Kim, Sung Ah
(Disease Target Structure Research Center, Korea Research Institute of Bioscience and Biotechnology)
,
Jo, Seung-Hyun
(Disease Target Structure Research Center, Korea Research Institute of Bioscience and Biotechnology)
,
Cho, Jin Hwa
(Disease Target Structure Research Center, Korea Research Institute of Bioscience and Biotechnology)
,
Yu, Min Yeong
(Disease Target Structure Research Center, Korea Research Institute of Bioscience and Biotechnology)
,
Shin, Ho-Chul
(Disease Target Structure Research Center, Korea Research Institute of Bioscience and Biotechnology)
,
Kim, Jung-Ae
(Personalized Genomic Medicine Research Center, Korea Research Institute of Bioscience and Biotechnology)
,
Park, Sung Goo
(Disease Target Structure Research Center, Korea Research Institute of Bioscience and Biotechnology)
,
Park, Byoung Chul
(Disease Target Structure Research Center, Korea Research Institute of Bioscience and Biotechnology)
,
Kim, Sunhong
(Disease Target Structure Research Center, Korea Research Institute of Bioscience and Biotechnology)
,
Kim, Jeong-Hoon
(Disease Target Structure Research Center, Korea Research Institute of Bioscience and B)
Aryl hydrocarbon receptor nuclear translocator (ARNT) plays an essential role in maintaining cellular homeostasis in response to environmental stress. Under conditions of hypoxia or xenobiotic exposure, ARNT regulates the subset of genes involved in adaptive responses, by forming heterodimers with h...
Aryl hydrocarbon receptor nuclear translocator (ARNT) plays an essential role in maintaining cellular homeostasis in response to environmental stress. Under conditions of hypoxia or xenobiotic exposure, ARNT regulates the subset of genes involved in adaptive responses, by forming heterodimers with hypoxia-inducible transcription factors (HIF1α and HIF2α) or aryl hydrocarbon receptor (AhR). Here, we have shown that ARNT interacts with DDB1 and CUL4-associated factor 15 (DCAF15), and the aryl sulfonamides, indisulam and E7820, induce its proteasomal degradation through Cullin-RING finger ligase 4 containing DCAF15 (CRL4DCAF15) E3 ligase. Moreover, the two known neo-substrates of aryl sulfonamide, RNA-binding motif protein 39 (RBM39) and RNA-binding motif protein 23 (RBM23), are not required for ARNT degradation. In line with this finding, aryl sulfonamides inhibited the transcriptional activities of HIFs and AhR associated with ARNT. Our results collectively support novel regulatory roles of aryl sulfonamides in both hypoxic and xenobiotic responses.
Aryl hydrocarbon receptor nuclear translocator (ARNT) plays an essential role in maintaining cellular homeostasis in response to environmental stress. Under conditions of hypoxia or xenobiotic exposure, ARNT regulates the subset of genes involved in adaptive responses, by forming heterodimers with hypoxia-inducible transcription factors (HIF1α and HIF2α) or aryl hydrocarbon receptor (AhR). Here, we have shown that ARNT interacts with DDB1 and CUL4-associated factor 15 (DCAF15), and the aryl sulfonamides, indisulam and E7820, induce its proteasomal degradation through Cullin-RING finger ligase 4 containing DCAF15 (CRL4DCAF15) E3 ligase. Moreover, the two known neo-substrates of aryl sulfonamide, RNA-binding motif protein 39 (RBM39) and RNA-binding motif protein 23 (RBM23), are not required for ARNT degradation. In line with this finding, aryl sulfonamides inhibited the transcriptional activities of HIFs and AhR associated with ARNT. Our results collectively support novel regulatory roles of aryl sulfonamides in both hypoxic and xenobiotic responses.
* AI 자동 식별 결과로 적합하지 않은 문장이 있을 수 있으니, 이용에 유의하시기 바랍니다.
제안 방법
, 2017). The stable interactions observed between ARNT and subunits of CRL4DCAF15 led us to investigate whether aryl sulfonamides affect the stability of ARNT. To this end, levels of ARNT and RBM39 were examined in the presence of indisulam and E7820 via western blot.
