Pif1 is essential for efficient replisome progression through lagging strand G-quadruplex DNA secondary structures.
Pif1 is essential for efficient replisome progression through lagging strand G-quadruplex DNA secondary structures.
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DOI：
10.1093/nar/gky1065
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发表时间：
2018-12-14
影响因子：
14.9
通讯作者：
Aharoni A
中科院分区：
文献类型：
--
作者：
Dahan D;Tsirkas I;Dovrat D;Sparks MA;Singh SP;Galletto R;Aharoni A
Pif1 DNA解旋酶是体外g -四重体（G4）结构的有效解旋酶，其功能是维持酿酒酵母G4序列的基因组稳定性。在这里，我们开发并利用了一种活细胞成像方法，通过单个酵母细胞中不同的含G4序列来定量测量单个复制叉的进展速率。我们发现在缺乏Pif1的情况下，通过特定滞后链G4序列在体内的复制率显著降低。相反，我们发现在缺乏Pif1的情况下，通过前导链上相同的G4s的复制速率相对于各自的WT菌株并没有降低，这表明Pif1仅对通过后导链G4s的有效复制至关重要。此外，我们发现Pif1中的典型PIP序列与PCNA相互作用，并且在体外和体内缺乏这种相互作用时，通过G4结构的复制明显减慢。因此，Pif1 - pcna相互作用对于通过G4序列优化复制体进展至关重要，突出了酵母基因组复制过程中Pif1活性与复制体进展之间耦合的重要性。
Pif1 DNA helicase is a potent unwinder of G-quadruplex (G4) structures in vitro and functions to maintain genome stability at G4 sequences in Saccharomyces cerevisiae. Here, we developed and utilized a live-cell imaging approach to quantitatively measure the progression rates of single replication forks through different G4 containing sequences in individual yeast cells. We show that in the absence of Pif1, replication rates through specific lagging strand G4 sequences in vivo is significantly decreased. In contrast, we found that in the absence of Pif1, replication rates through the same G4s on the leading strand are not decreased relative to the respective WT strains, showing that Pif1 is essential only for efficient replication through lagging strand G4s. Additionally, we show that a canonical PIP sequence in Pif1 interacts with PCNA and that replication through G4 structures is significantly slower in the absence of this interaction in vitro and in vivo. Thus, Pif1–PCNA interaction is essential for optimal replisome progression through G4 sequences, highlighting the importance of coupling between Pif1 activity and replisome progression during yeast genome replication.
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影响因子：
3.5
作者：
Leman AR;Noguchi E
通讯作者：
Noguchi E
DOI：
10.1038/nrm.2016.43
发表时间：
2016-06
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
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通讯作者：
Sfeir A
影响因子：
16
作者：
Haracska, L;Kondratick, CM;Prakash, L
通讯作者：
Prakash, L
DOI：
10.1038/nrg3296
发表时间：
2012-11
期刊：
Nature reviews. Genetics
影响因子：
--
作者：
通讯作者：
--
影响因子：
3.8
作者：
Bochman ML;Sabouri N;Zakian VA
通讯作者：
Zakian VA