Regulation of double-strand break-induced mammalian homologous recombination by UBL1, a RAD51-interacting protein

Li, Wenhui; Hesabi, Bahar; Babbo, Angela; Pacione, Carol; Liu, Jingmei; Chen, David J.; Nickoloff, Jac A.; Shen, Zhiyuan

doi:10.1093/nar/28.5.1145

Cited by 65 publications

(52 citation statements)

References 90 publications

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“…In most instances, the biological significance of the SUMO-1 modification of these proteins is not clear. Both RAD51 and RAD52, which play an important role in homologous recombination repair of DNA damage, bind to SUMO-1 in a noncovalent manner, although, again, the significance of these interactions remains unclear (47). Proteins that contribute to homologous recombination repair, such as BLM, RAD51, and the Mre11-RAD51-NBS1 complex, also colocalize with PML in NBs in response to the induction of DNA double-strand breaks by ionizing radiation (22,48,49).…”

Section: Discussionmentioning

confidence: 99%

Noncovalent SUMO-1 Binding Activity of Thymine DNA Glycosylase (TDG) Is Required for Its SUMO-1 Modification and Colocalization with the Promyelocytic Leukemia Protein

Takahashi

Hatakeyama

Saitoh

et al. 2005

Journal of Biological Chemistry

102

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SUMO-1 is a member of a family of ubiquitin-like molecules that are post-translationally conjugated to various cellular proteins in a process that is mechanistically similar to ubiquitylation. To identify molecules that bind noncovalently to SUMO-1, we performed yeast twohybrid screening with a SUMO-1 mutant that cannot be conjugated to target proteins as the bait. This screening resulted in the isolation of cDNAs encoding the b isoform of thymine DNA glycosylase (TDGb). A deletion mutant of TDGb (TDGb(⌬11)) that lacks a region shown to be required for noncovalent binding of SUMO-1 was also found not to be susceptible to SUMO-1 conjugation at an adjacent lysine residue, suggesting that such binding is required for covalent modification. In contrast, another mutant of TDGb (TDGb(KR)) in which the lysine residue targeted for SUMO-1 conjugation is replaced with arginine retained the ability to bind SUMO-1 noncovalently. TDGb was shown to interact with the promyelocytic leukemia protein (PML) in vitro as well as to colocalize with this protein to nuclear bodies in transfected cells. TDGb(KR) also colocalized with PML, whereas TDGb(⌬11) did not, indicating that the noncovalent SUMO-1 binding activity of TDGb is required for colocalization with PML. Furthermore, SUMO-1 modification of TDGb and PML enhanced the interaction between the two proteins. These results suggest that SUMO-1 functions to tether proteins to PML-containing nuclear bodies through post-translational modification and noncovalent protein-protein interaction.Post-translational modification of proteins plays important roles in the regulation of protein function and localization. Proteins are chemically modified by various molecules, including phosphate, lipids, and sugars. Modification by ubiquitin is distinct in that the modifier itself is a small protein. Ubiquitin is usually attached to a lysine residue of target proteins, resulting in the formation of a branched isopeptide chain. Such ubiquitylation serves to mark proteins for degradation by the 26 S proteasome.SUMO-1 is a member of the ubiquitin-like protein superfamily and is post-translationally conjugated to various cellular proteins in a process that is mechanistically analogous to ubiquitylation. SUMO-1 modification is mediated by a SUMO-1-activating enzyme (E1), 1 a SUMO-1-conjugating enzyme (E2), and a SUMO-1 ligase (E3) (1, 2). SUMO-1 is attached to target proteins via an isopeptide bond between the COOH-terminal glycine residue of SUMO-1 and the ⑀-amino group of a target lysine residue. A single E1 enzyme (the Aos1-Uba2 heterodimer) and a single E2 enzyme (Ubc9) have been identified for the SUMO-1 modification pathway in yeast and higher eukaryotes (2, 3). These two enzymes are sufficient to modify various SUMO-1 targets, including IB␣ (4), RanGAP1 (5), and p53 (6) in vitro, and it had been thought that SUMO-1 modification does not require an E3 ligase. However, several E3-like factors (PIAS family, RanBP2, PC2) for SUMO-1 modification were recently identified in yeast and mammalian cells ...

