Yang Chien-Chin, Tseng Shu-Min, Chen Carton W
Department of Chemistry, Chung-Yuan Christian University, Chung-li 32023, Taiwan
Department of Chemistry, Chung-Yuan Christian University, Chung-li 32023, Taiwan.
Nucleic Acids Res. 2015 Jul 27;43(13):6373-83. doi: 10.1093/nar/gkv302. Epub 2015 Apr 16.
Typical telomeres of linear chromosomes and plasmids of soil bacteria Streptomyces consist of tightly packed palindromic sequences with a terminal protein ('TP') covalently attached to the 5' end of the DNA. Replication of these linear replicons is initiated internally and proceeds bidirectionally toward the telomeres, which leaves single-strand overhangs at the 3' ends. These overhangs are filled by DNA synthesis using the TPs as the primers ('end patching'). The gene encoding for typical TP, tpg, forms an operon with tap, encoding an essential telomere-associated protein, which binds TP and the secondary structures formed by the 3' overhangs. Previously one of the two translesion synthesis DNA polymerases, DinB1 or DinB2, was proposed to catalyze the protein-primed synthesis. However, using an in vitro end-patching system, we discovered that Tpg and Tap alone could carry out the protein-primed synthesis to a length of 13 nt. Similarly, an 'atypical' terminal protein, Tpc, and its cognate telomere-associated protein, Tac, of SCP1 plasmid, were sufficient to achieve protein-primed synthesis in the absence of additional polymerase. These results indicate that these two telomere-associated proteins possess polymerase activities alone or in complex with the cognate TPs.
土壤细菌链霉菌的线性染色体和质粒的典型端粒由紧密堆积的回文序列组成,其末端有一个末端蛋白(“TP”)共价连接到DNA的5'端。这些线性复制子的复制在内部起始,并向端粒双向进行,这在3'端留下单链突出端。这些突出端通过以TP为引物的DNA合成来填充(“末端修补”)。编码典型TP的基因tpg与tap形成一个操纵子,tap编码一种必需的端粒相关蛋白(Tap),它结合TP以及由3'突出端形成的二级结构。此前有人提出两种跨损伤合成DNA聚合酶之一,DinB1或DinB2,催化蛋白引发的合成。然而,使用体外末端修补系统,我们发现单独的Tpg和Tap能够进行长达13个核苷酸的蛋白引发合成。同样,SCP1质粒的“非典型”末端蛋白Tpc及其同源端粒相关蛋白Tac,在没有额外聚合酶的情况下也足以实现蛋白引发合成。这些结果表明,这两种端粒相关蛋白单独或与同源TP形成复合物时都具有聚合酶活性。