IndraLab
Statements
reach
"Furthermore, by genetically tagging UAF1 with a fluorescent protein, such as RFP, and using automated microscopy procedures, the assay can be readily adapted to a high throughput format in order to allow a high content screening for molecules that interfere with USP1 and UAF1 complex formation."
sparser
"Indeed, the structure of USP1‐UAF1 complex bound to ID2 Ub ‐DNA revealed ID2 movements towards the open‐state conformation, affecting not only FANCI helices in the region where ubiquitin‐conjugation occurs, but also, and most profoundly, the FANCD2 N‐terminus involved in interaction with FANCI's ubiquitin (Rennie et al , xref )."
sparser
"However, while the mechanism of FANCD2 ubiquitination and deubiquitination has been sufficiently elucidated (Chaugule et al , xref ; Rennie et al , xref ; Wang et al , xref ), we are still lacking information on how UBE2T and FA‐core engage the mono‐ubiquitinated ID2 complex for FANCI ubiquitination, and how the ubiquitin from FANCI is removed by USP1‐UAF1."
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"Altogether, these results support an idea that USP1 and UAF1 may have a function independent of FANCD2, and at a level downstream of RAD51 foci formation, in addition to deubiquitinating FANCD2.The USP1 and UAF1 complex interacts with RAD51AP1In order to search for the potential role of USP1 or UAF1 in HR repair, we undertook a proteomic approach to identify potential UAF1 or USP1 interacting proteins that regulate HR repair."
reach
"Focusing on autophagy that makes a critical effect on tumor development, researchers have successively identified and screened various specific small-molecule inhibitors of autophagy-related DUBs, including USP1-UAF1 complex inhibitors GW7647 and Pimozide, which can successfully reverse NSCLC cell resistance to chemotherapeutic drugs (Chen et al., 2011)."
reach
"Indeed, the structure of USP1‐UAF1 complex bound to ID2 ‐DNA revealed ID2 movements towards the open‐state conformation, affecting not only FANCI helices in the region where ubiquitin‐conjugation occurs, but also, and most profoundly, the FANCD2 N‐terminus involved in interaction with FANCI's ubiquitin (Rennie et al, 2021)."
reach
"Given the structural similarity between these USP14 inhibitors and the USP1 inhibitor ML323 (12), it is tempting to speculate that this equally drug-like lead may bind into a similar region of the USP1/UAF1 complex.In June 2015, Genentech and Almac Discovery announced a collaboration on an undisclosed USP target, whereby Almac Discovery's small-molecule leads would provide the starting point for a two-year joint research programme."
sparser
"A recent report has showed that the di-monoubiquitinated frog ID2 complex (both FANCD2 and FANCI monoubiquitinated), generated in the presence of plasmid DNA, remains resistant to deubiquitination by a GST-tagged USP1–UAF1 complex and only the removal of DNA permits efficient DUB activity ( xref )."
sparser
"Interestingly, and despite the presence of FANCI, which may also recruit the DUB complex ( xref ), USP1 ΔN –UAF1 and USP1 R22E –UAF1 are not able to fully deubiquitinate hs FANCD2-Ub, suggesting that any indirect recruitment of USP1–UAF1 by FANCI does not completely compensate for the loss of the USP1 N-terminus."
sparser
"Yu et al reported that the USP1‐WDR48 complex stabilized TANK‐binding kinase 1 by moving its K48‐linked polyubiquitination, and this process could be attenuated using the inhibitor ML‐323. xref Intriguingly, we found that WDR48 was also overexpressed in HCC tissues compared with normal tissues and was associated with poor prognosis in patients."
sparser
"In the past years, several agents aimed at USP1 have been reported, such as pimozide and ML‐323. xref , xref These agents inhibit the activity of USP1‐WDR48 complex in a non‐competitive manner. xref In the present study, we demonstrated that siRNA‐USP1 transfection or ML‐323 treatment decreased the proliferation of HCC cells."
reach
"The inhibition of the deubiquitinase complex USP1-UAF1 or the expression of constitutive fusion of ubiquitin to PCNA provides the missing clue required for DNA polymerase η recruitment and thereafter the introduction of A/T base pair (bp) mutations during the process of IgV gene diversification."
