Abstract
ATR (ataxia telangiectasia-mutated and Rad3-related) protein kinase and ATRIP (ATR-interacting protein) form a complex and play a critical role in response to replication stress and DNA damage. Here, we determined the cryo-electron microscopy (EM) structure of the human ATR-ATRIP complex at 4.7 Å resolution and built an atomic model of the C-terminal catalytic core of ATR (residues 1 521-2 644) at 3.9 Å resolution. The complex adopts a hollow “heart” shape, consisting of two ATR monomers in distinct conformations. The EM map for ATRIP reveals 14 HEAT repeats in an extended “S” shape. The conformational flexibility of ATR allows ATRIP to properly lock the N-termini of the two ATR monomers to favor ATR-ATRIP complex formation and functional diversity. The isolated “head-head” and “tail-tail” each adopts a pseudo 2-fold symmetry. The catalytic pockets face outward and substrate access is not restricted by inhibitory elements. Our studies provide a structural basis for understanding the assembly of the ATR-ATRIP complex and a framework for characterizing ATR-mediated DNA repair pathways.
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Acknowledgements
We thank the National Center for Protein Science Shanghai, Tsinghua University Branch of the National Protein Science Facility (Beijing), Center for Biological Imaging of Institute of Biophysics of the Chinese Academy of Sciences, and Biomedical Core Facility of Fudan University for the support on cryo-EM data collection and data analyses. This work was supported by the Ministry of Science and Technology of China (2016YFA0500700 and 2016YFA0501100), the National Natural Science Foundation of China (U1432242, 31425008 and 91419301), the National Program for support of Top-Notch Young Professionals (YX), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB08000000), and the Beijing Municipal Science & Technology Commission (Z161100000116034 to HW).
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( Supplementary information is linked to the online version of the paper on the Cell Research website.)
Supplementary information
Supplementary information, Figure S1
Complex purification and characterization of ATR-ATRIP. (PDF 607 kb)
Supplementary information, Figure S2
Cryo-EM structure determination of ATR-ATRIP. (PDF 464 kb)
Supplementary information, Figure S3
Cryo-EM map for the ATR-ATRIP complex. (PDF 1942 kb)
Supplementary information, Figure S4
Sequence alignment of ATR across species. (PDF 1940 kb)
Supplementary information, Figure S5
Structural comparison of ATR, mTOR and ATM. (PDF 657 kb)
Supplementary information, Figure S6
Two ATR monomers for the ATR-ATRIP complex formation. (PDF 919 kb)
Supplementary information, Figure S7
Interactions between ATR and ATRIP in the complex. (PDF 454 kb)
Supplementary information, Movie S1
EM map and model building of ATR-ATRIP complex. (MP4 14051 kb)
Supplementary information, Movie S2
Structural analysis of ATR-ATRIP complex. (MP4 73867 kb)
Supplementary information, Table S1
Statistics for data collection and model refinement of ATR-ATRIP complex. (PDF 60 kb)
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Rao, Q., Liu, M., Tian, Y. et al. Cryo-EM structure of human ATR-ATRIP complex. Cell Res 28, 143–156 (2018). https://doi.org/10.1038/cr.2017.158
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DOI: https://doi.org/10.1038/cr.2017.158
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