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Section: Discussionmentioning

confidence: 99%

Noncovalent SUMO-1 Binding Activity of Thymine DNA Glycosylase (TDG) Is Required for Its SUMO-1 Modification and Colocalization with the Promyelocytic Leukemia Protein

Takahashi

Hatakeyama

Saitoh

et al. 2005

Journal of Biological Chemistry

102

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“…The two key components in the early steps of HR the RecA recombinase Rad51 and the assembly factor Rad52 are candidate SUMO-target proteins, since they interact with both Ubc9 and SUMO in the yeast-two hybrid system (Shen et al, 1996). Although covalent modification of mammalian Rad51/52 has not yet been shown, SUMO was found in a complex with Rad51/Rad52 in mammalian cells (Li et al, 2000). Furthermore, the orthologue of RAD52 in fission yeast S. pombe is covalently modified by SUMO in a reconstituted in vitro system (Ho et al, 2001).…”

Section: Sumo In Replication and Homologous Recombinationmentioning

confidence: 99%

SUMO: a regulator of gene expression and genome integrity

2004

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Post-translational modification with the ubiquitin-like SUMO protein is involved in the regulation of many cellular key processes. The SUMO system modulates signal transduction pathways, including cytokine, Wnt, growth factor and steroid hormone signalling. SUMO frequently restrains the activity of downstream transcription factors in these pathways presumably by facilitating the recruitment of corepressors or mediating the assembly of repressor complexes. Additionally, evidence is accumulating that SUMO controls pathways important for the surveillance of genome integrity. SUMO regulates the PML/p53 tumour suppressor network, a key determinant in the cellular response to DNA damage. Moreover, proteins that maintain genomic stability by functioning at the interface between DNA replication, recombination and repair processes undergo SUMOylation. We will discuss some key findings that exemplify the role of SUMO in transcriptional regulation and genome surveillance.

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“…His-tagged SUMO-1 and UBC9 proteins were puri®ed from E. coli (BC21/DE3) carrying expression plasmid pETH-28 (Li et al, 2000) by using Ni2 + Kit (Novagen). Total HeLa cell extract were obtained by lysing 4610 7 cells in 1 ml of lysis bu er (20 mM Tris HCl pH 7.5, 0.5 mM DTT, 5 mM NaF, 1 mM Na Orthovanadate, 1 mM PMSF, 5 mg/ml leupeptin, 10 mg/ml aprotinin) and incubated on ice for 15 min (Desterro et al, 1997).…”

Section: In Vitro Sumo-1 Conjugation Assaymentioning

confidence: 99%

Functional analysis and intracellular localization of p53 modified by SUMO-1

et al. 2001

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p53 tumor suppressor is a subject of several posttranslational modi®cations, including phosphorylation, ubiquitination and acetylation, which regulate p53 function. A new covalent modi®cation of p53 at lysine 386 by SUMO-1 was recently identi®ed. To elucidate the function of sumoylated p53, we compared the properties of wild type p53 and sumoylation-de®cient p53 mutant, K386R. No di erences were found between wild type p53 and K386R mutant of p53 in transactivation or growth suppression assays. Moreover, overexpression of SUMO-1 has no e ect on p53-regulated transcription. Biochemical fractionation showed that sumoylated p53 is localized in the nucleus and is tightly bound to chromatin structures. p53 and SUMO-1 colocalized in PML nuclear bodies in 293 cells and the nucleoli in MCF7 and HT1080 cells. However, sumoylation-de®cient p53 mutant showed a similar pattern of intranuclear localization, suggesting that SUMO-1 does not target p53 to subnuclear structures. These data indicate that SUMO-1 modi®cation of p53 at lysine 386 may not be essential for p53's cellular localization, transcriptional activation, or growth regulation. Oncogene (2001) 20, 2587 ± 2599.

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Regulation of double-strand break-induced mammalian homologous recombination by UBL1, a RAD51-interacting protein

Cited by 65 publications

References 90 publications

Noncovalent SUMO-1 Binding Activity of Thymine DNA Glycosylase (TDG) Is Required for Its SUMO-1 Modification and Colocalization with the Promyelocytic Leukemia Protein

Noncovalent SUMO-1 Binding Activity of Thymine DNA Glycosylase (TDG) Is Required for Its SUMO-1 Modification and Colocalization with the Promyelocytic Leukemia Protein

SUMO: a regulator of gene expression and genome integrity

Functional analysis and intracellular localization of p53 modified by SUMO-1

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