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Analysis of diverse double-strand break synapsis with Polλ reveals basis for unique substrate specificity in nonhomologous end-joining

DNA double-strand breaks (DSBs) threaten genomic stability, since their persistence can lead to loss of critical genetic information, chromosomal translocations or rearrangements, and cell death. DSBs can be repaired through the nonhomologous end-joining pathway (NHEJ), which processes and ligates D...

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Autores principales: Kaminski, Andrea M., Chiruvella, Kishore K., Ramsden, Dale A., Bebenek, Katarzyna, Kunkel, Thomas A., Pedersen, Lars C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9249759/
https://www.ncbi.nlm.nih.gov/pubmed/35778389
http://dx.doi.org/10.1038/s41467-022-31278-4
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author Kaminski, Andrea M.
Chiruvella, Kishore K.
Ramsden, Dale A.
Bebenek, Katarzyna
Kunkel, Thomas A.
Pedersen, Lars C.
author_facet Kaminski, Andrea M.
Chiruvella, Kishore K.
Ramsden, Dale A.
Bebenek, Katarzyna
Kunkel, Thomas A.
Pedersen, Lars C.
author_sort Kaminski, Andrea M.
collection PubMed
description DNA double-strand breaks (DSBs) threaten genomic stability, since their persistence can lead to loss of critical genetic information, chromosomal translocations or rearrangements, and cell death. DSBs can be repaired through the nonhomologous end-joining pathway (NHEJ), which processes and ligates DNA ends efficiently to prevent or minimize sequence loss. Polymerase λ (Polλ), one of the Family X polymerases, fills sequence gaps of DSB substrates with a strict specificity for a base-paired primer terminus. There is little information regarding Polλ’s approach to engaging such substrates. We used in vitro polymerization and cell-based NHEJ assays to explore the contributions of conserved loop regions toward DSB substrate specificity and utilization. In addition, we present multiple crystal structures of Polλ in synapsis with varying biologically relevant DSB end configurations, revealing how key structural features and hydrogen bonding networks work in concert to stabilize these tenuous, potentially cytotoxic DNA lesions during NHEJ.
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spelling pubmed-92497592022-07-03 Analysis of diverse double-strand break synapsis with Polλ reveals basis for unique substrate specificity in nonhomologous end-joining Kaminski, Andrea M. Chiruvella, Kishore K. Ramsden, Dale A. Bebenek, Katarzyna Kunkel, Thomas A. Pedersen, Lars C. Nat Commun Article DNA double-strand breaks (DSBs) threaten genomic stability, since their persistence can lead to loss of critical genetic information, chromosomal translocations or rearrangements, and cell death. DSBs can be repaired through the nonhomologous end-joining pathway (NHEJ), which processes and ligates DNA ends efficiently to prevent or minimize sequence loss. Polymerase λ (Polλ), one of the Family X polymerases, fills sequence gaps of DSB substrates with a strict specificity for a base-paired primer terminus. There is little information regarding Polλ’s approach to engaging such substrates. We used in vitro polymerization and cell-based NHEJ assays to explore the contributions of conserved loop regions toward DSB substrate specificity and utilization. In addition, we present multiple crystal structures of Polλ in synapsis with varying biologically relevant DSB end configurations, revealing how key structural features and hydrogen bonding networks work in concert to stabilize these tenuous, potentially cytotoxic DNA lesions during NHEJ. Nature Publishing Group UK 2022-07-01 /pmc/articles/PMC9249759/ /pubmed/35778389 http://dx.doi.org/10.1038/s41467-022-31278-4 Text en © This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2022 https://creativecommons.org/licenses/by/4.0/Open Access This 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Kaminski, Andrea M.
Chiruvella, Kishore K.
Ramsden, Dale A.
Bebenek, Katarzyna
Kunkel, Thomas A.
Pedersen, Lars C.
Analysis of diverse double-strand break synapsis with Polλ reveals basis for unique substrate specificity in nonhomologous end-joining
title Analysis of diverse double-strand break synapsis with Polλ reveals basis for unique substrate specificity in nonhomologous end-joining
title_full Analysis of diverse double-strand break synapsis with Polλ reveals basis for unique substrate specificity in nonhomologous end-joining
title_fullStr Analysis of diverse double-strand break synapsis with Polλ reveals basis for unique substrate specificity in nonhomologous end-joining
title_full_unstemmed Analysis of diverse double-strand break synapsis with Polλ reveals basis for unique substrate specificity in nonhomologous end-joining
title_short Analysis of diverse double-strand break synapsis with Polλ reveals basis for unique substrate specificity in nonhomologous end-joining
title_sort analysis of diverse double-strand break synapsis with polλ reveals basis for unique substrate specificity in nonhomologous end-joining
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9249759/
https://www.ncbi.nlm.nih.gov/pubmed/35778389
http://dx.doi.org/10.1038/s41467-022-31278-4
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