scholarly journals Noncanonical E2 Variant-Independent Function of UBC13 in Promoting Checkpoint Protein Assembly

2008 ◽  
Vol 28 (19) ◽  
pp. 6104-6112 ◽  
Author(s):  
Michael S. Y. Huen ◽  
Jun Huang ◽  
Jingsong Yuan ◽  
Masahiro Yamamoto ◽  
Shizuo Akira ◽  
...  
Keyword(s):  
Dna Damage ◽  
Ubiquitin Ligase ◽  
E3 Ubiquitin Ligase ◽  
Protein Assembly ◽  
Ring Domain ◽  
Dna Breaks ◽  
Focus Formation ◽  
Checkpoint Protein ◽  
Ubiquitin Ligase Complex ◽  
Ubiquitin Conjugating Enzyme

ABSTRACT The E2 ubiquitin-conjugating enzyme UBC13 plays pivotal roles in diverse biological processes. Recent studies have elucidated that UBC13, in concert with the E3 ubiquitin ligase RNF8, propagates the DNA damage signal via a ubiquitylation-dependent signaling pathway. However, mechanistically how UBC13 mediates its role in promoting checkpoint protein assembly and its genetic requirement for E2 variants remain elusive. Here we provide evidence to support the idea that the E3 ubiquitin ligase complex RNF8-UBC13 functions independently of E2 variants and is sufficient in facilitating ubiquitin conjugations and accumulation of DNA damage mediator 53BP1 at DNA breaks. The RNF8 RING domain serves as the molecular platform to anchor UBC13 at the damaged chromatin, where localized ubiquitylation events allow sustained accumulation of checkpoint proteins. Intriguingly, we found that only a group of RING domains derived from E3 ubiquitin ligases, which have been shown to interact with UBC13, enabled UBC13-mediated FK2 and 53BP1 focus formation at DNA breaks. We propose that the RNF8 RING domain selects and loads a subset of UBC13 molecules, distinct from those that exist as heterodimers, onto sites of double-strand breaks, which facilitates the amplification of DNA damage signals.

DNA Damage-Induced Foci of E2 Ubiquitin-Conjugating Enzyme are Detectable upon Co-transfection with an Interacting E3 Ubiquitin Ligase

2015 ◽  
Vol 54 (2) ◽  
pp. 147-157 ◽  
Author(s):  
Degui Wang ◽  
Yingxia Tian ◽  
Dong Wei ◽  
Yuhong Jing ◽  
Haitao Niu ◽  
...  
Keyword(s):  
Dna Damage ◽  
Ubiquitin Ligase ◽  
E3 Ubiquitin Ligase ◽  
Ubiquitin Conjugating Enzyme ◽  
Conjugating Enzyme

scholarly journals RNF168-mediated localization of BARD1 recruits the BRCA1-PALB2 complex to DNA damage

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
John J. Krais ◽  
Yifan Wang ◽  
Pooja Patel ◽  
Jayati Basu ◽  
Andrea J. Bernhardy ◽  
...  
Keyword(s):  
Dna Damage ◽  
Ubiquitin Ligase ◽  
E3 Ubiquitin Ligase ◽  
Coiled Coil ◽  
Histone H2a ◽  
Dna Breaks ◽  
Diverse Range ◽  
Dna Damage Signaling ◽  
Dna Repair Proteins ◽  
K 13

AbstractDNA damage prompts a diverse range of alterations to the chromatin landscape. The RNF168 E3 ubiquitin ligase catalyzes the mono-ubiquitination of histone H2A at lysine (K)13/15 (mUb-H2A), forming a binding module for DNA repair proteins. BRCA1 promotes homologous recombination (HR), in part, through its interaction with PALB2, and the formation of a larger BRCA1-PALB2-BRCA2-RAD51 (BRCA1-P) complex. The mechanism by which BRCA1-P is recruited to chromatin surrounding DNA breaks is unclear. In this study, we reveal that an RNF168-governed signaling pathway is responsible for localizing the BRCA1-P complex to DNA damage. Using mice harboring a Brca1CC (coiled coil) mutation that blocks the Brca1-Palb2 interaction, we uncovered an epistatic relationship between Rnf168− and Brca1CC alleles, which disrupted development, and reduced the efficiency of Palb2-Rad51 localization. Mechanistically, we show that RNF168-generated mUb-H2A recruits BARD1 through a BRCT domain ubiquitin-dependent recruitment motif (BUDR). Subsequently, BARD1-BRCA1 accumulate PALB2-RAD51 at DNA breaks via the CC domain-mediated BRCA1-PALB2 interaction. Together, these findings establish a series of molecular interactions that connect the DNA damage signaling and HR repair machinery.

