JPOST Repositoryapplication/xmlhttps://storage.jpostdb.org/JPST002274/files/20220215_hata_20211228_kaito_MDS_L_KO.xlsxhttps://storage.jpostdb.org/JPST002274/files/20220215_hata_20211224_kaito_MDS_L_WT.xlsxhttps://storage.jpostdb.org/JPST002274/files/20211228_kaito_MDS_L_KO.rawhttps://storage.jpostdb.org/JPST002274/files/20211224_kaito_MDS_L_WT.rawprimary200OKProteomicsMasaaki OyamaHomo Sapiens (human)https://repository.jpostdb.org/entry/JPST002274The Institute of Medical Science, The University of TokyojPOSTDNA hypomethylating agents (HMAs) are used for the treatment of myeloid malignancies, although their therapeutic effects have been unsatisfactory. Here we show that CRISPR-Cas9 screening reveals that knockout of topoisomerase 1-binding arginine/serine-rich protein (TOPORS), which encodes a ubiquitin/SUMO E3 ligase, augments the efficacy of HMAs on myeloid leukemic cells with little effect on normal hematopoiesis, suggesting that TOPORS is involved in resistance to HMAs. HMAs are incorporated into the DNA and trap DNA methyltransferase-1 (DNMT1) to form DNA-DNMT1 crosslinks, which undergo SUMOylation, followed by proteasomal degradation. Persistent crosslinking is cytotoxic. The TOPORS RING finger domain, which mediates ubiquitination, is responsible for HMA resistance. In TOPORS knockout cells, DNMT1 is stabilized by HMA treatment due to inefficient ubiquitination, resulting in the accumulation of unresolved SUMOylated DNMT1. This indicates that TOPORS ubiquitinates SUMOylated DNMT1, thereby promoting the resolution of DNA-DNMT1 crosslinks. Consistently, the ubiquitination inhibitor, TAK-243, and the SUMOylation inhibitor, TAK-981, show synergistic effects with HMAs through DNMT1 stabilization. Our study provides a novel HMA-based therapeutic strategy that interferes with the resolution of DNA-DNMT1 crosslinks.Inhibition of TOPORS ubiquitin ligase augments the efficacy of DNA hypomethylating agents through DNMT1 stabilization.Kaito Satoshi S, Aoyama Kazumasa K, Oshima Motohiko M, Tsuchiya Akiho A, Miyota Makiko M, Yamashita Masayuki M, Koide Shuhei S, Nakajima-Takagi Yaeko Y, Kozuka-Hata Hiroko H, Oyama Masaaki M, Yogo Takao T, Yabushita Tomohiro T, Ito Ryoji R, Ueno Masaya M, Hirao Atsushi A, Tohyama Kaoru K, Li Chao C, Kawabata Kimihito Cojin KC, Yamaguchi Kiyoshi K, Furukawa Yoichi Y, Kosako Hidetaka H, Yoshimi Akihide A, Goyama Susumu S, Nannya Yasuhito Y, Ogawa Seishi S, Agger Karl K, Helin Kristian K, Yamazaki Satoshi S, Koseki Haruhiko H, Doki Noriko N, Harada Yuka Y, Harada Hironori H, Nishiyama Atsuya A, Nakanishi Makoto M, Iwama Atsushi AProteins, Ubiquitinated.biochemical pathways, Forms, CRISPR Locus, CRISPR Loci, Ghrfr, Therapeutic Effects, Clustered Regularly Interspaced Short Palindromic Repeat, HSN1E, Met-1, CRISPR Spacer, DL-Arginine Acetate, c-2k, Cxxc9, Ubiquitin Protein Ligase, Ligase E3, Monohydrate, DNA methyltransferase MmuI, protein, l(2)SH2 0182, Tumor, Medullary, DDM2, Ubiquitin Ligase, Smt3, RP11-508D10.1, SMT3, 2-Amino-3-hydroxypropionic acid, Medullary Hematopoiesis, Tnfsf5, METI, Sumoylations, CG9063, Smt3-protein conjugation, symptoms, cellular degradation, Arg, protein aggregate, Effect, Arrays, T-BAM, treatment, average, thymus nucleic acid, (S)-2-amino-5-guanidinopentanoic acid, type 5 acid phosphatase, Ubiquitylation, tartrate-resistant acid ATPase, High Mobility Protein 20, DNA topoisomerase 1 beta, Elements, catabolism, (S)-2-Amino-5-guanidinovaleric acid, R, DmSmt3, Arginine, Ubiquitin-related 