3B). To ascertain whether the indisulam-induced decrease in ARNT was dependent on proteasome and Cullin-containing E3 ligases, we examined the effects of bortezomib, a proteasome inhibitor, and MNL4924, a Neddylation inhibitor, along with indisulam. Interestingly, co-treatment with indisulam and bortezomib or MNL4924 prevented degradation of ARNT and RBM39 (Fig.
Binding of aryl sulfonamide with DCAF15 could induce a tilt at the binding interface of ARNT and DCAF15, thereby repositioning ARNT for ubiquitination by the CRL4DCAF15 complex. Further structural analyses, such as crystallization or cryogenic electron microscopy (cryo-EM), of the ARNT-DCAF15-aryl sulfonamide complex are required to test this hypothesis.
The following primers were employed for real-time PCR: DCAF15 (5’-GAGGTCTGCCCAGAAACCAA-3’ and 5’-CTCAGTCAGGTCGCCTACA-C-3'), GAPDH (5’-GGTATCGTGGAAGGACTCATGA-C-3' and 5’-ATGCCAGTGAGCTTCCCGTTCAGC-3'), GLUT1 (5’-AACTCTTCAGCC-AGGGACCT-3' and 5’-CACAGTGAAGATGATGAAGA-3'), LDHA (5’-TTGGTCCAGC-GTAACGTGAA-3' and 5’-CCAGGATGTGTAGCCTTTGA-3'), VEGF (5’-CTACCTCCA-CCATGCCAAGT-3' and 5’-GCAGTAGCTGCGCTCATAGA-3'), TGFα (5’-CTGCCCG-CCCGCCCGTAAAA-3' and 5’-C-CGCATGCTCACAGCGTGCA-3'), and β-actin (5’-C-ATGTACGTTGCTATCCAGGC-3' and 5’-CTCCTTAATGTCACGCACGAT-3').
대상 데이터
The human gene encoding full-length DCAF15 was purchased from Novoprolabs in China (703683-1, NM_138353). Full-length and truncated DCAF15 were subcloned into pcDNA3.
The reagents used included MLN4924 (505477; Calbiochem, USA), Bortezomib (S1013; Selleckchem, USA), Indisulam (SML1225; Sigma-Aldrich, USA), and E7820 (25502; Cayman Chemical Company, USA).
이론/모형
We thus examined whether the function of ARNT is affected by indisulam. Transcriptional activity of AhR/ARNT was examined using the XRE reporter luciferase assay. Treatment with a known xenobiotic, 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD), induced XRE reporter activity > 100-fold in HepG2 cells, which was suppressed by indisulam (Fig.
참고문헌 (39)
Abel J. Haarmann-Stemmann T. 2010 An introduction to the molecular basics of aryl hydrocarbon receptor biology Biol. Chem. 391 1235 1248 10.1515/bc.2010.128 20868221
Assi R. Kantarjian H.M. Kadia T.M. Pemmaraju N. Jabbour E. Jain N. Daver N. Estrov Z. Uehara T. Owa T. 2018 Final results of a phase 2, open-label study of indisulam, idarubicin, and cytarabine in patients with relapsed or refractory acute myeloid leukemia and high-risk myelodysplastic syndrome Cancer 124 2758 2765 10.1002/cncr.31398 29660836
Bussiere D.E. Xie L. Srinivas H. Shu W. Burke A. Be C. Zhao J. Godbole A. King D. Karki R.G. 2020 Structural basis of indisulam-mediated RBM39 recruitment to DCAF15 E3 ligase complex Nat. Chem. Biol. 16 15 23 10.1038/s41589-019-0411-6 31819272
Du X. Volkov O.A. Czerwinski R.M. Tan H. Huerta C. Morton E.R. Rizzi J.P. Wehn P.M. Xu R. Nijhawan D. 2019 Structural basis and kinetic pathway of RBM39 recruitment to DCAF15 by a sulfonamide molecular glue E7820 Structure 27 1625 1633 e3 10.1016/j.str.2019.10.005 31693911
Gradin K. McGuire J. Wenger R.H. Kvietikova I. Toftgård R. Tora L. Gassmann M. Poellinger L. 1996 Functional interference between hypoxia and dioxin signal transduction pathways: competition for recruitment of the Arnt transcription factor Mol. Cell. Biol. 16 5221 5231 10.1128/MCB.16.10.5221 8816435
