Linear Streptomyces plasmids form superhelical circles through interactions between their terminal proteins.
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.
Department of Life Sciences and Institute of Genome Sciences, National Yang-Ming University, Shih-Pai, Taipei 112, Taiwan.
Hsiu-Hui
HH
Tsai
Chih-Hung
CH
Huang
Ingrid
I
Tessmer
Dorothy A
DA
Erie
Carton W
CW
Chen
D001426:Bacterial Proteins / Q000378:metabolism*; D002875:Chromosomes / Q000378:metabolism; D003432:Cross-Linking Reagents; D004278:DNA, Superhelical / Q000648:ultrastructure*; D010957:Plasmids / Q000378:metabolism / Q000648:ultrastructure*; D013302:Streptomyces / Q000235:genetics* / Q000648:ultrastructure; D016615:Telomere / Q000378:metabolism; D034501:Telomere-Binding Proteins / Q000378:metabolism*
D001426:Bacterial Proteins; D003432:Cross-Linking Reagents; D004278:DNA, Superhelical; D034501:Telomere-Binding Proteins