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-Structure paper
タイトル | Ctf18-RFC and DNA Pol ϵ form a stable leading strand polymerase/clamp loader complex required for normal and perturbed DNA replication. |
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ジャーナル・号・ページ | Nucleic Acids Res, Vol. 48, Issue 14, Page 8128-8145, Year 2020 |
掲載日 | 2020年8月20日 |
![]() | Katy Stokes / Alicja Winczura / Boyuan Song / Giacomo De Piccoli / Daniel B Grabarczyk / ![]() ![]() |
PubMed 要旨 | The eukaryotic replisome must faithfully replicate DNA and cope with replication fork blocks and stalling, while simultaneously promoting sister chromatid cohesion. Ctf18-RFC is an alternative PCNA ...The eukaryotic replisome must faithfully replicate DNA and cope with replication fork blocks and stalling, while simultaneously promoting sister chromatid cohesion. Ctf18-RFC is an alternative PCNA loader that links all these processes together by an unknown mechanism. Here, we use integrative structural biology combined with yeast genetics and biochemistry to highlight the specific functions that Ctf18-RFC plays within the leading strand machinery via an interaction with the catalytic domain of DNA Pol ϵ. We show that a large and unusually flexible interface enables this interaction to occur constitutively throughout the cell cycle and regardless of whether forks are replicating or stalled. We reveal that, by being anchored to the leading strand polymerase, Ctf18-RFC can rapidly signal fork stalling to activate the S phase checkpoint. Moreover, we demonstrate that, independently of checkpoint signaling or chromosome cohesion, Ctf18-RFC functions in parallel to Chl1 and Mrc1 to protect replication forks and cell viability. |
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手法 | EM (単粒子) / X線回折 |
解像度 | 4.2 - 6.1 Å |
構造データ | EMDB-10088: Ctf18-1-8 in complex with the catalytic domain of DNA polymerase epsilon EMDB-10089, PDB-6s2f: ![]() PDB-6s1c: |
化合物 | ![]() ChemComp-SF4: |
由来 |
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![]() | REPLICATION / DNA polymerase / PCNA loader / protein complex |