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(1)H, (13)C, (15)N backbone resonance assignment for the 1–164 construct of human XRCC4
DNA double-strand breaks (DSBs) represent the most cytotoxic DNA lesions, as—if mis- or unrepaired—they can cause cell death or lead to genome instability, which in turn can cause cancer. DSBs are repaired by two major pathways termed homologous recombination and non-homologous end-joining (NHEJ). N...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Springer Netherlands
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8481219/ https://www.ncbi.nlm.nih.gov/pubmed/34173222 http://dx.doi.org/10.1007/s12104-021-10035-6 |
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author | Cabello-Lobato, Maria Jose Schmidt, Christine K. Cliff, Matthew J. |
author_facet | Cabello-Lobato, Maria Jose Schmidt, Christine K. Cliff, Matthew J. |
author_sort | Cabello-Lobato, Maria Jose |
collection | PubMed |
description | DNA double-strand breaks (DSBs) represent the most cytotoxic DNA lesions, as—if mis- or unrepaired—they can cause cell death or lead to genome instability, which in turn can cause cancer. DSBs are repaired by two major pathways termed homologous recombination and non-homologous end-joining (NHEJ). NHEJ is responsible for repairing the vast majority of DSBs arising in human cells. Defects in NHEJ factors are also associated with microcephaly, primordial dwarfism and immune deficiencies. One of the key proteins important for mediating NHEJ is XRCC4. XRCC4 is a dimer, with the dimer interface mediated by an extended coiled-coil. The N-terminal head domain forms a mixed alpha–beta globular structure. Numerous factors interact with the C-terminus of the coiled-coil domain, which is also associated with significant self-association between XRCC4 dimers. A range of construct lengths of human XRCC4 were expressed and purified, and the 1–164 variant had the best NMR properties, as judged by consistent linewidths, and chemical shift dispersion. In this work we report the (1)H, (15) N and (13)C backbone resonance assignments of human XRCC4 in the solution form of the 1–164 construct. Assignments were obtained by heteronuclear multidimensional NMR spectroscopy. In total, 156 of 161 assignable residues of XRCC4 were assigned to resonances in the TROSY spectrum, with an additional 11 resonances assigned to His-Tag residues. Prediction of solution secondary structure from a chemical shift analysis using the TALOS + webserver is in good agreement with the published X-ray crystal structures of this protein. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12104-021-10035-6. |
format | Online Article Text |
id | pubmed-8481219 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Springer Netherlands |
record_format | MEDLINE/PubMed |
spelling | pubmed-84812192021-10-08 (1)H, (13)C, (15)N backbone resonance assignment for the 1–164 construct of human XRCC4 Cabello-Lobato, Maria Jose Schmidt, Christine K. Cliff, Matthew J. Biomol NMR Assign Article DNA double-strand breaks (DSBs) represent the most cytotoxic DNA lesions, as—if mis- or unrepaired—they can cause cell death or lead to genome instability, which in turn can cause cancer. DSBs are repaired by two major pathways termed homologous recombination and non-homologous end-joining (NHEJ). NHEJ is responsible for repairing the vast majority of DSBs arising in human cells. Defects in NHEJ factors are also associated with microcephaly, primordial dwarfism and immune deficiencies. One of the key proteins important for mediating NHEJ is XRCC4. XRCC4 is a dimer, with the dimer interface mediated by an extended coiled-coil. The N-terminal head domain forms a mixed alpha–beta globular structure. Numerous factors interact with the C-terminus of the coiled-coil domain, which is also associated with significant self-association between XRCC4 dimers. A range of construct lengths of human XRCC4 were expressed and purified, and the 1–164 variant had the best NMR properties, as judged by consistent linewidths, and chemical shift dispersion. In this work we report the (1)H, (15) N and (13)C backbone resonance assignments of human XRCC4 in the solution form of the 1–164 construct. Assignments were obtained by heteronuclear multidimensional NMR spectroscopy. In total, 156 of 161 assignable residues of XRCC4 were assigned to resonances in the TROSY spectrum, with an additional 11 resonances assigned to His-Tag residues. Prediction of solution secondary structure from a chemical shift analysis using the TALOS + webserver is in good agreement with the published X-ray crystal structures of this protein. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12104-021-10035-6. Springer Netherlands 2021-06-25 2021 /pmc/articles/PMC8481219/ /pubmed/34173222 http://dx.doi.org/10.1007/s12104-021-10035-6 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Cabello-Lobato, Maria Jose Schmidt, Christine K. Cliff, Matthew J. (1)H, (13)C, (15)N backbone resonance assignment for the 1–164 construct of human XRCC4 |
title | (1)H, (13)C, (15)N backbone resonance assignment for the 1–164 construct of human XRCC4 |
title_full | (1)H, (13)C, (15)N backbone resonance assignment for the 1–164 construct of human XRCC4 |
title_fullStr | (1)H, (13)C, (15)N backbone resonance assignment for the 1–164 construct of human XRCC4 |
title_full_unstemmed | (1)H, (13)C, (15)N backbone resonance assignment for the 1–164 construct of human XRCC4 |
title_short | (1)H, (13)C, (15)N backbone resonance assignment for the 1–164 construct of human XRCC4 |
title_sort | (1)h, (13)c, (15)n backbone resonance assignment for the 1–164 construct of human xrcc4 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8481219/ https://www.ncbi.nlm.nih.gov/pubmed/34173222 http://dx.doi.org/10.1007/s12104-021-10035-6 |
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