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Backbone and ILV side-chain methyl NMR resonance assignments of human Rev7/Rev3-RBM1 and Rev7/Rev3-RBM2 complexes

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Abstract

Rev7 is a versatile HORMA (Hop1, Rev7, Mad2) family adaptor protein with multiple roles in mitotic regulation and DNA damage response, and an essential accessory subunit of the translesion synthesis (TLS) DNA polymerase Polζ employed in replication of damaged DNA. Within Polζ, the two copies of Rev7 interact with the two Rev7-bonding motifs (RBM1 and RBM2) of the catalytic subunit Rev3 by a mechanism characteristic of HORMA proteins whereby the “safety-belt” loop of Rev7 closes on the top of the ligand. Here we report the nearly complete backbone and Ile, Val, Leu side-chain methyl NMR resonance assignments of the 27 kDa human Rev7/Rev3-RBM1 and Rev7/Rev3-RBM2 complexes (BMRB deposition numbers 51651 and 51652) that will facilitate future NMR studies of Rev7 dynamics and interactions.

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Acknowledgements

This work was supported by the National Institutes of Health R01CA233959 grant to D.M.K. and M. Kyle Hadden. The authors thank Dr. Lewis E. Kay for sharing library of custom pulse-sequences used in this work, Dr. Irina Semenova for assistance in generating the Rev7 triple-mutant constructs, and Mr. Sam Mahdi for discussion.

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  1. Department of Molecular Biology and Biophysics, University of Connecticut Health Center, Farmington, CT, 06030, USA

    Gianluca A. Arianna, Dane H. Geddes-Buehre & Dmitry M. Korzhnev

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  1. Gianluca A. Arianna
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  2. Dane H. Geddes-Buehre
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  3. Dmitry M. Korzhnev
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Contributions

D.H.G. and G.A.A. produced the proteins and performed assignments for the Rev7/Rev3-RBM1 and Rev7/Rev3-RBM2 complexes, respectively. D.M.K. performed NMR experiments. All authors contributed to the manuscript preparation.

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Correspondence to Dmitry M. Korzhnev.

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Arianna, G.A., Geddes-Buehre, D.H. & Korzhnev, D.M. Backbone and ILV side-chain methyl NMR resonance assignments of human Rev7/Rev3-RBM1 and Rev7/Rev3-RBM2 complexes. Biomol NMR Assign 17, 107–114 (2023). https://doi.org/10.1007/s12104-023-10128-4

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