Interaction of the Warsaw breakage syndrome DNA helicase DDX11 with the replication fork-protection factor Timeless promotes sister chromatid cohesion.
Interaction of the Warsaw breakage syndrome DNA helicase DDX11 with the replication fork-protection factor Timeless promotes sister chromatid cohesion.
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DOI：
10.1371/journal.pgen.1007622
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发表时间：
2018-10
期刊：
影响因子：
4.5
通讯作者：
Pisani FM
中科院分区：
文献类型：
--
作者：
Cortone G;Zheng G;Pensieri P;Chiappetta V;Tatè R;Malacaria E;Pichierri P;Yu H;Pisani FM
姐妹染色单体凝聚力的建立与DNA复制有关，但其潜在的分子机制尚不完全清楚。DDX 11（也称为ChlR 1）是一种超家族2 Fe-S簇DNA解旋酶，与华沙断裂综合征（WABS）有关。在此，我们研究了DDX 11在人类细胞中的凝聚力建立中的作用。我们证明了DDX 11通过保守的肽基序与复制叉保护复合物的一个组成部分Timeless相互作用。DDX 11-Timeless相互作用对于间期和有丝分裂中的姐妹染色单体凝聚是至关重要的。免疫荧光研究进一步揭示了粘附素与染色质的结合需要DDX 11。最后，我们通过SIRF分析证明DDX 11定位于新生DNA。此外，我们发现DDX 11促进粘附素结合到DNA复制叉与Timeless一致，并且重组纯化的粘附素在体外与DDX 11相互作用。总的来说，我们的研究结果建立了一个关键的作用DDX 11-永恒的相互作用，在协调DNA复制与姐妹染色单体凝聚力，并有重要的影响，了解WABS的分子基础。染色体是包含遗传信息的DNA分子。在复制过程中，被蛋白质（姐妹染色单体）覆盖的两个姐妹DNA分子被称为cohesin的环状蛋白复合物的许多拷贝结合在一起，这一过程称为姐妹染色单体凝聚。在细胞分裂之前，两个姐妹染色单体的粘着环被移除，以允许它们向分裂母细胞的相反两极迁移。在这个过程结束时，两个子细胞继承了一套完整的染色体。在下一次细胞分裂之前，染色体以高速和保真度复制。这个重要的任务是由DNA复制机器完成的，这是一种由几种酶和蛋白质组成的复杂装置。在本研究中，我们已经证明，DDX 11和Timeless，两个亚基的DNA复制机器，招募的cohesin环，以促进其稳定的结合到新复制的染色体，即建立姐妹染色单体凝聚力。在通过基因工程降低DDX 11水平的人类细胞中，我们观察到姐妹染色单体凝聚力减弱，凝聚素与染色体的结合减少。我们的实验结果有助于我们理解的分子机制，在人类细胞中的DNA复制和姐妹染色单体凝聚力之间的功能耦合。
Establishment of sister chromatid cohesion is coupled to DNA replication, but the underlying molecular mechanisms are incompletely understood. DDX11 (also named ChlR1) is a super-family 2 Fe-S cluster-containing DNA helicase implicated in Warsaw breakage syndrome (WABS). Herein, we examined the role of DDX11 in cohesion establishment in human cells. We demonstrated that DDX11 interacts with Timeless, a component of the replication fork-protection complex, through a conserved peptide motif. The DDX11-Timeless interaction is critical for sister chromatid cohesion in interphase and mitosis. Immunofluorescence studies further revealed that cohesin association with chromatin requires DDX11. Finally, we demonstrated that DDX11 localises at nascent DNA by SIRF analysis. Moreover, we found that DDX11 promotes cohesin binding to the DNA replication forks in concert with Timeless and that recombinant purified cohesin interacts with DDX11 in vitro. Collectively, our results establish a critical role for the DDX11-Timeless interaction in coordinating DNA replication with sister chromatid cohesion, and have important implications for understanding the molecular basis of WABS. Chromosomes are DNA molecules that contain the genetic information. During replication, the two sister DNA molecules covered by proteins (sister chromatids) are held together by many copies of a ring-like protein complex named cohesin, in a process called sister-chromatid cohesion. Before a cell divides, the cohesin rings are removed from the two sister chromatids to allow their migration towards the opposite poles of the dividing mother cell. At the end of this process, the two daughter cells have inherited a complete set of chromosomes. Before the next cell division, chromosomes are duplicated with high speed and fidelity. This important task is performed by the DNA replication machinery, a sophisticated apparatus made of several enzymes and proteins. In the present study, we have demonstrated that DDX11 and Timeless, two subunits of the DNA replication machinery, recruit the cohesin rings to promote their stable binding to the newly duplicated chromosomes, that is the establishment of sister-chromatid cohesion. In human cells that were genetically engineered to reduce the level of DDX11, we observed that sister-chromatid cohesion was loosened and association of cohesin to chromosomes was reduced. Our experimental results contribute to our understanding of the molecular mechanisms underlying the functional coupling between DNA replication and sister-chromatid cohesion in human cells.
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影响因子：
19
作者：
Gligoris T;Löwe J
通讯作者：
Löwe J
DOI：
10.1007/s00018-014-1569-4
发表时间：
2014-07
期刊：
Cellular and molecular life sciences : CMLS
影响因子：
--
作者：
Bharti SK;Khan I;Banerjee T;Sommers JA;Wu Y;Brosh RM Jr
通讯作者：
Brosh RM Jr
影响因子：
--
作者：
Abe T;Kawasumi R;Arakawa H;Hori T;Shirahige K;Losada A;Fukagawa T;Branzei D
通讯作者：
Branzei D
影响因子：
4.8
作者：
Farina, Andrea;Shin, Jae-Ho;Hurwitz, Jerard
通讯作者：
Hurwitz, Jerard
影响因子：
4
作者：
Leman, Adam R.;Noguchi, Chiaki;Noguchi, Eishi
通讯作者：
Noguchi, Eishi