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Prolyl Isomerization-Mediated Conformational Changes Define ATR Subcellular Compartment-Specific Functions
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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9204103/ https://www.ncbi.nlm.nih.gov/pubmed/35721505 http://dx.doi.org/10.3389/fcell.2022.826576 |
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author | Biswas, Himadri Zhao, Shu-Jun Makinwa, Yetunde Bassett, James S. Musich, Phillip R. Liu, Jing-Yuan Zou, Yue |
author_facet | Biswas, Himadri Zhao, Shu-Jun Makinwa, Yetunde Bassett, James S. Musich, Phillip R. Liu, Jing-Yuan Zou, Yue |
author_sort | Biswas, Himadri |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-9204103 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-92041032022-06-18 Prolyl Isomerization-Mediated Conformational Changes Define ATR Subcellular Compartment-Specific Functions Biswas, Himadri Zhao, Shu-Jun Makinwa, Yetunde Bassett, James S. Musich, Phillip R. Liu, Jing-Yuan Zou, Yue Front Cell Dev Biol Cell and Developmental Biology 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. Frontiers Media S.A. 2022-06-03 /pmc/articles/PMC9204103/ /pubmed/35721505 http://dx.doi.org/10.3389/fcell.2022.826576 Text en Copyright © 2022 Biswas, Zhao, Makinwa, Bassett, Musich, Liu and Zou. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Cell and Developmental Biology Biswas, Himadri Zhao, Shu-Jun Makinwa, Yetunde Bassett, James S. Musich, Phillip R. Liu, Jing-Yuan Zou, Yue Prolyl Isomerization-Mediated Conformational Changes Define ATR Subcellular Compartment-Specific Functions |
title | Prolyl Isomerization-Mediated Conformational Changes Define ATR Subcellular Compartment-Specific Functions |
title_full | Prolyl Isomerization-Mediated Conformational Changes Define ATR Subcellular Compartment-Specific Functions |
title_fullStr | Prolyl Isomerization-Mediated Conformational Changes Define ATR Subcellular Compartment-Specific Functions |
title_full_unstemmed | Prolyl Isomerization-Mediated Conformational Changes Define ATR Subcellular Compartment-Specific Functions |
title_short | Prolyl Isomerization-Mediated Conformational Changes Define ATR Subcellular Compartment-Specific Functions |
title_sort | prolyl isomerization-mediated conformational changes define atr subcellular compartment-specific functions |
topic | Cell and Developmental Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9204103/ https://www.ncbi.nlm.nih.gov/pubmed/35721505 http://dx.doi.org/10.3389/fcell.2022.826576 |
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