1, Monohydrate DL-Arginine Acetate, DmSUMO-1, BcDNA:GH03694, C-2k, 2R, anon-EST:Posey240, Ligase, CRISPRs, APF-1, TRAP3, T-cell antigen Gp39, DmelCG15104, Tudor repeat associator with PCTAIRE-2, 2-amino-3-hydroxypropanoic acid, blood cell formation, Therapeutic Use, disease management, Therapies, P53BP3, DNA METHYLTRANSFERASE 1, 3-dihydroxy-4-((2-(3-(trifluoromethylsulfanyl)phenyl)pyrazolo(1, biotransformation, Double-Stranded DNA, Malignancies, TNFSF5, ubiquitination, deoxyribonucleic acids, TP53BPL, DNAn, T5ap, Smt3p-protein conjugation, MGC - 45012, DmelCG9063, RING Finger Domain, Dmsmt3, Tumors, L-Arginin, CD40L, 3S, Therapy, screening, CRISPR Spacer Sequences, CRISPR Clusters, Ubiquitin Protein Ligases, DMT1, Ubiquitin Ligase E3, E3, l(2)04493, Tnfrsf5, Arginine Hydrochloride, E3 Ubiquitin, Cd40l, MTE17.2, arginine, Ligases, Hematopoiesis, CRISPR Arrays, lit, Double-Stranded, CRISPR-Cas, Clusters, uae inhibitor MLN7243, Arginin, CEP52, MTE17.1, hCD40L, (Deoxyribonucleotide)n+m, CRISPR Element, Benign, Screenings, 3-Hydroxyalanine, MTE17_1, MTE17_2, TOPORS, Domains, IMD3, secretion, TNF-related activation protein, desoxyribose nucleic acid, Therapeutic Effect, Ly-62, Ubiq, little, L-Arginine, Ubiquitin, DNA methyltransferase I, ADCADN, Element, 5-a)pyrimidin-7-yl)amino)cyclopentyl)methyl sulfamate, Ubiquitin-Protein, BcDNAGH03694, NKTL, Human Ubiquitin, signs, Benign Neoplasms, AW743063, DECREASED DNA METHYLATION 2, AI326936, PITALRE, Treatments, 3L6, Malignant Neoplasms, Cluster, rich, DNA MTase RnoIP, antagonists and inhibitors, dtopors, TrATPase, ds DNA, Spacers, DNA topoisomerase I alpha, IGM, Ubiquitin Protein Ligase E3, L Serine, CD40LG, Spacer Sequences, l(2)SH0182, DNA, MGO1, L-Serine, MLN7243, DmelCG4494, sumoylation, CRISPR-Cas Loci, cdc2l4, DNS, MTase, (Deoxyribonucleotide)n, TRAP, p50, SUMO-Conjugations, Effects, Ly62, Neoplasms, TAPK, Benign Neoplasm, 2-Amino-5-guanidinovaleric acid, L Isomer, Gene, CRISPR Array, ATP Dependent Proteolysis Factor 1, Ubiquitin carboxyl extension protein 80, haemopoiesis, TOP1ALPHA, protein-containing complex, Malignant, TR-AP, cellular catabolism, Deoxyribonucleic acids, L-(+)-arginine, Human, DNA MTase MmuI, AIM, E3 Ligase, M.MmuI, Deoxyribonucleic Acid, (2S)-2-amino-5-(carbamimidamido)pentanoic acid, Harg, GP39, Mass, Gene Products, Screening, 4R)-2, HMG-20, dSmt3, MGOUN 1, CD40-L, DNA METHYLTRANSFERASE, RP31, CRISPR Spacers, RING Finger Motifs, gp39, CD154, p53BP3/LUN, study, CATC4, GKLP, Malignancy, breakdown of chemical, ligand, p53BP3|LUN, inhibiteur, ((1R, Double Stranded, Deoxyribonucleic acid, methyltransferase 1, DNA MTase HsaI., CRISPR Spacer Sequence, CG15104, TOPOISOMERASE 1, CRISPR Sequences, CRISPR Cluster, Neoplasias, CRISPR, cdk9, CRISPR-Cas Locus, 40S ribosomal protein S27a, PCTAIRE2-binding protein, Use, MGC:45012, inhibidor, dTopors, CTK1, TEIF, Trap, M.RnoIP, Loci, ubiquitin, L-Arg, Dnmt1, HIGM1, (Deoxyribonucleotide)m, protein tagging activity, ctk1, 2-amino-5-(carbamimidamido)pentanoic acid, M.HsaI, C78062, Spacer Sequence, Cancer, DNA MTase HsaI, TAK, inhibitors, cellular breakdown, Sumo, P105, NTKL, CRISPR Sequence, findings, Malignant Neoplasm, RING Finger Motif, degradation, protein complex, 2.1.1.37, PCTAIRE2BP, DNAn+1, Proteins, DNMT, inhibitor, Bp50, 2-amino-5-guanidinopentanoic acid, METHYLTRANSFERASE 2, ACP5, MCMT, L-Isomer, DL Arginine Acetate, SPENCDI, Cell, tak, Sequences, SUMO, Serin, Dnmt1o, Ubiquitin A-52 residue ribosomal