Cargando…
Ataxia Telangiectasia-Mutated (ATM) Kinase Activity Is Regulated by ATP-driven Conformational Changes in the Mre11/Rad50/Nbs1 (MRN) Complex
The Ataxia Telangiectasia-Mutated (ATM) protein kinase is recruited to sites of double-strand DNA breaks by the Mre11/Rad50/Nbs1 (MRN) complex, which also facilitates ATM monomerization and activation. MRN exists in at least two distinct conformational states, dependent on ATP binding and hydrolysis...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2013
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3642328/ https://www.ncbi.nlm.nih.gov/pubmed/23525106 http://dx.doi.org/10.1074/jbc.M113.460378 |
_version_ | 1782268128698302464 |
---|---|
author | Lee, Ji-Hoon Mand, Michael R. Deshpande, Rajashree A. Kinoshita, Eri Yang, Soo-Hyun Wyman, Claire Paull, Tanya T. |
author_facet | Lee, Ji-Hoon Mand, Michael R. Deshpande, Rajashree A. Kinoshita, Eri Yang, Soo-Hyun Wyman, Claire Paull, Tanya T. |
author_sort | Lee, Ji-Hoon |
collection | PubMed |
description | The Ataxia Telangiectasia-Mutated (ATM) protein kinase is recruited to sites of double-strand DNA breaks by the Mre11/Rad50/Nbs1 (MRN) complex, which also facilitates ATM monomerization and activation. MRN exists in at least two distinct conformational states, dependent on ATP binding and hydrolysis by the Rad50 protein. Here we use an ATP analog-sensitive form of ATM to determine that ATP binding, but not hydrolysis, by Rad50 is essential for MRN stimulation of ATM. Mre11 nuclease activity is dispensable, although some mutations in the Mre11 catalytic domain block ATM activation independent of nuclease function, as does the mirin compound. The coiled-coil domains of Rad50 are important for the DNA binding ability of MRN and are essential for ATM activation, but loss of the zinc hook connection can be substituted by higher levels of the complex. Nbs1 binds to the “closed” form of the MR complex, promoted by the zinc hook and by ATP binding. Thus the primary role of the hook is to tether Rad50 monomers together, promoting the association of the Rad50 catalytic domains into a form that binds ATP and also binds Nbs1. Collectively, these results show that the ATP-bound form of MRN is the critical conformation for ATM activation. |
format | Online Article Text |
id | pubmed-3642328 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-36423282013-05-08 Ataxia Telangiectasia-Mutated (ATM) Kinase Activity Is Regulated by ATP-driven Conformational Changes in the Mre11/Rad50/Nbs1 (MRN) Complex Lee, Ji-Hoon Mand, Michael R. Deshpande, Rajashree A. Kinoshita, Eri Yang, Soo-Hyun Wyman, Claire Paull, Tanya T. J Biol Chem DNA and Chromosomes The Ataxia Telangiectasia-Mutated (ATM) protein kinase is recruited to sites of double-strand DNA breaks by the Mre11/Rad50/Nbs1 (MRN) complex, which also facilitates ATM monomerization and activation. MRN exists in at least two distinct conformational states, dependent on ATP binding and hydrolysis by the Rad50 protein. Here we use an ATP analog-sensitive form of ATM to determine that ATP binding, but not hydrolysis, by Rad50 is essential for MRN stimulation of ATM. Mre11 nuclease activity is dispensable, although some mutations in the Mre11 catalytic domain block ATM activation independent of nuclease function, as does the mirin compound. The coiled-coil domains of Rad50 are important for the DNA binding ability of MRN and are essential for ATM activation, but loss of the zinc hook connection can be substituted by higher levels of the complex. Nbs1 binds to the “closed” form of the MR complex, promoted by the zinc hook and by ATP binding. Thus the primary role of the hook is to tether Rad50 monomers together, promoting the association of the Rad50 catalytic domains into a form that binds ATP and also binds Nbs1. Collectively, these results show that the ATP-bound form of MRN is the critical conformation for ATM activation. American Society for Biochemistry and Molecular Biology 2013-05-03 2013-03-22 /pmc/articles/PMC3642328/ /pubmed/23525106 http://dx.doi.org/10.1074/jbc.M113.460378 Text en © 2013 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version full access. Creative Commons Attribution Unported License (http://creativecommons.org/licenses/by/3.0/) applies to Author Choice Articles |
spellingShingle | DNA and Chromosomes Lee, Ji-Hoon Mand, Michael R. Deshpande, Rajashree A. Kinoshita, Eri Yang, Soo-Hyun Wyman, Claire Paull, Tanya T. Ataxia Telangiectasia-Mutated (ATM) Kinase Activity Is Regulated by ATP-driven Conformational Changes in the Mre11/Rad50/Nbs1 (MRN) Complex |
title | Ataxia Telangiectasia-Mutated (ATM) Kinase Activity Is Regulated by ATP-driven Conformational Changes in the Mre11/Rad50/Nbs1 (MRN) Complex |
title_full | Ataxia Telangiectasia-Mutated (ATM) Kinase Activity Is Regulated by ATP-driven Conformational Changes in the Mre11/Rad50/Nbs1 (MRN) Complex |
title_fullStr | Ataxia Telangiectasia-Mutated (ATM) Kinase Activity Is Regulated by ATP-driven Conformational Changes in the Mre11/Rad50/Nbs1 (MRN) Complex |
title_full_unstemmed | Ataxia Telangiectasia-Mutated (ATM) Kinase Activity Is Regulated by ATP-driven Conformational Changes in the Mre11/Rad50/Nbs1 (MRN) Complex |
title_short | Ataxia Telangiectasia-Mutated (ATM) Kinase Activity Is Regulated by ATP-driven Conformational Changes in the Mre11/Rad50/Nbs1 (MRN) Complex |
title_sort | ataxia telangiectasia-mutated (atm) kinase activity is regulated by atp-driven conformational changes in the mre11/rad50/nbs1 (mrn) complex |
topic | DNA and Chromosomes |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3642328/ https://www.ncbi.nlm.nih.gov/pubmed/23525106 http://dx.doi.org/10.1074/jbc.M113.460378 |
work_keys_str_mv | AT leejihoon ataxiatelangiectasiamutatedatmkinaseactivityisregulatedbyatpdrivenconformationalchangesinthemre11rad50nbs1mrncomplex AT mandmichaelr ataxiatelangiectasiamutatedatmkinaseactivityisregulatedbyatpdrivenconformationalchangesinthemre11rad50nbs1mrncomplex AT deshpanderajashreea ataxiatelangiectasiamutatedatmkinaseactivityisregulatedbyatpdrivenconformationalchangesinthemre11rad50nbs1mrncomplex AT kinoshitaeri ataxiatelangiectasiamutatedatmkinaseactivityisregulatedbyatpdrivenconformationalchangesinthemre11rad50nbs1mrncomplex AT yangsoohyun ataxiatelangiectasiamutatedatmkinaseactivityisregulatedbyatpdrivenconformationalchangesinthemre11rad50nbs1mrncomplex AT wymanclaire ataxiatelangiectasiamutatedatmkinaseactivityisregulatedbyatpdrivenconformationalchangesinthemre11rad50nbs1mrncomplex AT paulltanyat ataxiatelangiectasiamutatedatmkinaseactivityisregulatedbyatpdrivenconformationalchangesinthemre11rad50nbs1mrncomplex |