Structure of the human DNA-repair protein RAD52 containing surface mutations

Mika Saotome; Kengo Saito; Keiichi Onodera; Hitoshi Kurumizaka; Wataru Kagawa

doi:10.1107/S2053230X1601027X

Structure of the human DNA-repair protein RAD52 containing surface mutations

Mika Saotome, Kengo Saito, Keiichi Onodera, Hitoshi Kurumizaka, Wataru Kagawa^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

2 Citations (Scopus)

Abstract

The Rad52 protein is a eukaryotic single-strand DNA-annealing protein that is involved in the homologous recombinational repair of DNA double-strand breaks. The isolated N-terminal half of the human RAD52 protein (RAD52^1-212) forms an undecameric ring structure with a surface that is mostly positively charged. In the present study, it was found that RAD52^1-212 containing alanine mutations of the charged surface residues (Lys102, Lys133 and Glu202) is highly amenable to crystallization. The structure of the mutant RAD52^1-212 was solved at 2.4 Å resolution. The structure revealed an association between the symmetry-related RAD52^1-212 rings, in which a partially unfolded, C-terminal region of RAD52 extended into the DNA-binding groove of the neighbouring ring in the crystal. The alanine mutations probably reduced the surface entropy of the RAD52^1-212 ring and stabilized the ring-ring association observed in the crystal.

Original language	English
Pages (from-to)	598-603
Number of pages	6
Journal	Acta Crystallographica Section F:Structural Biology Communications
Volume	72
DOIs	https://doi.org/10.1107/S2053230X1601027X
Publication status	Published - 2016

Keywords

higher order interaction
homologous recombinational repair
RAD52
single-strand annealing proteins
ssDNA-binding protein
surface-entropy reduction

ASJC Scopus subject areas

Structural Biology
Biophysics
Biochemistry
Genetics
Condensed Matter Physics

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10.1107/S2053230X1601027X

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title = "Structure of the human DNA-repair protein RAD52 containing surface mutations",

abstract = "The Rad52 protein is a eukaryotic single-strand DNA-annealing protein that is involved in the homologous recombinational repair of DNA double-strand breaks. The isolated N-terminal half of the human RAD52 protein (RAD521-212) forms an undecameric ring structure with a surface that is mostly positively charged. In the present study, it was found that RAD521-212 containing alanine mutations of the charged surface residues (Lys102, Lys133 and Glu202) is highly amenable to crystallization. The structure of the mutant RAD521-212 was solved at 2.4 {\AA} resolution. The structure revealed an association between the symmetry-related RAD521-212 rings, in which a partially unfolded, C-terminal region of RAD52 extended into the DNA-binding groove of the neighbouring ring in the crystal. The alanine mutations probably reduced the surface entropy of the RAD521-212 ring and stabilized the ring-ring association observed in the crystal.",

keywords = "higher order interaction, homologous recombinational repair, RAD52, single-strand annealing proteins, ssDNA-binding protein, surface-entropy reduction",

author = "Mika Saotome and Kengo Saito and Keiichi Onodera and Hitoshi Kurumizaka and Wataru Kagawa",

year = "2016",

doi = "10.1107/S2053230X1601027X",

language = "English",

volume = "72",

pages = "598--603",

journal = "Acta Crystallographica Section F:Structural Biology Communications",

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publisher = "John Wiley and Sons Ltd",

}

TY - JOUR

T1 - Structure of the human DNA-repair protein RAD52 containing surface mutations

AU - Saotome, Mika

AU - Saito, Kengo

AU - Onodera, Keiichi

AU - Kurumizaka, Hitoshi

AU - Kagawa, Wataru

PY - 2016

Y1 - 2016

N2 - The Rad52 protein is a eukaryotic single-strand DNA-annealing protein that is involved in the homologous recombinational repair of DNA double-strand breaks. The isolated N-terminal half of the human RAD52 protein (RAD521-212) forms an undecameric ring structure with a surface that is mostly positively charged. In the present study, it was found that RAD521-212 containing alanine mutations of the charged surface residues (Lys102, Lys133 and Glu202) is highly amenable to crystallization. The structure of the mutant RAD521-212 was solved at 2.4 Å resolution. The structure revealed an association between the symmetry-related RAD521-212 rings, in which a partially unfolded, C-terminal region of RAD52 extended into the DNA-binding groove of the neighbouring ring in the crystal. The alanine mutations probably reduced the surface entropy of the RAD521-212 ring and stabilized the ring-ring association observed in the crystal.

AB - The Rad52 protein is a eukaryotic single-strand DNA-annealing protein that is involved in the homologous recombinational repair of DNA double-strand breaks. The isolated N-terminal half of the human RAD52 protein (RAD521-212) forms an undecameric ring structure with a surface that is mostly positively charged. In the present study, it was found that RAD521-212 containing alanine mutations of the charged surface residues (Lys102, Lys133 and Glu202) is highly amenable to crystallization. The structure of the mutant RAD521-212 was solved at 2.4 Å resolution. The structure revealed an association between the symmetry-related RAD521-212 rings, in which a partially unfolded, C-terminal region of RAD52 extended into the DNA-binding groove of the neighbouring ring in the crystal. The alanine mutations probably reduced the surface entropy of the RAD521-212 ring and stabilized the ring-ring association observed in the crystal.

KW - higher order interaction

KW - homologous recombinational repair

KW - RAD52

KW - single-strand annealing proteins

KW - ssDNA-binding protein

KW - surface-entropy reduction

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U2 - 10.1107/S2053230X1601027X

DO - 10.1107/S2053230X1601027X

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C2 - 27487923

AN - SCOPUS:84982943392

SN - 1744-3091

VL - 72

SP - 598

EP - 603

JO - Acta Crystallographica Section F:Structural Biology Communications

JF - Acta Crystallographica Section F:Structural Biology Communications

ER -

Structure of the human DNA-repair protein RAD52 containing surface mutations

Abstract

Keywords

ASJC Scopus subject areas

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