reach
"The cata-lytic activity and stability of USP1 is promoted by itsstoichiometric binding partner UAF1 (USP1 Associated Factor 1; WDR48), a WD40 repeat containing protein.11 Both USP1and UAF1 are regulators of the HR repair, as knockouts ofUSP1 or UAF1 in DT40 cells show reduced HR repair efficiency.12 The USP1 and UAF1 complex also deubiquitinates FANCI, which interacts with FANCD2,13 and a replicative polymerase processive factor PCNA.14 Altogether, USP1 andUAF1 are important contributors to the genome integrity atleast in part by regulating the HR and TLS DNA repairpathways.CONTACT Younghoon Kee Ykee@usf.edu Department of Cell Biology, Microbiology, and Molecular Biology, College of Arts and Sciences, University of South Florida, Tampa, FL 33620, USA."
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"15, NO. 19, 2636-2646 http://dx.doi.org/10.1080/15384101.2016.1209613REPORTThe USP1 and UAF1 complex interacts with RAD51AP1 to promote homologous recombination repairScott Cukrasa, Euiho Leea, Emily Palumboa, Pamela Benavideza, George-Lucian Moldovanb, and Younghoon KeeaaDepartment of Cell Biology, Microbiology, and Molecular Biology, College of Arts and Sciences, University of South Florida, Tampa, FL, USA; bDepartment of Biochemistry and Molecular Biology, Pennsylvania State University College of Medicine, Hershey, PA, USAABSTRACTUSP1 deubiquitinating enzyme and its stoichiometric binding partner UAF1 play an essential role in promoting DNA homologous recombination (HR) repair in response to various types of DNA damaging agents."
sparser
"Recent studies have progressively elucidated the immunoregulatory roles of the USP1‐UAF1 complex. [ xref ] For instance, one study indicates that USP1‐UAF1 modulates the differentiation of T helper cell 17 and Treg cells by stabilizing Tafazzin, [ xref ] while another demonstrates that USP1‐UAF1 stabilizes NLR family pyrin domain containing 3 (NLRP3), thereby activating the NLRP3‐mediated inflammatory phenotype. [ xref ] In the context of type I interferon pathway regulation during RNA virus infection, a report suggests that USP1‐UAF1 augments antiviral responses by stabilizing TBK1. [ xref ] However, the regulatory mechanism of USP1‐UAF1 within the STING‐TBK1‐IRF3 signaling pathway remains uncharted."
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"Nonetheless, both studies revealed a critical role for the UAF1-RAD51AP1 interaction in promoting HR repair, through the DYDL sequence.In summary, this study : 1) dissected the new function of the USP1 and UAF1 complex in HR repair, 2) presents a new mode of regulation of HR repair proteins, and 3) supports the model that UAF1 is a substrate adaptation molecule for USP1."
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"Indeed, RAD51AP1 was identified as a ubiquitinated protein in a proteomic study.40 However, we were unable to detect the ubiquitinated species of RAD51AP1, thus we are not able to conclude whether the USP1 and UAF1 complex protects RAD51AP1 from proteasomal degradation by deubiquitination.RAD51AP1 promotes the HR repair by promoting the recombinase activities of RAD51 and its meiotic counterpart DMC1.4,5,19-21 Unlike the RAD51 paralogs (e.g. RAD51C), depletion of RAD51AP1 does not affect the foci formation of RAD51.4,5 Thus it was proposed that RAD51AP1 acts at a later step of the RAD51-ssDNA nucleofilament formation."
sparser
"GSK1904529 (IGF1R inhibitor), Lapatinib (EGFR inhibitor), Linsitinib (IGF1R inhibitor), ML323 (selective USP1–UAF1 inhibitor), Dihydrorotenone (mitochondrial inhibitor), OSI − 027 (mTOR inhibitor), TAF1_5496 (TAF1 inhibitor), Tamoxifen (selective estrogen response regulator), Vorinostat (Histone deacetylases inhibitor), Gallibiscoquinazole, OF − 1-1853, Sinularin (natural product from soft coral) were more resistant in high-risk group in GSE39582 and GSE17536 cohorts (Fig. xref C, xref )."
