Prolyl Isomerization-Mediated Conformational Changes Define ATR Subcellular Compartment-Specific Functions.
Prolyl Isomerization-Mediated Conformational Changes Define ATR Subcellular Compartment-Specific Functions.
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DOI：
10.3389/fcell.2022.826576
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发表时间：
2022
影响因子：
5.5
通讯作者：
中科院分区：
文献类型：
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ATR是一种PI3K样激酶蛋白，调节对DNA损伤和复制应激的检查点反应。除了其在细胞核中的检查点功能外，ATR在DNA损伤后在线粒体中积极参与抗凋亡作用。ATR在细胞核和细胞质中的不同功能分别由ATR的两种脯氨酰异构体形式：反式和顺式ATR执行。该异构化发生在ATR的Pin1 Ser428-Pro429基序处。在这里，我们研究了人类ATR的亚细胞位置特异性功能的结构基础。使用基于质谱的足迹法，ATR赖氨酸残基的磺基-NHS-LC-生物素修饰的表面可及性进行监测，并在顺式和反式异构体之间进行比较。我们已经确定了两个生物素修饰的赖氨酸残基，K459和K469，在顺式ATR的BH3样结构域内，在反式ATR中不可及，表明顺式和反式ATR之间围绕BH3结构域的构象变化。构象变化还涉及N-末端结构域和中间HEAT结构域。此外，从一系列互补测定的实验结果表明，具有可接近的BH 3结构域的顺式ATR能够结合tBid，而trans-ATR不能。此外，顺式和反式ATR可以直接形成同源二聚体通过其C-末端结构域没有ATRIP，而核（trans-ATR）在ATRIP的存在下形成二聚体-二聚体复合物涉及的N-和C-末端的ATR和ATRIP后UV。通过分子模拟和动力学模拟分析了Ser428-Pro429基序和BH3结构域区域的结构特征。在支持，顺式构象被发现是显着更积极有利的比反式在Ser428-Pro429键在20个氨基酸的野生型ATR肽。两者合计，我们的研究结果表明，异构化诱导的ATR结构变化定义其亚细胞位置和隔室特异性功能，并在促进细胞存活和DNA损伤反应中发挥重要作用。
ATR is a PI3K-like kinase protein, regulating checkpoint responses to DNA damage and replication stress. Apart from its checkpoint function in the nucleus, ATR actively engages in an antiapoptotic role at mitochondria following DNA damage. The different functions of ATR in the nucleus and cytoplasm are carried out by two prolyl isomeric forms of ATR: trans- and cis-ATR, respectively. The isomerization occurs at the Pin1 Ser428-Pro429 motif of ATR. Here, we investigated the structural basis of the subcellular location-specific functions of human ATR. Using a mass spectrometry-based footprinting approach, the surface accessibility of ATR lysine residues to sulfo-NHS-LC-biotin modification was monitored and compared between the cis- and the trans-isomers. We have identified two biotin-modified lysine residues, K459 and K469, within the BH3-like domain of cis-ATR that were not accessible in trans-ATR, indicating a conformational change around the BH3 domain between cis- and trans-ATR. The conformational alteration also involved the N-terminal domain and the middle HEAT domain. Moreover, experimental results from an array of complementary assays show that cis-ATR with the accessible BH3 domain was able to bind to tBid while trans-ATR could not. In addition, both cis- and trans-ATR can directly form homodimers via their C-terminal domains without ATRIP, while nuclear (trans-ATR) in the presence of ATRIP forms dimer–dimer complexes involving both N- and C-termini of ATR and ATRIP after UV. Structural characteristics around the Ser428-Pro429 motif and the BH3 domain region are also analyzed by molecular modeling and dynamics simulation. In support, cis conformation was found to be significantly more energetically favorable than trans at the Ser428-Pro429 bond in a 20-aa wild-type ATR peptide. Taken together, our results suggest that the isomerization-induced structural changes of ATR define both its subcellular location and compartment-specific functions and play an essential role in promoting cell survival and DNA damage responses.
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影响因子：
16
作者：
Liu S;Shiotani B;Lahiri M;Maréchal A;Tse A;Leung CC;Glover JN;Yang XH;Zou L
通讯作者：
Zou L
ATR在线粒体上扮演直接的抗凋亡作用，该抗凋亡作用受Prolyl异构酶PIN1的调节。
影响因子：
16
作者：
Hilton BA;Li Z;Musich PR;Wang H;Cartwright BM;Serrano M;Zhou XZ;Lu KP;Zou Y
通讯作者：
Zou Y
影响因子：
14.9
作者：
Perez-Riverol Y;Bai J;Bandla C;García-Seisdedos D;Hewapathirana S;Kamatchinathan S;Kundu DJ;Prakash A;Frericks-Zipper A;Eisenacher M;Walzer M;Wang S;Brazma A;Vizcaíno JA
通讯作者：
Vizcaíno JA
影响因子：
10.5
作者：
Mordes, Daniel A.;Glick, Gloria G.;Cortez, David
通讯作者：
Cortez, David
DOI：
10.1385/1-59259-045-4:113
发表时间：
2000-01-01
期刊：
MASS SPECTROMETRY OF PROTEINS AND PEPTIDES
影响因子：
--
作者：
Bennett, KL;Matthiesen, T;Roepstorff, P
通讯作者：
Roepstorff, P