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

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 RAD521–212containing alanine mutations of the charged surface residues (Lys102, Lys133 and Glu202) is highly amenable to crystallization. The structure of the mutant RAD521–212was solved at 2.4 Å resolution. The structure revealed an association between the symmetry-related RAD521–212rings, 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 RAD521–212ring and stabilized the ring–ring association observed in the crystal.

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Investigating SLX4‐Dependent Recruitment of a DNA Repair Protein to DNA Double Strand Breaks in Dividing Cells

The FASEB Journal ◽

10.1096/fasebj.2020.34.s1.08930 ◽

2020 ◽

Vol 34 (S1) ◽

pp. 1-1

Author(s):

Jonathan Hakimian ◽

Gyana Cureg ◽

Paula Fischhaber

Keyword(s):

Dna Repair ◽

Double Strand Breaks ◽

Dna Double Strand Breaks ◽

Strand Breaks ◽

Repair Protein ◽

Dividing Cells ◽

Dna Repair Protein

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Role of the yeast DNA repair protein Nej1 in end processing during the repair of DNA double strand breaks by non-homologous end joining

DNA Repair ◽

10.1016/j.dnarep.2015.04.003 ◽

2015 ◽

Vol 31 ◽

pp. 1-10 ◽

Cited By ~ 8

Author(s):

Hui Yang ◽

Yoshihiro Matsumoto ◽

Kelly M. Trujillo ◽

Susan P. Lees-Miller ◽

Mary Ann Osley ◽

...

Keyword(s):

Dna Repair ◽

Double Strand Breaks ◽

Dna Double Strand Breaks ◽

End Joining ◽

Strand Breaks ◽

Repair Protein ◽

Dna Repair Protein ◽

Non Homologous End Joining

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DNA double-strand breaks in heterochromatin elicit fast repair protein recruitment, histone H2AX phosphorylation and relocation to euchromatin

Nucleic Acids Research ◽

10.1093/nar/gkr230 ◽

2011 ◽

Vol 39 (15) ◽

pp. 6489-6499 ◽

Cited By ~ 188

Author(s):

Burkhard Jakob ◽

Jörn Splinter ◽

Sandro Conrad ◽

Kay-Obbe Voss ◽

Daniele Zink ◽

...

Keyword(s):

Double Strand Breaks ◽

Dna Double Strand Breaks ◽

H2ax Phosphorylation ◽

Strand Breaks ◽

Histone H2ax ◽

Repair Protein ◽

Fast Repair ◽

Protein Recruitment ◽

Histone H2ax Phosphorylation

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Spatiotemporal dynamics of 53BP1 dimer recruitment to a DNA double strand break

Nature Communications ◽

10.1038/s41467-020-19504-3 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Jieqiong Lou ◽

David G. Priest ◽

Ashleigh Solano ◽

Adèle Kerjouan ◽

Elizabeth Hinde

Keyword(s):

Nuclear Architecture ◽

Strand Break ◽

Cell System ◽

Spatiotemporal Dynamics ◽

Dna Double Strand Breaks ◽

Strand Breaks ◽

Dna Double Strand Break ◽

Repair Protein ◽

Dna Repair Protein ◽

Self Association

AbstractTumor suppressor p53-binding protein 1 (53BP1) is a DNA repair protein essential for the detection, assessment, and resolution of DNA double strand breaks (DSBs). The presence of a DSB is signaled to 53BP1 via a local histone modification cascade that triggers the binding of 53BP1 dimers to chromatin flanking this type of lesion. While biochemical studies have established that 53BP1 exists as a dimer, it has never been shown in a living cell when or where 53BP1 dimerizes upon recruitment to a DSB site, or upon arrival at this nuclear location, how the DSB histone code to which 53BP1 dimers bind regulates retention and self-association into higher-order oligomers. Thus, here in live-cell nuclear architecture we quantify the spatiotemporal dynamics of 53BP1 oligomerization during a DSB DNA damage response by coupling fluorescence fluctuation spectroscopy (FFS) with the DSB inducible via AsiSI cell system (DIvA). From adopting this multiplexed approach, we find that preformed 53BP1 dimers relocate from the nucleoplasm to DSB sites, where consecutive recognition of ubiquitinated lysine 15 of histone 2A (H2AK15ub) and di-methylated lysine 20 of histone 4 (H4K20me2), leads to the assembly of 53BP1 oligomers and a mature 53BP1 foci structure.