reach
"Whether the UAF1 binding to USP1 is also involved in Snail de-ubiquitination and whether USP1 phosphorylation on S42 and/or S331 affects USP1/UAF interaction have not yet been clarified and certainly need to be addressed in future studies.Recent studies suggest that the cancer stem cell status is not static but dynamic, that the differentiation into a cancer stem cell is not a monodirectional process, and that bulk cancer cells can also be reprogrammed into stem cells, exploiting plasticity programs (35)."
reach
"ATR-mediated phosphorylation of the USP1-UAF1 complex and of its substrate, ubiquitylated FANCI-D2, has also been shown to regulate the formation of the enzyme-substrate complex for efficient Ub removal.Related to USP1-UAF1, is how some DUBs function in multiple different DDR signalling pathways."
sparser
"We verified the association of UAF1 and USP1 in testes by co-IP and immunoblotting, confirming (1) that USP1 was present among the immunoprecipitated proteins from wild-type (WT) testes using an anti-UAF1 antibody and (2) that UAF1 was present among the immunoprecipitated proteins from WT testes using an anti-USP1 antibody ( xref C and 1D)."
sparser
"These drugs included various small molecule inhibitors, including MEK/ERK pathway inhibitors (selumetinib, trametinib, and VX‐11e), a survivin inhibitor (sepantronium bromide), a CDK1 inhibitor (RO‐3306), a BCR‐ABL inhibitor (nilotinib), a USP1‐UAF1 inhibitor (ML 323), an estrogen receptor antagonist (fulvestrant), a PORCN/Wnt inhibitor (Wnt‐C59), an IDH inhibitor (AGI‐6780), and cisplatin sensitivity (oxaliplatin; Figure xref )."
reach
"Positive regulators like neuregulin receptor degradation protein 1 (Nrdp1), RING finger protein 128 (RNF128), tyrosine kinase Src, Raf kinase inhibitory protein (RKIP), Glycogen Synthase Kinase-3 Beta (GSK3β), DNA methyltransferases 3A (Dnmt3a), histone deacetylase 3 (HDAC3), and tripartite motif protein 9 (TRIM9) could enhance the autophosphorylation or activation, and USP1-UAF1 complex could stabilizes TBK1 to enhance its function."
sparser
"Notably, ID1 is a substrate of USP1 [ xref ], and blockade of USP1 by ML323 (a selective inhibitor of the USP1-UAF1 complex) not only promoted CD8 + T-cell infiltration and function but also markedly inhibited the liver metastasis of CRC in an orthotopic liver metastatic syngeneic mouse model [ xref ] ( Table xref ) ."
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"Last year, great strides were made in understanding the functional significance of ELG1 when it was discovered that the human homolog of ELG1 (which is also called ATAD5) not only interacts with PCNA and colocalizes with the sliding clamp at stalled replication forks with distinct foci structures in the nucleus, but also associates with the USP1 and UAF1 complex."
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"For high-throughput screening, we used the recombinant USP1 and UAF1 complex in a 4 muL assay volume to screen several libraries of 9,525 bioactive small molecules in qHTS mode, by testing each compound as a dilution series spanning the concentration range of 0.46 muM to 114 muM (see Experimental Procedures)."
sparser
"Although we cannot exclude the possibility of a UAF1‐ssDNA binding event promoting ID2 deubiquitination, there is no evidence in support of dsDNA‐UAF1 binding influencing ID2 deubiquitination: cryo‐EM analysis of in vitro assembled USP1‐UAF1‐ID2 Ub ‐DNA complexes, suggests that formation of a ternary complex is favoured, with both dsDNA and USP1‐UAF1 preferentially binding ubiquitinated ID2, rather than each other (Rennie et al , xref )."
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"In cluster 1, one of the highly ranked complexes searched against CORUM is the USP1 and UAF1 complex which was reported as a schizophrenic candidate complex, and its inhibitor, Pimozide, is one of the antipsychotics with the neuroleptic property of treating schizophrenic patients [XREF_BIBR]."
sparser
"19, 2636–2646 http://dx.doi.org/10.1080/15384101.2016.1209613
REPORT
The USP1-UAF1 complex interacts with RAD51AP1 to promote homologous recombination repair
Scott Cukrasa, Euiho Leea, Emily Palumboa, Pamela Benavideza, George-Lucian Moldovanb, and Younghoon Keea
aDepartment of Cell Biology, Microbiology, and Molecular Biology, College of Arts and Sciences, University of South Florida, Tampa, FL, USA; bDepartment of Biochemistry and Molecular Biology, Pennsylvania State University College of Medicine, Hershey, PA, USA
ABSTRACT
USP1 deubiquitinating enzyme and its stoichiometric binding partner UAF1 play an essential role in promoting DNA homologous recombination (HR) repair in response to various types of DNA damaging agents."