Haddad R.I. Weinstein L.J. Wieczorek T.J. Bhattacharya N. Raftopoulos H. Oster M.W. Zhang X. Latham V.M. Costello R. Faucher J. 2004 A phase II clinical and pharmacodynamic study of E7070 in patients with metastatic, recurrent, or refractory squamous cell carcinoma of the head and neck: modulation of retinoblastoma protein phosphorylation by a novel chloroindolyl sulfonamide cell cycle inhibitor Clin. Cancer Res. 10 4680 4687 10.1158/1078-0432.CCR-04-0229 15269140
Han T. Goralski M. Gaskill N. Capota E. Kim J. Ting T.C. Xie Y. Williams N.S. Nijhawan D. 2017 Anticancer sulfonamides target splicing by inducing RBM39 degradation via recruitment to DCAF15 Science 356 eaal3755 10.1126/science.aal3755 28302793
Hannah J. Zhou P. 2015 Distinct and overlapping functions of the cullin E3 ligase scaffolding proteins CUL4A and CUL4B Gene 573 33 45 10.1016/j.gene.2015.08.064 26344709
Hsiehchen D. Goralski M. Kim J. Xie Y. Nijhawan D. 2020 Biomarkers for RBM39 degradation in acute myeloid leukemia Leukemia 34 1924 1928 10.1038/s41375-020-0729-9 32042080
Hughes S.J. Ciulli A. 2017 Molecular recognition of ternary complexes: a new dimension in the structure-guided design of chemical degraders Essays Biochem. 61 505 516 10.1042/EBC20170041 29118097
Huttlin E.L. Bruckner R.J. Paulo J.A. Cannon J.R. Ting L. Baltier K. Colby G. Gebreab F. Gygi M.P. Parzen H. 2017 Architecture of the human interactome defines protein communities and disease networks Nature 545 505 509 10.1038/nature22366 28514442
Huttlin E.L. Ting L. Bruckner R.J. Gebreab F. Gygi M.P. Szpyt J. Tam S. Zarraga G. Colby G. Baltier K. 2015 The BioPlex network: a systematic exploration of the human interactome Cell 162 425 440 10.1016/j.cell.2015.06.043 26186194
Jung D.J. Na S.Y. Na D.S. Lee J.W. 2002 Molecular cloning and characterization of CAPER, a novel coactivator of activating protein-1 and estrogen receptors J. Biol. Chem. 277 1229 1234 10.1074/jbc.M110417200 11704680
Kang Y.K. Putluri N. Maity S. Tsimelzon A. Ilkayeva O. Mo Q. Lonard D. Michailidis G. Sreekumar A. Newgard C.B. 2015 CAPER is vital for energy and redox homeostasis by integrating glucose-induced mitochondrial functions via ERR-alpha-Gabpa and stress-induced adaptive responses via NF-kappaB-cMYC PLoS Genet. 11 e1005116 10.1371/journal.pgen.1005116 25830341
Kobayashi A. Numayama-Tsuruta K. Sogawa K. Fujii-Kuriyama Y. 1997 CBP/p300 functions as a possible transcriptional coactivator of Ah receptor nuclear translocator (Arnt) J. Biochem. 122 703 710 10.1093/oxfordjournals.jbchem.a021812 9399571
Kronke J. Udeshi N.D. Narla A. Grauman P. Hurst S.N. McConkey M. Svinkina T. Heckl D. Comer E. Li X. 2014 Lenalidomide causes selective degradation of IKZF1 and IKZF3 in multiple myeloma cells Science 343 301 305 10.1126/science.1244851 24292625
Mai S. Qu X. Li P. Ma Q. Cao C. Liu X. 2016 Global regulation of alternative RNA splicing by the SR-rich protein RBM39 Biochim. Biophys. Acta 1859 1014 1024 10.1016/j.bbagrm.2016.06.007 27354116
Maltepe E. Schmidt J.V. Baunoch D. Bradfield C.A. Simon M.C. 1997 Abnormal angiogenesis and responses to glucose and oxygen deprivation in mice lacking the protein ARNT Nature 386 403 407 10.1038/386403a0 9121557
