The DUF328 Family Member Yaaa is a Dna-Binding Protein with a Novel Fold

Authors

Janani Prahlad, University of Nebraska - Lincoln
Yifeng Yuan, University of Florida
Jiusheng Lin, University of Nebraska - Lincoln
Chou-Wei Chang, Beckman Research Institute of City of Hope
Dirk Iwata-Reuyl, Portland State UniversityFollow
Yilun Liu, Beckman Research Institute of City of Hope
Valérie de Crécy-Lagard, University of Florida
Mark A. Wilson, University of Nebraska - Lincoln

Published In

The Journal of Biological Chemistry

Document Type

Citation

Publication Date

8-12-2020

Abstract

DUF328 family proteins are present in many prokaryotes, however their molecular activities are unknown. The Escherichia coli DUF328 protein YaaA is a member of the OxyR regulon and is protective against oxidative stress. Because uncharacterized proteins involved in prokaryotic oxidative stress response are rare, we sought to learn more about the DUF328 family. Using comparative genomics, we found a robust association between the DUF328 family and genes involved in DNA recombination and the oxidative stress response. In some proteins, DUF328 domains are fused to other domains involved in DNA binding, recombination, and repair. Co-fitness analysis indicates that DUF328 family genes associate with recombination-mediated DNA repair pathways, particularly the RecFOR pathway. Purified recombinant YaaA binds to double-stranded DNA, duplex DNA containing bubbles of unpaired nucleotides, and Holliday junction constructs in vitro with dissociation equilibrium constants of 200-300 nM. YaaA binds DNA with positive cooperativity, forming multiple shifted species in electrophoretic mobility shift assays. The 1.65 Å resolution X-ray crystal structure of YaaA reveals that the protein possesses a new fold that we name the cantaloupe fold. YaaA has a positively charged cleft and a helix-hairpin-helix (HhH) DNA binding motif found in other DNA repair enzymes. Our results demonstrate that YaaA is a new type of DNA-binding protein associated with the oxidative stress response and that this molecular function is likely conserved in other DUF328 family members.

Description

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Locate the Document

https://doi.org/10.1074/jbc.RA120.015055

DOI

10.1074/jbc.RA120.015055

Persistent Identifier

https://archives.pdx.edu/ds/psu/33972

Citation Details

Prahlad, J., Yuan, Y., Lin, J., Chang, C.-W., Iwata-Reuyl, D., Liu, Y., de Crécy-Lagard, V., & Wilson, M. A. (2020). The DUF328 family member YaaA is a DNA-binding protein with a novel fold. Journal of Biological Chemistry, jbc.RA120.015055. https://doi.org/10.1074/JBC.RA120.015055