reach
"Setting up and exploring an Arg and N-end rule based in vitro ubiquitylation and degradation system, with the Usp1 and Uaf1 complex as its autocleaved substrate, is likely to be a particularly informative approach to mechanistic questions in this arena, and to some physiological questions as well."
sparser
"In particular, phosphorylation of FANCI provides an on/off switch for the FA pathway activation, as it is required for the monoubiquitination of both FANCI and FANCD2 and also provides a protective mechanism toward the action of the deubiquitinating enzyme USP1:UAF1 ( xref , xref , xref )."
sparser
"USP1
associates with UAF1 (USP1-associated factor 1), resulting in the
heterodimeric USP1/UAF1 complex that is required for deubiquitinase
activity. xref The USP1/UAF1 complex has been
shown to regulate the tolerance of DNA damage induced by DNA cross-linking
agents through deubiquitination of PCNA (proliferating cell nuclear
antigen) xref and FANCD2 (Fanconi anemia complementation
group D2), xref which are proteins that function
in translesion synthesis and the Fanconi anemia pathway, respectively."
sparser
"It was shown that efficient mono-Ub–PCNA deubiquitination by UAF1–USP1 requires an additional factor, ATPase family AAA+ domain-containing protein 5 (ATAD5), the largest subunit of the PCNA-unloading complex ( xref ), while UAF1–USP1 deubiquitinates other substrates such as FANCD2–FANCI on its own ( xref xref – xref )."
sparser
"Focusing on autophagy that makes a critical effect on tumor development, researchers have successively identified and screened various specific small-molecule inhibitors of autophagy-related DUBs, including USP1-UAF1 complex inhibitors GW7647 and Pimozide, which can successfully reverse NSCLC cell resistance to chemotherapeutic drugs ( xref )."
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"Cell CycleISSN : 1538-4101 (Print) 1551-4005 (Online) Journal homepage : http://www.tandfonline.com/loi/kccy20The USP1 and UAF1 complex interacts with RAD51AP1 to promote homologous recombination repairScott Cukras, Euiho Lee, Emily Palumbo, Pamela Benavidez, George-Lucian Moldovan & Younghoon KeeTo cite this article : Scott Cukras, Euiho Lee, Emily Palumbo, Pamela Benavidez, GeorgeLucian Moldovan & Younghoon Kee (2016) The USP1 and UAF1 complex interacts with RAD51AP1 to promote homologous recombination repair, Cell Cycle, 15:19, 2636-2646, DOI : 10.1080/15384101.2016.1209613 To link to this article : https://doi.org/10.1080/15384101.2016.1209613View supplementary materialAccepted author version posted online : 27 Jul 2016."
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"A recent report has showed that the di-monoubiquitinated frog ID2 complex (both FANCD2 and FANCI monoubiquitinated), generated in the presence of plasmid DNA, remains resistant to deubiquitination by a GST-tagged USP1–UAF1 complex and only the removal of DNA permits efficient DUB activity (32)."
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"In this study, a high-throughput screening of nearly 10,000 bioactive small-molecules, followed by further validation and characterization of the best candidates, led to the identification of two potent and highly selective reversible inhibitors of the enzymatic activity of the USP1 and UAF1 complex, pimozide and GW7647."
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"Thus, we propose that the interaction with RAD51AP1 at the post-RAD51 foci formation step is an additional mechanism that regulates the HR repair by the USP1 and UAF1 complex, independently of FANCD2 deubiquitination.Our analysis showed that RAD51AP1 is a mildly unstable protein, and depletion of USP1 or UAF1 greatly decreases the stability."
