期刊
NATURE STRUCTURAL & MOLECULAR BIOLOGY
卷 22, 期 12, 页码 976-982出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nsmb.3113
关键词
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资金
- US National Institutes of Health [GM103314, GM109824, GM74985, AG29979, GM38839]
- Howard Hughes Medical Institute
At the eukaryotic DNA replication fork, it is widely believed that the Cdc45-Mcm2-7-GINS (CMG) helicase is positioned in front to unwind DNA and that DNA polymerases trail behind the helicase. Here we used single-particle EM to directly image a Saccharomyces cerevisiae replisome. Contrary to expectations, the leading strand Pol epsilon is positioned ahead of CMG helicase, whereas Ctf4 and the lagging-strand polymerase (Pol) alpha-primase are behind the helicase. This unexpected architecture indicates that the leading-strand DNA travels a long distance before reaching Pol epsilon, first threading through the Mcm2-7 ring and then making a U-turn at the bottom and reaching Pol a at the top of CMG. Our work reveals an unexpected configuration of the eukaryotic replisome, suggests possible reasons for this architecture and provides a basis for further structural and biochemical replisome studies.
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