Biochemical and structural characterization of analogs of MRE11 breast cancer-associated mutant F237C

Abstract

The MRE11–RAD50–NBS1 (MRN) protein complex plays a vital role in DNA double strand break sensing, signaling, and repair. Mutation in any component of this complex may lead to disease as disrupting DNA double strand break repair has the potential to cause translocations and loss of genomic information. Here, we have investigated an MRE11 mutation, F237C, identified in a breast cancer tumor. We found that the analogous mutant of Pyrococcus furiosus Mre11 diminishes both the exonuclease and endonuclease activities of Mre11 in vitro. Solution state NMR experiments show that this mutant causes structural changes in the DNA-bound Mre11 for both exo- and endonuclease substrates and causes the protein to become generally more rigid. Moreover, by comparing the NMR data for this cancer-associated mutant with two previously described Mre11 separation-of-nuclease function mutants, a potential allosteric network was detected within Mre11 that connects the active site to regions responsible for recognizing the DNA ends and for dimerization. Together, our data further highlight the dynamics required for Mre11 nuclease function and illuminate the presence of allostery within the enzyme.

Description

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Keywords

DNA Repair Enzymes, NMR Spectroscopy

Citation

Rahman, S., Beikzadeh, M. & Latham, M.P. Biochemical and structural characterization of analogs of MRE11 breast cancer-associated mutant F237C. Sci Rep 11, 7089 (2021). https://doi.org/10.1038/s41598-021-86552-0

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