。该结构基因由一个372个碱基对的序列组成,编码一个122个氨基酸(13,282道尔顿)的多肽。TraJ是从过量表达该蛋白的大肠杆菌菌株中纯化得到的。涉及DNase I的DNA足迹实验表明,纯化后的蛋白与oriT内一个19个碱基对反向重复序列的右臂结合。DNA - 蛋白复合物的羟基自由基足迹显示,TraJ蛋白仅与DNA螺旋的一侧结合。