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"Additionally, the same study disrupted the NHEJ pathway, via knockout of Ku70 (also known as XRCC6 or X-Ray repair cross complementing 6), in UAF1 -/-/- cells which restored the cellular resistance to camptothecin and the PARP inhibitor suggesting that the USP1 and UAF1 complex promotes HR, at least in part by suppressing NHEJ."
sparser
"To further confirm this, we irradiated cells with UV to induce RAD18-mediated PCNA mono-ubiquitination [ xref – xref ] or treated a ubiquitin-specific protease 1 (USP1) inhibitor (hereafter USP1i, ML323) to block Ub-PCNA deubiquitination by the USP1-USP1-associated factor 1 (UAF1) complex [ xref ]."
sparser
"The WDR48–USP1 complex acts as a regulator during DNA damage, especially in translation synthesis and Fanconi anaemia pathway. xref , xref , xref Recent studies have found that the USP1 complex can downregulate the polyubiquitination of TAK1 and mediate its stability in vitro. xref However, few reports exist on the role and mechanism of WDR48 in HCC."
sparser
"One of the best characterized human DUBs is
ubiquitin-specific protease 1 (USP1), which plays an important role
in the cellular response to DNA damage. xref Indeed, the complex of USP1 with USP1-associated factor 1 (UAF1)
acts in two important DDR pathways, namely, translesion synthesis
(TLS) and interstrand cross-links (ICLs), through the deubiquitination
of proliferating cell nuclear antigen (PCNA) and Fanconi anemia complementation
group 2D (FANCD2)."
sparser
"These compounds demonstrate the possibility
of inhibiting the USP1-UAF1 complex with small molecules and synergize
with cisplatin in cisplatin-resistant cells. xref However, both compounds are known to bind several DUBs, deSUMOylase,
and cysteine proteases, which has limited their utility as USP1-UAF1
chemical probes."
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"Although the only BRCA2 mutant cell line in this dataset, PEO1, was not sensitive to USP1 depletion, presented but unpublished work from us and others have shown USP1 inhibitor activity in BRCA2 mutant PDX models.To evaluate USP1 as a potential drug target, we synthesized previously reported inhibitors of the USP1–UAF1 complex, ML323 and I-138 (Fig. 1C; refs."
sparser
"Contrary to this finding, we find that inhibition of USP1 with C527 increased TB40/E genome replication in CD34+ HPCs, suggesting that at least the UAF1-USP1 complex is suppressive to viral genome replication, consistent with the role of UL138 in suppressing replication [ xref ]."
sparser
"In cells, unloading of FANCD2 Ub –FANCI Ub may be instigated by DVC1-p97 [ xref ] but removal of DNA from the FANCD2 Ub –FANCI Ub complex in vitro is sufficient to again permit the activity of USP1:UAF1 on the complex [ xref , xref ] highlighting the critical role that DNA plays in stabilizing the structure of the heterodimer."
reach
"Notably, ID1 is a substrate of USP1 [51], and blockade of USP1 by ML323 (a selective inhibitor of the USP1-UAF1 complex) not only promoted CD8 T-cell infiltration and function but also markedly inhibited the liver metastasis of CRC in an orthotopic liver metastatic syngeneic mouse model [19] (Table 1)."
sparser
"Interestingly, whereas, as expected xref , xref , xref , UAF1 deficient cells showed an elevated level of monoubiquitinated FANCD2, but cells expressing either the UAF1 3A or UAF1 11A mutant harbored mostly deubiquitinated FANCD2 (Fig. xref ), indicating that the UAF1 mutations have little or no impact on the FANCD2-specific DUB activity of the USP1-UAF1 complex in cells."
sparser
"Several findings have emerged, namely: (1) occurrence of USP1-UAF1 mediated-deubiquitination of FANCD2 on DNA; (2) identification of UAF1 amino acid residues important for DNA binding activity; (3) demonstration that UAF1 DNA binding as being required for FANCD2 deubiquitination within the context of the USP1-UAF1 complex; (4) documentation that DNA binding by RAD51AP1 can substitute for that of UAF1 in FANCD2 deubiquitination within the context of the trimeric USP1-UAF1-RAD51AP1 complex; and (5) revelation that UAF1 and RAD51AP1 provide redundant functions for FANCD2 deubiquitination in cells, thus uncovering a role of RAD51AP1 in the FA DNA damage response in addition to its well-documented DNA repair function."
