Journal article
Structural basis for multifunctional roles of human Ints3 C-terminal domain
The Journal of biological chemistry, Vol.296(C), pp.100112-100112
01/01/2021
DOI: 10.1074/jbc.RA120.016393
PMCID: PMC7948952
PMID: 33434574
Abstract
Proper repair of damaged DNA is critical for the maintenance of genome stability. A complex composed of Integrator subunit 3 (Ints3), single-stranded DNA-binding protein 1 (SSB1), and SSB-interacting protein 1 (SSBIP1) is required for efficient homologous recombination-dependent repair of double-strand breaks (DSBs) and ataxia-telangiectasia mutated (ATM)-dependent signaling pathways. It is known that in this complex the Ints3 N-terminal domain scaffolds SSB1 and SSBIP1. However, the molecular basis for the function of the Ints3 C-terminal domain remains unclear. Here, we present the crystal structure of the Ints3 C-terminal domain, uncovering a HEAT-repeat superhelical fold. Using structure and mutation analysis, we show that the C-terminal domain exists as a stable dimer. A basic groove and a cluster of conserved residues on two opposite sides of the dimer bind single-stranded RNA/DNA (ssRNA/ssDNA) and Integrator complex subunit 6 (Ints6), respectively. Dimerization is required for nucleic acid binding, but not for Ints6 binding. Additionally, in vitro experiments using HEK 293T cells demonstrate that Ints6 interaction is critical for maintaining SSB1 protein level. Taken together, our findings establish the structural basis of a multifunctional Ints3 C-terminal module, allowing us to propose a novel mode of nucleic acid recognition by helical repeat protein and paving the way for future mechanistic studies.
Details
- Title: Subtitle
- Structural basis for multifunctional roles of human Ints3 C-terminal domain
- Creators
- Jian Li - China-US (Henan) Hormel Cancer InstituteXinli Ma - University of MinnesotaSurajit Banerjee - Cornell UniversitySankar Baruah - University of IowaNicholas J. Schnicker - University of IowaEunmiri Roh - Kwangju Women's UniversityWeiya Ma - HormelKangdong Liu - China-US (Henan) Hormel Cancer InstituteAnn M. Bode - HormelZigang Dong - China-US (Henan) Hormel Cancer Institute
- Resource Type
- Journal article
- Publication Details
- The Journal of biological chemistry, Vol.296(C), pp.100112-100112
- Publisher
- Elsevier Inc
- DOI
- 10.1074/jbc.RA120.016393
- PMID
- 33434574
- PMCID
- PMC7948952
- ISSN
- 0021-9258
- eISSN
- 1083-351X
- Language
- English
- Date published
- 01/01/2021
- Academic Unit
- Molecular Physiology and Biophysics; Medicine Administration
- Record Identifier
- 9984622753302771
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