scholarly journals In Vitro Enhanced Sensitivity to Cisplatin in D67Y BRCA1 RING Domain Protein

2011 ◽  
Vol 5 ◽  
pp. BCBCR.S8184 ◽  
Author(s):  
Apichart Atipairin ◽  
Adisorn Ratanaphan
Keyword(s):  
Dna Damage ◽  
Ubiquitin Ligase ◽  
Dna Damage Repair ◽  
E3 Ubiquitin Ligase ◽  
Chemotherapeutic Agents ◽  
Ring Domain ◽  
Protein Ubiquitination ◽  
Damage Repair ◽  
Domain Protein

BRCA1 is a tumor suppressor protein involved in maintaining genomic integrity through multiple functions in DNA damage repair, transcriptional regulation, cell cycle checkpoint, and protein ubiquitination. The BRCA1-BARD1 RING complex has an E3 ubiquitin ligase function that plays essential roles in response to DNA damage repair. BRCA1-associated cancers have been shown to confer a hypersensitivity to chemotherapeutic agents. Here, we have studied the functional consequence of the in vitro E3 ubiquitin ligase activity and cisplatin sensitivity of the missense mutation D67Y BRCA1 RING domain. The D67Y BRCA1 RING domain protein exhibited the reduced ubiquitination function, and was more susceptible to the drug than the D67E or wild-type BRCA1 RING domain protein. This evidence emphasized the potential of using the BRCA1 dysfunction as an important determinant of chemotherapy responses in breast cancer.

scholarly journals RNF168-mediated localization of BARD1 recruits the BRCA1-PALB2 complex to DNA damage

2021 ◽  
Author(s):  
John J Krais ◽  
Yifan Wang ◽  
Pooja Patel ◽  
Jayati Basu ◽  
Andrea J Bernhardy ◽  
...  
Keyword(s):  
Dna Damage ◽  
Ubiquitin Ligase ◽  
E3 Ubiquitin Ligase ◽  
Coiled Coil ◽  
Histone H2a ◽  
Dna Breaks ◽  
Diverse Range ◽  
Dna Damage Signaling ◽  
Dna Repair Proteins ◽  
K 13

DNA damage prompts a diverse range of alterations to the chromatin landscape. The RNF168 E3 ubiquitin ligase catalyzes the mono-ubiquitination of histone H2A at lysine (K)13/15 (mUb-H2A), forming a binding module for DNA repair proteins. BRCA1 promotes homologous recombination (HR), in part, through its interaction with PALB2, and the formation of a larger BRCA1-PALB2-BRCA2-RAD51 (BRCA1-P) complex. The mechanism by which BRCA1-P is recruited to chromatin surrounding DNA breaks is unclear. In this study, we reveal that an RNF168-governed signaling pathway is responsible for localizing the BRCA1-P complex to DNA damage. Using mice harboring a Brca1CC (coiled coil) mutation that blocks the Brca1-Palb2 interaction, we uncovered an epistatic relationship between Rnf168- and Brca1CC alleles, which disrupted development, and reduced the efficiency of Palb2-Rad51 localization. Mechanistically, we show that RNF168-generated mUb-H2A recruits BARD1 through a BRCT domain ubiquitin-dependent recruitment motif (BUDR). Subsequently, BARD1-BRCA1 accumulate PALB2-RAD51 at DNA breaks via the CC domain-mediated BRCA1-PALB2 interaction. Together, these findings establish a series of molecular interactions that connect the DNA damage signaling and HR repair machinery.

