Novel function of HATs and HDACs in homologous recombination through acetylation of human RAD52 at double-strand break sites

Yasuda, Takeshi; Kagawa, Wataru; Ogi, Tomoo; Kato, Takamitsu A.; Suzuki, Takehiro; Dohmae, Naoshi; Takizawa, Kazuya; Nakazawa, Yuka; Genet, Matthew; Saotome, Mika; Hama, Michio; Konishi, Teruaki; Nakajima, Naomasa; Hazawa, Masaharu; Tomita, Masaru; Koike, Manabu; Noshiro, Katsuko; Tomiyama, Katsunori; Obara, Chizuka; Gotoh, Tadao; Ui, Ayako; Fujimori, A.; Nakayama, Fumiaki; Hanaoka, Fumio; Sugasawa, Kaoru; Okayasu, Ryuichi; Jeggo, Penny A.; Tajima, Katsushi

doi:10.1371/journal.pgen.1007277

Cited by 29 publications

(39 citation statements)

References 50 publications

(75 reference statements)

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“…Since nonhistone acetylation was first identified, increasingly more functions for acetylation have been found to act on various life processes, including DNA damage response and autophagy (Botrugno et al, 2012;Zhong et al, 2017;Yasuda et al, 2018), genomic stability (Billon et al, 2017;Fournier and Tora, 2017), transcriptional activity (Seo et al, 2015), protein degradation (Liu et al, 2013;Wei et al, 2018) and lysosomal function (Zhang et al, 2018). Previous studies have shown that acetylation could compete with ubiquitination at the same lysine residue, thus blocking the ubiquitin-mediated proteasomal degradation pathway to improve the protein stability, which is involved in cell cycle regulation (Lahusen et al, 2018), tumour suppression and progression (Wan et al, 2015;Choi et al, 2017), bacterial virulence (Sang et al, 2017) and signal transduction (Garcia-Aguilar et al, 2016;Beckwith et al, 2018;Wei et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

The effect of acetylation on the protein stability of BmApoLp‐III in the silkworm, Bombyx mori

Yang

Zhu

Liu

et al. 2019

Insect Molecular Biology

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Acetylation is an important, reversible posttranslational modification to a protein. In a previous study, we found that there were a large number of acetylated sites in various nutrient storage proteins of the silkworm haemolymph. In this study, we confirmed that acetylation can affect the stability of nutrient storage protein Bombyx mori apolipophorin‐III (BmApoLp‐III). First, the expression of BmApoLp‐III could be upregulated when BmN cells were treated with the deacetylase inhibitor panobinostat (LBH589); similarly, the expression was downregulated when the cells were treated with the acetylase inhibitor C646. Furthermore, the increase in acetylation by LBH589 could inhibit the degradation and improve the accumulation of BmApoLp‐III in BmN cells treated with cycloheximide and MG132 respectively. Moreover, we found that an increase in acetylation could decrease the ubiquitination of BmApoLp‐III and vice versa; therefore, we predicted that acetylation could improve the stability of BmApoLp‐III by competing for ubiquitination and inhibiting the protein degradation pathway mediated by ubiquitin. Additionally, BmApoLp‐III had an antiapoptosis function that increased after LBH589 treatment, which might have been due to the improved protein stability after acetylation. These results have laid the foundation for further study on the mechanism of acetylation in regulating the storage and utilization of silkworm nutrition.

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Section: Discussionmentioning

confidence: 99%

The effect of acetylation on the protein stability of BmApoLp‐III in the silkworm, Bombyx mori

Yang

Zhu

Liu

et al. 2019

Insect Molecular Biology

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“…In the case of the insertion of a long DNA fragment (more than 1.5 kb), the knock-in efficiency increased as the length of the homology arms increased, up to about 1500 bp [48]. Thus, long homology arms (more than 500 bp) are usually used for the knock-in of such a long DNA fragment [45,[49][50][51][52]. In this case, the knock-in is mediated by HR.…”

Section: Donor Dna Plasmid For Homology Directed Gene Knock-inmentioning

confidence: 99%

“…In this case, the knock-in is mediated by HR. The knock-in efficiency by HR is low [53], and accordingly, a selectable marker is introduced into the donor DNA plasmid for screening the knock-in clones in some cases [49][50][51][52]. Therefore, the selectable donor DNA plasmid for the HR-mediated gene knock-in contains left and right arms, the inserted gene of interest, and a selectable marker gene with promoter and transcription terminator regions.…”

Section: Donor Dna Plasmid For Homology Directed Gene Knock-inmentioning

confidence: 99%

MultiSite Gateway Technology Is Useful for Donor DNA Plasmid Construction in CRISPR/Cas9-Mediated Knock-In System

