Rap80 Protein Recruitment to DNA Double-strand Breaks Requires Binding to Both Small Ubiquitin-like Modifier (SUMO) and Ubiquitin Conjugates

Hu, Xin; Paul, Atanu; Wang, Bin

doi:10.1074/jbc.m112.374116

Cited by 67 publications

(85 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Hybrid SUMO-ubiquitin chains that consist of a SUMO2 chain joined to a K63-linked ubiquitin chain have been described (Tatham et al, 2013), and can be specifically recognized by some proteins (Geoffroy and Hay, 2009;Guzzo et al, 2012;Hu et al, 2012). The UMI and/or MUI1 sequences of RNF168 that we found to be important for recovery of SUMO2 chains have been previously shown to recognize K63-linked ubiquitin (Panier et al, 2012), raising the possibility that the SUMO2-containing material that we recovered are hybrid SUMO-ubiquitin chains.…”

Section: Rnf168 Associates With Hybrid Sumo2-ubiquitin Chainsmentioning

confidence: 80%

See 1 more Smart Citation

Identification of RNF168 as a PML nuclear body regulator

Shire

Wong

Tatham

et al. 2016

Journal of Cell Science

View full text Add to dashboard Cite

Promyelocytic leukemia (PML) protein forms the basis of PML nuclear bodies (PML NBs), which control many important processes. We have screened an shRNA library targeting ubiquitin pathway proteins for effects on PML NBs, and identified RNF8 and RNF168 DNAdamage response proteins as negative regulators of PML NBs. Additional studies confirmed that depletion of either RNF8 or RNF168 increased the levels of PML NBs and proteins, whereas overexpression induced loss of PML NBs. RNF168 partially localized to PML NBs through its UMI/MIU1 ubiquitin-interacting region and associated with NBs formed by any PML isoform. The association of RNF168 with PML NBs resulted in increased ubiquitylation and SUMO2 modification of PML. In addition, RNF168 was found to associate with proteins modified by SUMO2 and/or SUMO3 in a manner dependent on its ubiquitin-binding sequences, suggesting that hybrid SUMO-ubiquitin chains can be bound. In vitro assays confirmed that RNF168, preferentially, binds hybrid SUMO2-K63 ubiquitin chains compared with K63-ubiquitin chains or individual SUMO2. Our study identified previously unrecognized roles for RNF8 and RNF168 in the regulation of PML, and a so far unknown preference of RNF168 for hybrid SUMOubiquitin chains.

show abstract

Section: Rnf168 Associates With Hybrid Sumo2-ubiquitin Chainsmentioning

confidence: 80%

“…We now suggest that this requirement is due to the generation of hybrid SUMO-ubiquitin chains, which are then recognized by RNF168. This would be similar to the recruitment of RAP80 to double-stranded DNA breaks, which has been shown to involve binding to SUMO-ubiquitin hybrid chains generated by RNF4 (Hu et al, 2012;Guzzo et al, 2012).…”

Section: Discussionmentioning

confidence: 99%

Identification of RNF168 as a PML nuclear body regulator

Shire

Wong

Tatham

et al. 2016

Journal of Cell Science

View full text Add to dashboard Cite

show abstract

“…Recruitment of RAP80 to IRIF is 53BP1-independent, but dependent on hybrid ubiquitin-SUMO chains that are recognized by ubiquitin and SUMO-interacting motifs present within RAP80. 51,52 RAP80 facilitates the subsequent recruitment of BRCA1 and repair of DNA DSBs by homologous recombination. [53][54][55][56] Treatment of U2OS cells with MG132 inhibited the formation of RAP80 IRIF (Fig.…”

Section: Expression Of H2ax Fusion Proteins Rescues 53bp1 Irif In Celmentioning

confidence: 99%

Ubiquitin-H2AX fusions render 53BP1 recruitment to DNA damage sites independent of RNF8 or RNF168

et al. 2015

View full text Add to dashboard Cite

The mammalian E3 ubiquitin ligases RNF8 and RNF168 facilitate recruitment of the DNA damage response protein 53BP1 to sites of DNA double-strand breaks (DSBs). The mechanism involves recruitment of RNF8, followed by recruitment of RNF168, which ubiquitinates histones H2A/H2AX on K15. 53BP1 then binds to nucleosomes at sites of DNA DSBs by recognizing, in addition to methyl marks, histone H2A/H2AX ubiquitinated on K15. We report here that expressing H2AX fusion proteins with N-terminal bulky moieties can rescue 53BP1 recruitment to sites of DNA DSBs in cells lacking RNF8 or RNF168 or in cells treated with proteasome inhibitors, in which histone ubiquitination at sites of DNA DSBs is compromised. The rescue required S139 at the C-terminus of the H2AX fusion protein and was occasionally accompanied by partial rescue of ubiquitination at sites of DNA DSBs. We conclude that recruitment of 53BP1 to sites of DNA DSBs is possible in the absence of RNF8 or RNF168, but still dependent on chromatin ubiquitination.