protein fusion product 1, native protein, L-Isomer Arginine, CG4494, Sequence, Mass Screenings, Ubiquitin-related 2, DNA methyltransferase HsaI, MommeD2, Protein, CRISPR Elements, Neoplasm, Hydrochloride, METHYLTRANSFERASE I, sumo, L Arginine, Spacer, ds-DNA, ATP-Dependent Proteolysis Factor 1, m.MmuI, TOP1, breakdown of molecule, CXXC9, antagonists, covalent modifier, Uses, TOP1BETA, biodegradation, Dm0342, dsmt3, Ubiquitin-Protein Ligase, RING Finger, DNA METHYLTRANSFERASE 01, DMT01, pitalre, (2S)-2-amino-5-guanidinopentanoic acid, E3 Ubiquitin Ligase, Met1, hematopoiesis, MET2, Cancers, Haematopoiesis, MLN-7243, Locus, CXXC-type zinc finger protein 9, Protein Gene Products, SUMO Conjugation, Gene Proteins, LUN, breakdown of substance, Therapeutic, 60S ribosomal protein L40, CDC2L4, blood cell biosynthesis, Desoxyribonukleinsaeure, Array, AW105885, CRISPR Cas Loci, Dnmt, Treatment, Ubiquitin-related, TRACP, SUMO-Conjugation, Ubiquitin-Protein Ligase E3, NeoplasiaDNS, MTase, (Deoxyribonucleotide)n, HSN1E, 2.1.1.37, Met-1, DNAn+1, DNMT, Cxxc9, DNA methyltransferase MmuI, Synthetases, ATP Dependent Proteolysis Factor 1, Ubiquitin carboxyl extension protein 80, Double-Stranded, MCMT, CEP52, Deoxyribonucleic acids, Human, DNA MTase MmuI, (Deoxyribonucleotide)n+m, AIM, Dnmt1o, Ubiquitin A-52 residue ribosomal protein fusion product 1, M.MmuI, Deoxyribonucleic Acid, Ubiquitin-related 2, DNA methyltransferase HsaI, MommeD2, TOPORS, Synthetase, HMG-20, ATP-Dependent Proteolysis Factor 1, ds-DNA, m.MmuI, desoxyribose nucleic acid, RP31, Ubiq, Ubiquitin, CXXC9, p53BP3/LUN, DNA methyltransferase I, covalent modifier, ADCADN, thymus nucleic acid, High Mobility Protein 20, p53BP3|LUN, Met1, Human Ubiquitin, Ubiquitin-related 1, Double Stranded, Deoxyribonucleic acid, DNA MTase HsaI., CG15104, CXXC-type zinc finger protein 9, Ligase, APF-1, 40S ribosomal protein S27a, LUN, DmelCG15104, dTopors, 60S ribosomal protein L40, DNA MTase RnoIP, ligase, M.RnoIP, dtopors, ubiquitin, ds DNA, AW105885, Desoxyribonukleinsaeure, P53BP3, Dnmt1, Dnmt, Ubiquitin-related, Double-Stranded DNA, (Deoxyribonucleotide)m, TP53BPL, DNA, deoxyribonucleic acids, DNAn, protein tagging activity, M.HsaIMds1-Evi1, HEL2, NCTC Clone 929 Cells, Dmel_CG31829, determination, BOPS, FBN, Strain L Cell, Jbo, Earle's Strain L Cells, cul3, Cullin 3, dCul3, l(2)br34, L-Cell, ECTOL1, pafah, Cell Line., L Cell (Cell Line), Mds1, CG31829, L Cell Line, Ubiquitinated, SF3b155, WMS, L929 Cell, DMcul-3, L Cell, CUL3, L929 Cells, L Cells (Cell Line), L, Cell Line L-Cell, guftagu, L-Cell Line, L929, LIS1, Evi1, Strain L Cells, D630039M04Rik, PAFAHA, LIS2, Evi-1, OCTD, Myelodysplastic Syndromes, MDCR, lis2, L-Cells, gft, mdcr, GPHYSD2, Cul3, myelodysplastic syndrome, SGS, l(2)06430, pafah1b1-b, Cell Line L-Cells, L Cells, Proteins, KIAA1546, Cell Lines, CUL-3, MDS, Mds, Cell, gft/dCul-3, br34, ACMICD, PRP10, L929 Cell Line, L-Cell Lines, chemical analysis, KCIP-1, Strain, L Cell Lines, MFS1, Hsh155, PRPF10, WMS2, mds, l35Cd, DmelCG42616, cul-3, BG:DS07851.2, susceptibility to, NCTC Clone 929 of Strain L Cells, Cullin3, SAP155, MASS, dCul-3, PAFAH, CG11861, platelet-activating factor acetylhydrolase IB subunit alpha, Cell Line, L (Cell Line), CG42616, Strain L, Znfpr1b1, 14-3-3E, Dmel_CG11861, SSKS, Cells, l(2)35Cd, Prdm3, assayfalseMS data on ubiquitinated proteinsMass spectrometric analysis of ubiquitinated proteins in MDS-L cellsFri Mar 08 00:00:00 GMT 2024PXD044347960639198387