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"10 The USP1 and UAF1 complex has been shown to regulate the tolerance of DNA damage induced by DNA cross linking agents through deubiquitination of PCNA (proliferating cell nuclear antigen) 11 and FANCD2 (Fanconi anemia complementation group D2), 12 which are proteins that function in translesion synthesis and the Fanconi anemia pathway, respectively."
sparser
"Given that human FANCI and FANCD2 share relatively low sequence identity with their Xenopus counterparts (~65% for FANCI and ~55% for FANCD2), it seems plausible that the requirement of DNA for efficient FANCD2 deubiquitination by human USP1-UAF1 becomes relaxed with the Xenopus protein."
sparser
"Furthermore, BRCA1-deficient cells expressing a truncated variant of USP1 deficient in DNA binding fail to deliver USP1-UAF1 to replication forks to act on the ubiquitinated form of the proliferating cell nuclear antigen (PCNA) and are, consequently, impaired for the ability to stabilize stressed DNA replication forks xref ."
sparser
"Proteins involved in the formation of large protein complexes were also selected, such as UAF1, which is part of the deubiquitinating enzyme heterodimeric complex USP1-UAF1 [ xref ]; The Cyclin-Dependent Kinase 7 (CDK7), a subunit of the transcription/DNA repair factor TFIIH [ xref ], is both an effector kinase which phosphorylates both RNA Polymerase II and other transcription factors and a Cdk Activating Kinase (CAK) for essential CDKs [ xref ]; VPS26A, which is a subunit of the core ‘Retromer’ complex that mediates endosomal protein sorting and trafficking [ xref ]; and Munc18a and Munc18c, which regulate SNARE-complexes that mediate vesicle fusion [ xref ]."
sparser
"The experimental systems that we and others xref , xref have developed will be useful for answering mechanistic questions regarding the intracellular functions of the USP1-UAF1 and for establishing the biological impact of cancer-associated mutations in the USP1-UAF1-RAD51AP1 complex."
sparser
"The inhibition of the deubiquitinase complex USP1-UAF1 or the expression of constitutive fusion of ubiquitin to PCNA provides the missing clue required for DNA polymerase η recruitment and thereafter the introduction of A/T base pair (bp) mutations during the process of IgV gene diversification."
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"These data altogether suggest that the USP1 and UAF1 complex promotes HR repair via multiple mechanisms : through FANCD2 deubiquitination, as well as by interacting with RAD51AP1.ARTICLE HISTORY Received 15 April 2016 Revised 14 June 2016 Accepted 29 June 2016KEYWORDS deubiquitinating enzyme; homologous recombination repair; RAD51AP1; UAF1; USP1IntroductionDNA double strand breaks (DSBs) are highly lethal LESIONS that must be repaired before cell division ensues."
sparser
"To determine if the UAF1-USP1 interaction was involved in UL138-mediated regulation of pSTAT1, we inhibited UAF1-USP1 interactions with the chemical inhibitor, C527 [ xref ] at a low concentration of 0.88 μ M which specifically inhibits only UAF1-USP1 deubiquitinating activity [ xref ], in fibroblasts infected with WT or Δ UL138 STOP viruses ( xref )."
sparser
"Since USP1‐UAF1 is a specific deubiquitinase (DUB) for FANCD2 xref , xref , we wanted to assess whether alternative DUBs, that lack FANCD2 specificity (i.e. target structurally diverse substrates), are also able to access the ubiquitin on FANCD2 when this is in complex with I or I Ub ."
sparser
"Positive regulators like neuregulin receptor degradation protein 1 (Nrdp1), RING finger protein 128 (RNF128), tyrosine kinase Src, Raf kinase inhibitory protein (RKIP), Glycogen Synthase Kinase-3 Beta (GSK3β), DNA methyltransferases 3A (Dnmt3a), histone deacetylase 3 (HDAC3), and tripartite motif protein 9 (TRIM9) could enhance the autophosphorylation or activation, and USP1-UAF1 complex could stabilizes TBK1 to enhance its function."
reach
"The USP1 and UAF1 complex is emerging as a novel target for cancer treatment, and inhibitors of USP1 catalytic activity have been reported to reverse the resistance to platinum based chemotherapeutic drugs in NSCLC cells [XREF_BIBR, XREF_BIBR] and to inhibit the growth of leukemic cell lines [XREF_BIBR]."
sparser
"In HPV infection, viral helicase, E1, promotes UAF1-USP1 activity for viral genome replication [ xref , xref ] and the high-risk HPV oncogenes, E6 and E7, delay UAF1-USP1 mediated deubiquitination of the Fanconi anemia (FA) DNA crosslink repair, contributing to genomic instability [ xref ]."
