Huang Chih-Hung, Tsai Hsiu-Hui, Tsay Yeou-Guang, Chien Yu-Nien, Wang Shan-Li, Cheng Ming-Yuan, Ke Chia-Hui, Chen Carton W
Institute of Biotechnology, National Taipei University of Technology, Taipei 106, Taiwan.
Mol Microbiol. 2007 Mar;63(6):1710-8. doi: 10.1111/j.1365-2958.2007.05616.x.
Linear plasmids and chromosomes of Streptomyces carry terminal proteins (TPs) covalently attached to the 5' ends of the DNA. Most known telomeres are conserved in primary sequence and in the potential secondary structures formed during replication. The TP that caps these telomeres is also highly conserved and its coding gene, tpg, is present in all Streptomyces chromosomes and some linear plasmids. Linear plasmid SCP1 contains atypical telomere sequences and no tpg homologue, and can replicate in the absence of tpg, suggesting that it carries a novel TP gene. To isolate the TP on the SCP1 telomeres, we constructed a multicopy mini-SCP1 plasmid. The TP capping the plasmid was isolated and subjected to tryptic digestion and mass spectrometric analysis, and the results indicated that the TP was encoded by an open reading frame (ORF), SCP1.127 (tpc), on SCP1. Of the two ORFs upstream of tpc, SCP1.125 (tac) but not SCP1.126 was essential for replication of mini-SCP1. The Tac-Tpc system of SCP1 represents a convergently evolved novel telomere-capping system of Streptomyces linear replicons.
链霉菌的线性质粒和染色体携带与DNA 5'端共价连接的末端蛋白(TPs)。大多数已知的端粒在一级序列以及复制过程中形成的潜在二级结构方面都是保守的。封端这些端粒的TP也是高度保守的,其编码基因tpg存在于所有链霉菌染色体和一些线性质粒中。线性质粒SCP1含有非典型的端粒序列且没有tpg同源物,并且可以在没有tpg的情况下复制,这表明它携带一个新的TP基因。为了分离SCP1端粒上的TP,我们构建了一个多拷贝的mini-SCP1质粒。分离出封端该质粒的TP并进行胰蛋白酶消化和质谱分析,结果表明该TP由SCP1上的一个开放阅读框(ORF)SCP1.127(tpc)编码。在tpc上游的两个ORF中,SCP1.125(tac)而非SCP1.126对mini-SCP1的复制至关重要。SCP1的Tac-Tpc系统代表了链霉菌线性复制子中一个趋同进化的新型端粒封端系统。