Department of Life Sciences and Institute of Genome Sciences, National Yang-Ming University, Shih-Pai, Taipei 112, Taiwan.
Nucleic Acids Res. 2011 Mar;39(6):2165-74. doi: 10.1093/nar/gkq1204. Epub 2010 Nov 24.
Linear chromosomes and linear plasmids of Streptomyces possess covalently bound terminal proteins (TPs) at the 5' ends of their telomeres. These TPs are proposed to act as primers for DNA synthesis that patches the single-stranded gaps at the 3' ends during replication. Most ('archetypal') Streptomyces TPs (designated Tpg) are highly conserved in size and sequence. In addition, there are a number of atypical TPs with heterologous sequences and sizes, one of which is Tpc that caps SCP1 plasmid of Streptomyces coelicolor. Interactions between the TPs on the linear Streptomyces replicons have been suggested by electrophoretic behaviors of TP-capped DNA and circular genetic maps of Streptomyces chromosomes. Using chemical cross-linking, we demonstrated intramolecular and intermolecular interactions in vivo between Tpgs, between Tpcs and between Tpg and Tpc. Interactions between the chromosomal and plasmid telomeres were also detected in vivo. The intramolecular telomere interactions produced negative superhelicity in the linear DNA, which was relaxed by topoisomerase I. Such intramolecular association between the TPs poses a post-replicational complication in the formation of a pseudo-dimeric structure that requires resolution by exchanging TPs or DNA.
线性染色体和线性质粒的链霉菌拥有共价结合末端蛋白 (TPs) 在其端粒的 5' 端。这些 TPs 被提议作为引物用于 DNA 合成,在复制过程中修补 3' 端的单链缺口。大多数(“典型”)链霉菌 TPs(指定为 Tpg)在大小和序列上高度保守。此外,还有一些具有异源序列和大小的非典型 TPs,其中之一是 Tpc,它覆盖了链霉菌天蓝色素 SCP1 质粒。线性链霉菌复制子上的 TPs 之间的相互作用通过 TP 封端 DNA 的电泳行为和链霉菌染色体的圆形遗传图谱得到了暗示。使用化学交联,我们在体内证明了 Tpgs 之间、Tpcs 之间以及 Tpg 和 Tpc 之间的分子内和分子间相互作用。还在体内检测到染色体和质粒端粒之间的相互作用。分子内端粒相互作用在线性 DNA 中产生负超螺旋,拓扑异构酶 I 将其松弛。这种 TPs 之间的分子内缔合在形成需要通过交换 TPs 或 DNA 来解决的拟二聚体结构时会产生复制后并发症。