show abstract

“…The accumulation of SUMO at sites of DNA damage is important for the efficient recruitment of DDR proteins such as BRCA1 and 53BP1 Guzzo et al 2012;Hu et al 2012). Furthermore, a group of proteins involved in homologous recombination (HR), including BRCA1, MDC1, RPA70, and BLM, is directly modified by SUMO (Morris et al 2009;Ouyang et al 2009b;Dou et al 2010;Galanty et al 2012;Luo et al 2012;Yin et al 2012).…”

mentioning

confidence: 99%

SUMOylation of ATRIP potentiates DNA damage signaling by boosting multiple protein interactions in the ATR pathway

Ouyang

Mori

et al. 2014

Genes Dev.

View full text Add to dashboard Cite

The ATR (ATM [ataxia telangiectasia-mutated]-and Rad3-related) checkpoint is a crucial DNA damage signaling pathway. While the ATR pathway is known to transmit DNA damage signals through the ATR-Chk1 kinase cascade, whether post-translational modifications other than phosphorylation are important for this pathway remains largely unknown. Here, we show that protein SUMOylation plays a key role in the ATR pathway. ATRIP, the regulatory partner of ATR, is modified by SUMO2/3 at K234 and K289. An ATRIP mutant lacking the SUMOylation sites fails to localize to DNA damage and support ATR activation efficiently. Surprisingly, the ATRIP SUMOylation mutant is compromised in the interaction with a protein group, rather than a single protein, in the ATR pathway. Multiple ATRIP-interacting proteins, including ATR, RPA70, TopBP1, and the MRE11-RAD50-NBS1 complex, exhibit reduced binding to the ATRIP SUMOylation mutant in cells and display affinity for SUMO2 chains in vitro, suggesting that they bind not only ATRIP but also SUMO. Fusion of a SUMO2 chain to the ATRIP SUMOylation mutant enhances its interaction with the protein group and partially suppresses its localization and functional defects, revealing that ATRIP SUMOylation promotes ATR activation by providing a unique type of protein glue that boosts multiple protein interactions along the ATR pathway.[Keywords: ATR; ATRIP; checkpoint; SUMOylation] Supplemental material is available for this article.Received January 18, 2014; revised version accepted June 2, 2014.The maintenance of genomic stability requires not only DNA repair machineries but also signal transduction pathways that regulate and coordinate the DNA damage response (DDR) (Ciccia and Elledge 2010). In human cells, DNA damage signaling is primarily initiated by the ataxia telangiectasia-mutated (ATM) and the ATMand Rad3-related (ATR) kinases. Whereas ATM is activated by DNA double-stranded breaks (DSBs), ATR is elicited by a much broader spectrum of DNA damage and replication stress (Cimprich and Cortez 2008;Marechal and Zou 2013;Shiloh and Ziv 2013). Once activated, ATM and ATR phosphorylate and activate their effector kinases, Chk2 and Chk1, respectively. Together, the ATMChk2 and ATR-Chk1 kinase cascades phosphorylate a number of substrates involved in DNA repair, DNA replication, and cell cycle transitions, coordinating these processes to suppress genomic instability. In addition to the phosphorylation events mediated by the ATM and ATR pathways, several other types of post-translational modifications (PTMs), such as ubiquitylation, SUMOylation, methylation, acetylation, and poly-ADP ribosylation, are also implicated in the DDR (Bekker-Jensen and Mailand 2010;Huen et al. 2010;Lukas et al. 2011;Jackson and Durocher 2013). We recently found that the efficient activation of ATR relies on a ubiquitylation circuitry mediated by RPA-ssDNA (RPA-coated ssDNA) and PRP19 (Marechal et al. 2014). This new finding raises a question as to whether other PTMs also participate in DNA damage signaling through the ATR path...

show abstract

Rap80 Protein Recruitment to DNA Double-strand Breaks Requires Binding to Both Small Ubiquitin-like Modifier (SUMO) and Ubiquitin Conjugates

Cited by 67 publications

References 32 publications

Identification of RNF168 as a PML nuclear body regulator

Identification of RNF168 as a PML nuclear body regulator

Ubiquitin-H2AX fusions render 53BP1 recruitment to DNA damage sites independent of RNF8 or RNF168

SUMOylation of ATRIP potentiates DNA damage signaling by boosting multiple protein interactions in the ATR pathway

Contact Info

Product

Resources

About