Yasuda¹,

Tajima²

2019

Modulating Gene Expression - Abridging the RNAi and CRISPR-Cas9 Technologies

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The clustered regulatory interspaced short palindromic repeats (CRISPR)/Cas9 method is a powerful tool for genome editing, by introducing a DNA double-strand break (DSB) at the specific site. The gene knockout can be achieved by the deletion or insertion at the CRISPR/Cas9-mediated DSB site by error-prone nonhomologous end joining repair in targeted cells. However, the gene knock-in is still difficult as compared to the knockout , because of the low efficiency of homology directed repair with donor DNA in cells. Therefore, to efficiently select the knock-in cells, we developed a complicated donor DNA plasmid containing an antibiotic-resistance gene, in addition to the knock-in sequence and the two homology arms. MultiSite Gateway technology is a useful tool for constructing this complicated plasmid. We describe the MultiSite Gateway technology and provide an overview of the DSB repair pathways to clarify the knockout and knockin methods by the CRISPR/Cas9 system.

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“…Its RPA binding domains underlie RAD52´s biochemical functions in binding RPA-coated ssDNA, annealing, and homology-directed repair. Even though many details regarding the contribution of posttranslational modification to the function of RAD52 remain to be explored, we know that RAD52 acetylation is required for its accumulation at DSBs, sumoylation of yeast RAD52 for the choice of SSA over canonical recombination at repeats, and tyrosine phosphorylation of RAD52 for ssDNA, rather than dsDNA binding and the choice of SSA [6][7][8][9].…”

Section: Introductionmentioning

confidence: 99%

Current Understanding of RAD52 Functions: Fundamental and Therapeutic Insights

Gottifredi

Wiesmüller

2020

Cancers

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In this Special Issue, we would like to focus on the various functions of the RAD52 helicase-like protein and the current implications of such findings for cancer treatment. Over the last few years, various laboratories have discovered particular activities of mammalian RAD52-both in S and M phase-that are distinct from the auxiliary role of yeast RAD52 in homologous recombination. At DNA double-strand breaks, RAD52 was demonstrated to spur alternative pathways to compensate for the loss of homologous recombination functions. At collapsed replication forks, RAD52 activates break-induced replication. In the M phase, RAD52 promotes the finalization of DNA replication. Its compensatory role in the resolution of DNA double-strand breaks has put RAD52 in the focus of synthetic lethal strategies, which is particularly relevant for cancer treatment.Over the last years, new functions of RAD52 have been identified. For example, various laboratories have discovered the involvement of mammalian RAD52 and RNA templates and RNA-DNA hybrid structures like R loops in unprecedented homology-directed DSB repair events [9]. First, RNA was discovered to serve as a bridging template in RAD52-and RPA-mediated homology-directed DSB repair, which may play a role during transcription, replication, class-switch recombination, and at telomeres. Second, in transcription-coupled HR (TC-HR) in G0/G1 cells, R loops generated during transcription were found to be bound by CSB, followed by RAD52-mediated, BRCA-independent RAD51 recruitment, and HR. Third, in transcription-activated HR (TA-HR) in S/G2 cells, RAD52 recruits the XPG cleaving R loops, which generates ssDNA overhangs for BRCA-dependent HR [9].Surprisingly, RAD52 also acts during events other than canonical DSB repair-namely, during DNA replication when it counteracts excessive fork regression, which may exhaust protection factors, causing breakage of reversed forks. At collapsed replication forks, RAD52 activates break-induced replication (BIR), a specialized pathway that repairs single-ended DSBs. During M phase, RAD52-mediated BIR also promotes the finalization of DNA replication (MiDAS-mitotic DNA synthesis). Notably, RAD52 cooperates with various nucleases, namely, MUS81 during BIR, ERCC1/XPF during SSA, and XPG during TA-HR. It also cooperates with MRE11 and MUS81/EME1 at de-protected forks that have reverted to process these structures into HR substrates, ultimately enabling the cell to continue DNA replication [10][11][12]. The multiple activities of RAD52 known to date are summarized in Figure 1.Cancers 2020, 12, x FOR PEER REVIEW 2 of 8

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Novel function of HATs and HDACs in homologous recombination through acetylation of human RAD52 at double-strand break sites

Cited by 29 publications

References 50 publications

The effect of acetylation on the protein stability of BmApoLp‐III in the silkworm, Bombyx mori

The effect of acetylation on the protein stability of BmApoLp‐III in the silkworm, Bombyx mori

MultiSite Gateway Technology Is Useful for Donor DNA Plasmid Construction in CRISPR/Cas9-Mediated Knock-In System

Current Understanding of RAD52 Functions: Fundamental and Therapeutic Insights

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