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Induction of Biphasic DNA Double Strand Breaks and Activation of Multiple Repair Protein Complexes by DNA Topoisomerase I Drug 7-Ethyl-10-hydroxy-camptothecin

Molecular Pharmacology ◽

10.1124/mol.61.4.742 ◽

2002 ◽

Vol 61 (4) ◽

pp. 742-748 ◽

Cited By ~ 27

Author(s):

Jiaxi Wu ◽

Ming-biao Yin ◽

Gunnar Hapke ◽

Károly Tóth ◽

Youcef M. Rustum

Keyword(s):

Topoisomerase I ◽

Protein Complexes ◽

Double Strand Breaks ◽

Dna Topoisomerase I ◽

Dna Double Strand Breaks ◽

Dna Topoisomerase ◽

Strand Breaks ◽

Repair Protein

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RecO impedes RecG-SSB binding to impair the strand annealing recombination pathway in E.coli

10.1101/708271 ◽

2019 ◽

Author(s):

Xuefeng Pan ◽

Li Yang ◽

Nan Jiang ◽

Xifang Chen ◽

Bo Li ◽

...

Keyword(s):

Escherichia Coli ◽

Dna Repair ◽

Dna Replication ◽

Replication Fork ◽

Double Strand Breaks ◽

Recombinational Repair ◽

Dna Double Strand Breaks ◽

Strand Breaks ◽

Homologous Recombinational Repair ◽

Differential Binding

AbstractFaithful duplication of genomic DNA relies not only on the fidelity of DNA replication itself, but also on fully functional DNA repair and homologous recombination machinery. We report a molecular mechanism responsible for deciding homologous recombinational repair pathways during replication dictated by binding of RecO and RecG to SSB in E.coli. Using a RecG-yfp fusion protein, we found that RecG-yfp foci appeared only in the ΔrecG, ΔrecO and ΔrecA, ΔrecO double mutants. Surprisingly, foci were not observed in wild-type ΔrecG, or double mutants where recG and either recF or, separately recR were deleted. In addition, formation of RecG-yfp foci in the ΔrecO::kanR required wildtype ssb, as ssb-113 could not substitute. This suggests that RecG and RecO binding to SSB is competitive. We also found that the UV resistance of recO alone mutant increased to certain extent by supplementing RecG. In an ssb-113 mutant, RecO and RecG worked following a different pattern. Both RecO and RecG were able to participate in repairing UV damages when grown at permissive temperature, while they could also be involved in making DNA double strand breaks when grown at nonpermissive temperature. So, our results suggested that differential binding of RecG and RecO to SSB in a DNA replication fork in Escherichia coli.may be involved in determining whether the SDSA or DSBR pathway of homologous recombinational repair is used.Author summarySingle strand DNA binding proteins (SSB) stabilize DNA holoenzyme and prevent single strand DNA from folding into non-B DNA structures in a DNA replication fork. It has also been revealed that SSB can also act as a platform for some proteins working in DNA repair and recombination to access DNA molecules when DNA replication fork needs to be reestablished. In Escherichia coli, several proteins working primarily in DNA repair and recombination were found to participate in DNA replication fork resumption by physically interacting with SSB, including RecO and RecG etc. However the hierarchy of these proteins interacting with SSB in Escherichia coli has not been well defined. In this study, we demonstrated a differential binding of RecO and RecG to SSB in DNA replication was used to establish a RecO-dependent pathway of replication fork repair by abolishing a RecG-dependent replication fork repair. We also show that, RecG and RecO could randomly participate in DNA replication repair in the absence of a functional SSB, which may be responsible for the generation of DNA double strand breaks in an ssb-113 mutant in Escherichia coli.

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