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"Previous reports show that USP1 and UAF1 form a complex and function together, and UAF1 significantly enhances USP1 activity by stabilizing its protein levels and mediating its access to substrates.14 20 USP1 and UAF1 form a complex in the cytoplasm that is subsequently translocated to the nucleus through active nuclear import mediated by the two USP1 NLSs."
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"USP1 is stabilized and activated by UAF1 in vivo and in vitro.14 Previous studies have shown that USP1 and UAF1 form a complex in the cytoplasm that is subsequently translocated to the nucleus through active nuclear import mediated by the two USP1 NLSs.30 Conditional knockout of Uaf1 in the testes results in reduced activity and impaired nuclear-cytoplasmic translocation in round spermatids, thus compromising their function."
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"The USP1 and UAF1 complex is also recruited to mono-ubiquitylated PCNA via an interaction between UAF1 and the protein ELG1, a protein involved in replication complexes and in loading PCNA onto DNA for efficient replication, and independently identified as a factor required to suppress genomic instability [XREF_BIBR, XREF_BIBR]."
sparser
"Whereas the FANCI interaction with the ubiquitin conjugated to FANCD2 has no protective role against USP1‐UAF1 mediated FANCD2 deubiquitination, the interaction of FANCI's ubiquitin with FANCD2 (in I Ub D2‐DNA and I Ub D2 Ub ‐DNA complexes) efficiently protects against FANCI deubiquitination."
sparser
"To test whether this extended interface is involved in protecting FANCI from deubiquitination, we mutated H209, V243 and P244 of FANCD2 to alanine residues, and assessed I Ub ‐DNA deubiquitination by USP1‐UAF1 in the presence of wild‐type (D2 WT ) or H209A/V243A/P244A mutated (D2 H209A,VP243AA ) FANCD2."
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"Among these ten genes, the high expression of seven genes was significantly related to poor OS (BPTF, SETD2, SMARCC1, UBXN7, SMC3, PBRM1 and SF3B1) (Figure 9C), while the other three genes showed no significance (ATRX, SIN3A and USP34) (Figure S3).3.8
Validation of the interaction between USP1 and WDR48 in HCC cell lines."
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"In addition, we confirmed the interaction between USP1 and WDR48 in HCC cell lines using co‐immunoprecipitation, and the results showed that USP1 interacted with WDR48 in MHCC97H and SK‐Hep‐1 cells (Figure 10H).3.9
SiRNA-USP1 transfection or ML-323 treatment decreased the proliferation of HCC cells."
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"142 Liang et al. 143 investigated the binding effects of inhibitor ML323 on the human deubiquitinase complex USP1 and UAF1 using HDX MS and showed four moderately protected and two strongly protected peptides located outside the USP1 catalytic site, supporting the idea that ML323 is an allosteric inhibitor of USP1-UAF1."
sparser
"The effect of FANCI's ubiquitination may be on rendering the doubly ubiquitinated ID2 complex unamenable to USP1‐UAF1 modulation, for example by stabilizing an ID2 conformational change induced by FANCD2 ubiquitination or inducing further minor conformational changes that stabilize the complex."
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"As RAD51AP1 amplification is associated with multiple types of cancers,18,43-45 therapeutic targeting of the USP1 and UAF1 complex may be beneficial in suppressing these tumors.Materials and methodsCell lines, plasmids and chemical reagentsHeLa, 293T, and U2OS cells were cultured in Dulbecco 's Modified Eagle 's Medium (DMEM), and HEY cells (a gift from Dr. Meera Nanjundan) were grown in RPMI media."
sparser
"Lastly, FANCD2 deubiquitination, which also involves a conformational transition step, whereby ID2 Ub ‐DNA complex opens‐up a bit upon USP1‐UAF1 binding (Rennie et al , xref ), also progresses much faster when the initial closed ID2 Ub ‐DNA state is compromised into a more open‐state via the FANCI R1285A mutation (Wang et al , xref )."