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Evolution of Rev7 interactions in eukaryotic TLS DNA polymerase Polζ

Translesion synthesis (TLS) DNA polymerase Polζ is crucial for the bypass replication over sites of DNA damage. The Rev7 subunit of Polζ is a HORMA (Hop1, Rev7, Mad2) protein that facilitates recruitment of Polζ to the replication fork via interactions with the catalytic subunit Rev3 and the transle...

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Autores principales: McPherson, Kerry Silva, Rizzo, Alessandro A., Erlandsen, Heidi, Chatterjee, Nimrat, Walker, Graham C., Korzhnev, Dmitry M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9926120/
https://www.ncbi.nlm.nih.gov/pubmed/36592930
http://dx.doi.org/10.1016/j.jbc.2022.102859
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author McPherson, Kerry Silva
Rizzo, Alessandro A.
Erlandsen, Heidi
Chatterjee, Nimrat
Walker, Graham C.
Korzhnev, Dmitry M.
author_facet McPherson, Kerry Silva
Rizzo, Alessandro A.
Erlandsen, Heidi
Chatterjee, Nimrat
Walker, Graham C.
Korzhnev, Dmitry M.
author_sort McPherson, Kerry Silva
collection PubMed
description Translesion synthesis (TLS) DNA polymerase Polζ is crucial for the bypass replication over sites of DNA damage. The Rev7 subunit of Polζ is a HORMA (Hop1, Rev7, Mad2) protein that facilitates recruitment of Polζ to the replication fork via interactions with the catalytic subunit Rev3 and the translesion synthesis scaffold protein Rev1. Human Rev7 (hRev7) interacts with two Rev7-binding motifs (RBMs) of hRev3 by a mechanism conserved among HORMA proteins whereby the safety-belt loop of hRev7 closes on the top of the ligand. The two copies of hRev7 tethered by the two hRev3-RBMs form a symmetric head-to-head dimer through the canonical HORMA dimerization interface. Recent cryo-EM structures reveal that Saccharomyces cerevisiae Polζ (scPolζ) also includes two copies of scRev7 bound to distinct regions of scRev3. Surprisingly, the HORMA dimerization interface is not conserved in scRev7, with the two scRev7 protomers forming an asymmetric head-to-tail dimer with a much smaller interface than the hRev7 dimer. Here, we validated the two adjacent RBM motifs in scRev3, which bind scRev7 with affinities that differ by two orders of magnitude and confirmed the 2:1 stoichiometry of the scRev7:Rev3 complex in solution. However, our biophysical studies reveal that scRev7 does not form dimers in solution either on its own accord or when tethered by the two RBMs in scRev3. These findings imply that the scRev7 dimer observed in the cryo-EM structures is induced by scRev7 interactions with other Polζ subunits and that Rev7 homodimerization via the HORMA interface is a mechanism that emerged later in evolution.
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spelling pubmed-99261202023-02-16 Evolution of Rev7 interactions in eukaryotic TLS DNA polymerase Polζ McPherson, Kerry Silva Rizzo, Alessandro A. Erlandsen, Heidi Chatterjee, Nimrat Walker, Graham C. Korzhnev, Dmitry M. J Biol Chem Research Article Translesion synthesis (TLS) DNA polymerase Polζ is crucial for the bypass replication over sites of DNA damage. The Rev7 subunit of Polζ is a HORMA (Hop1, Rev7, Mad2) protein that facilitates recruitment of Polζ to the replication fork via interactions with the catalytic subunit Rev3 and the translesion synthesis scaffold protein Rev1. Human Rev7 (hRev7) interacts with two Rev7-binding motifs (RBMs) of hRev3 by a mechanism conserved among HORMA proteins whereby the safety-belt loop of hRev7 closes on the top of the ligand. The two copies of hRev7 tethered by the two hRev3-RBMs form a symmetric head-to-head dimer through the canonical HORMA dimerization interface. Recent cryo-EM structures reveal that Saccharomyces cerevisiae Polζ (scPolζ) also includes two copies of scRev7 bound to distinct regions of scRev3. Surprisingly, the HORMA dimerization interface is not conserved in scRev7, with the two scRev7 protomers forming an asymmetric head-to-tail dimer with a much smaller interface than the hRev7 dimer. Here, we validated the two adjacent RBM motifs in scRev3, which bind scRev7 with affinities that differ by two orders of magnitude and confirmed the 2:1 stoichiometry of the scRev7:Rev3 complex in solution. However, our biophysical studies reveal that scRev7 does not form dimers in solution either on its own accord or when tethered by the two RBMs in scRev3. These findings imply that the scRev7 dimer observed in the cryo-EM structures is induced by scRev7 interactions with other Polζ subunits and that Rev7 homodimerization via the HORMA interface is a mechanism that emerged later in evolution. American Society for Biochemistry and Molecular Biology 2022-12-31 /pmc/articles/PMC9926120/ /pubmed/36592930 http://dx.doi.org/10.1016/j.jbc.2022.102859 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research Article
McPherson, Kerry Silva
Rizzo, Alessandro A.
Erlandsen, Heidi
Chatterjee, Nimrat
Walker, Graham C.
Korzhnev, Dmitry M.
Evolution of Rev7 interactions in eukaryotic TLS DNA polymerase Polζ
title Evolution of Rev7 interactions in eukaryotic TLS DNA polymerase Polζ
title_full Evolution of Rev7 interactions in eukaryotic TLS DNA polymerase Polζ
title_fullStr Evolution of Rev7 interactions in eukaryotic TLS DNA polymerase Polζ
title_full_unstemmed Evolution of Rev7 interactions in eukaryotic TLS DNA polymerase Polζ
title_short Evolution of Rev7 interactions in eukaryotic TLS DNA polymerase Polζ
title_sort evolution of rev7 interactions in eukaryotic tls dna polymerase polζ
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9926120/
https://www.ncbi.nlm.nih.gov/pubmed/36592930
http://dx.doi.org/10.1016/j.jbc.2022.102859
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