Smith Matthew C A, Thomas Christopher D
Astbury Centre for Structural Molecular Biology, School of Biochemistry and Molecular Biology, University of Leeds, Leeds LS2 9JT, United Kingdom.
J Bacteriol. 2004 Jun;186(11):3363-73. doi: 10.1128/JB.186.11.3363-3373.2004.
Mobilization of the staphylococcal plasmid pC221 requires at least one plasmid-encoded protein, MobA, in order to form a relaxosome. pC221 and closely related plasmids also possess an overlapping reading frame encoding a protein of 15 kDa, termed MobC. By completing the nucleotide sequence of plasmid pC223, we have found a further example of this small protein, and gene knockouts have shown that MobC is essential for relaxosome formation and plasmid mobilization in both pC221 and pC223. Primer extension analysis has been used to identify the nic site in both of these plasmids, located upstream of the mobC gene in the sense strand. Although the sequence surrounding the nic site is highly conserved between pC221 and pC223, exchange of the oriT sequence between plasmids significantly reduces the extent of relaxation complex formation, suggesting that the Mob proteins are selective for their cognate plasmids in vivo.
葡萄球菌质粒pC221的迁移需要至少一种质粒编码蛋白MobA,以形成松弛体。pC221及密切相关的质粒还拥有一个重叠阅读框,编码一种15 kDa的蛋白,称为MobC。通过完成质粒pC223的核苷酸序列测定,我们发现了这种小蛋白的又一个实例,基因敲除实验表明,MobC对于pC221和pC223中松弛体的形成及质粒迁移至关重要。引物延伸分析已用于确定这两种质粒中的nic位点,该位点位于正义链上mobC基因的上游。虽然pC221和pC223之间nic位点周围的序列高度保守,但质粒之间oriT序列的交换显著降低了松弛复合体形成的程度,这表明Mob蛋白在体内对其同源质粒具有选择性。