scholarly journals UBE2T, the Fanconi Anemia Core Complex, and FANCD2 Are Recruited Independently to Chromatin: a Basis for the Regulation of FANCD2 Monoubiquitination

2007 ◽  
Vol 27 (24) ◽  
pp. 8421-8430 ◽  
Author(s):  
Arno Alpi ◽  
Frederic Langevin ◽  
Georgina Mosedale ◽  
Yuichi J. Machida ◽  
Anindya Dutta ◽  
...  
Keyword(s):  
Dna Damage ◽  
Ubiquitin Ligase ◽  
Fanconi Anemia ◽  
E3 Ubiquitin Ligase ◽  
S Phase ◽  
Multiprotein Complex ◽  
Core Complex ◽  
Nuclear Core ◽  
Ubiquitin Conjugating Enzyme ◽  
Fancd2 Monoubiquitination

ABSTRACT The Fanconi anemia (FA) nuclear core complex and the E2 ubiquitin-conjugating enzyme UBE2T are required for the S phase and DNA damage-restricted monoubiquitination of FANCD2. This constitutes a key step in the FA tumor suppressor pathway, and much attention has been focused on the regulation at this point. Here, we address the importance of the assembly of the FA core complex and the subcellular localization of UBE2T in the regulation of FANCD2 monoubiquitination. We establish three points. First, the stable assembly of the FA core complex can be dissociated of its ability to function as an E3 ubiquitin ligase. Second, the actual E3 ligase activity is not determined by the assembly of the FA core complex but rather by its DNA damage-induced localization to chromatin. Finally, UBE2T and FANCD2 access this subcellular fraction independently of the FA core complex. FANCD2 monoubiquitination is therefore not regulated by multiprotein complex assembly but by the formation of an active E2/E3 holoenzyme on chromatin.

scholarly journals Structure and function of E3 ubiquitin ligase mindbomb RING domain

2012 ◽  
Vol 26 (S1) ◽  
Author(s):  
Robert D Wardlow ◽  
Sung Hee Choi ◽  
Brian McMillan ◽  
Stephen C Blacklow
Keyword(s):  
Ubiquitin Ligase ◽  
E3 Ubiquitin Ligase ◽  
Structure And Function ◽  
Ring Domain ◽  
And Function

scholarly journals PARP1-dependent recruitment of the FBXL10-RNF68-RNF2 ubiquitin ligase to sites of DNA damage controls H2A.Z loading

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Gergely Rona ◽  
Domenico Roberti ◽  
Yandong Yin ◽  
Julia K Pagan ◽  
Harrison Homer ◽  
...  
Keyword(s):  
Dna Damage ◽  
Homologous Recombination ◽  
Ubiquitin Ligase ◽  
Transcriptional Repression ◽  
Genotoxic Stress ◽  
Strand Break ◽  
Homologous Recombination Repair ◽  
Dependent Manner ◽  
Ubiquitin Ligase Complex ◽  
Recombination Repair

The mammalian FBXL10-RNF68-RNF2 ubiquitin ligase complex (FRRUC) mono-ubiquitylates H2A at Lys119 to repress transcription in unstressed cells. We found that the FRRUC is rapidly and transiently recruited to sites of DNA damage in a PARP1- and TIMELESS-dependent manner to promote mono-ubiquitylation of H2A at Lys119, a local decrease of H2A levels, and an increase of H2A.Z incorporation. Both the FRRUC and H2A.Z promote transcriptional repression, double strand break signaling, and homologous recombination repair (HRR). All these events require both the presence and activity of the FRRUC. Moreover, the FRRUC and its activity are required for the proper recruitment of BMI1-RNF2 and MEL18-RNF2, two other ubiquitin ligases that mono-ubiquitylate Lys119 in H2A upon genotoxic stress. Notably, whereas H2A.Z is not required for H2A mono-ubiquitylation, impairment of the latter results in the inhibition of H2A.Z incorporation. We propose that the recruitment of the FRRUC represents an early and critical regulatory step in HRR.

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