Mandl M. Depping R. 2017 ARNT is a potential direct HIF-1 target gene in human Hep3B hepatocellular carcinoma cells Cancer Cell Int. 17 77 10.1186/s12935-017-0446-2 28855849
Owa T. Yoshino H. Okauchi T. Yoshimatsu K. Ozawa Y. Sugi N.H. Nagasu T. Koyanagi N. Kitoh K. 1999 Discovery of novel antitumor sulfonamides targeting G1 phase of the cell cycle J. Med. Chem. 42 3789 3799 10.1021/jm9902638 10508428
Reisz-Porszasz S. Probst M.R. Fukunaga B.N. Hankinson O. 1994 Identification of functional domains of the aryl hydrocarbon receptor nuclear translocator protein (ARNT) Mol. Cell. Biol. 14 6075 6086 10.1128/MCB.14.9.6075 8065341
Rezvani H.R. Ali N. Nissen L.J. Harfouche G. de Verneuil H. Taïeb A. Mazurier F. 2011 HIF-1alpha in epidermis: oxygen sensing, cutaneous angiogenesis, cancer, and non-cancer disorders J. Invest. Dermatol. 131 1793 1805 10.1038/jid.2011.141 21633368
Scheuermann T.H. Yang J. Zhang L. Gardner K.H. Bruick R.K. 2007 Hypoxia-inducible factors Per/ARNT/Sim domains: structure and function Methods Enzymol. 435 3 24 10.1016/S0076-6879(07)35001-5 17998046
Sogawa K. Nakano R. Kobayashi A. Kikuchi Y. Ohe N. Matsushita N. Fujii-Kuriyama Y. 1995 Possible function of Ah receptor nuclear translocator (Arnt) homodimer in transcriptional regulation Proc. Natl. Acad. Sci. U. S. A. 92 1936 1940 10.1073/pnas.92.6.1936 7892203
Ting T.C. Goralski M. Klein K. Wang B. Kim J. Xie Y. Nijhawan D. 2019 Aryl sulfonamides degrade RBM39 and RBM23 by recruitment to CRL4-DCAF15 Cell Rep. 29 1499 1510 e6 10.1016/j.celrep.2019.09.079 31693891
Uehara T. Minoshima Y. Sagane K. Sugi N.H. Mitsuhashi K.O. Yamamoto N. Kamiyama H. Takahashi K. Kotake Y. Uesugi M. 2017 Selective degradation of splicing factor CAPERalpha by anticancer sulfonamides Nat. Chem. Biol. 13 675 680 10.1038/nchembio.2363 28437394
Van Kesteren C. Beijnen J.H. Schellens J.H. 2002 E7070: a novel synthetic sulfonamide targeting the cell cycle progression for the treatment of cancer Anticancer Drugs 13 989 997 10.1097/00001813-200211000-00002 12439332
Vorrink S.U. Domann F.E. 2014 Regulatory crosstalk and interference between the xenobiotic and hypoxia sensing pathways at the AhR-ARNT-HIF1alpha signaling node Chem. Biol. Interact. 218 82 88 10.1016/j.cbi.2014.05.001 24824450
Wang E. Lu S.X. Pastore A. Chen X. Imig J. Chun-Wei Lee S. Hockemeyer K. Ghebrechristos Y.E. Yoshimi A. Inoue D. 2019 Targeting an RNA-binding protein network in acute myeloid leukemia Cancer Cell 35 369 384 e7 10.1016/j.ccell.2019.01.010 30799057
Wolff M. Jelkmann W. Dunst J. Depping R. 2013 The Aryl Hydrocarbon Receptor Nuclear Translocator (ARNT/HIF-1beta) is influenced by hypoxia and hypoxia-mimetics Cell. Physiol. Biochem. 32 849 858 10.1159/000354487 24081025
Wood S.M. Gleadle J.M. Pugh C.W. Hankinson O. Ratcliffe P.J. 1996 The role of the aryl hydrocarbon receptor nuclear translocator (ARNT) in hypoxic induction of gene expression. Studies in ARNT-deficient cells J. Biol. Chem. 271 15117 15123 10.1074/jbc.271.25.15117 8662957
Zhu Y.X. Braggio E. Shi C.X. Kortuem K.M. Bruins L.A. Schmidt J.E. Chang X.B. Langlais P. Luo M. Jedlowski P. 2014 Identification of cereblon-binding proteins and relationship with response and survival after IMiDs in multiple myeloma Blood 124 536 545 10.1182/blood-2014-02-557819 24914135
※ AI-Helper는 부적절한 답변을 할 수 있습니다.