MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url&gt;http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it <i>circZNF827</i> nucleates a transcription inhibitory complex to balance neuronal differentiation. <i>circZNF827</i> nucleates a transcription inhibitory complex to balance neuronal differentiation. eLife 2020 imex curation Hollensen AK., Thomsen HS., Lloret-Llinares M., Kamstrup AB., Jensen JM., Luckmann M., Birkmose N., Palmfeldt J., Jensen TH., Hansen TB., Damgaard CK. ckd@mbg.au.dk <ckd@mbg.au.dk>; Only protein-protein interactions imex curation human-lan_5 Homo sapiens neuroblastoma cell line LAN-5 lan_5 Human neuroblastoma cell line LAN-5 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Hollensen AK., Thomsen HS., Lloret-Llinares M., Kamstrup AB., Jensen JM., Luckmann M., Birkmose N., Palmfeldt J., Jensen TH., Hansen TB., Damgaard CK. ckd@mbg.au.dk <ckd@mbg.au.dk>; Accepted 2021-APR-08 AT 12:40 BST AT 12:40 BST by KALPANAP eLife 2020 Only protein-protein interactions imex curation imex curation <i>circZNF827</i> nucleates a transcription inhibitory complex to balance neuronal differentiation. <i>circZNF827</i> nucleates a transcription inhibitory complex to balance neuronal differentiation. eLife 2020 imex curation Hollensen AK., Thomsen HS., Lloret-Llinares M., Kamstrup AB., Jensen JM., Luckmann M., Birkmose N., Palmfeldt J., Jensen TH., Hansen TB., Damgaard CK. ckd@mbg.au.dk <ckd@mbg.au.dk>; Only protein-protein interactions imex curation human-293 flp-in Homo sapiens 293 flp-in 293 flp-in confocal microscopy confocal microscopy immunostaining immunostaining Hollensen AK., Thomsen HS., Lloret-Llinares M., Kamstrup AB., Jensen JM., Luckmann M., Birkmose N., Palmfeldt J., Jensen TH., Hansen TB., Damgaard CK. ckd@mbg.au.dk <ckd@mbg.au.dk>; Accepted 2021-APR-08 AT 12:40 BST AT 12:40 BST by KALPANAP eLife 2020 Only protein-protein interactions imex curation imex curation <i>circZNF827</i> nucleates a transcription inhibitory complex to balance neuronal differentiation. <i>circZNF827</i> nucleates a transcription inhibitory complex to balance neuronal differentiation. eLife 2020 imex curation Hollensen AK., Thomsen HS., Lloret-Llinares M., Kamstrup AB., Jensen JM., Luckmann M., Birkmose N., Palmfeldt J., Jensen TH., Hansen TB., Damgaard CK. ckd@mbg.au.dk <ckd@mbg.au.dk>; Only protein-protein interactions imex curation human-293 flp-in Homo sapiens 293 flp-in 293 flp-in anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Hollensen AK., Thomsen HS., Lloret-Llinares M., Kamstrup AB., Jensen JM., Luckmann M., Birkmose N., Palmfeldt J., Jensen TH., Hansen TB., Damgaard CK. ckd@mbg.au.dk <ckd@mbg.au.dk>; Accepted 2021-APR-08 AT 12:40 BST AT 12:40 BST by KALPANAP eLife 2020 Only protein-protein interactions imex curation imex curation hnrpk_human Heterogeneous nuclear ribonucleoprotein K HNRNPK HNRPK Transformation up-regulated nuclear protein protein protein human Homo sapiens Human METEQPEETFPNTETNGEFGKRPAEDMEEEQAFKRSRNTDEMVELRILLQSKNAGAVIGKGGKNIKALRTDYNASVSVPDSSGPERILSISADIETIGEILKKIIPTLEEGLQLPSPTATSQLPLESDAVECLNYQHYKGSDFDCELRLLIHQSLAGGIIGVKGAKIKELRENTQTTIKLFQECCPHSTDRVVLIGGKPDRVVECIKIILDLISESPIKGRAQPYDPNFYDETYDYGGFTMMFDDRRGRPVGFPMRGRGGFDRMPPGRGGRPMPPSRRDYDDMSPRRGPPPPPPGRGGRGGSRARNLPLPPPPPPRGGDLMAYDRRGRPGDRYDGMVGFSADETWDSAIDTWSPSEWQMAYEPQGGSGYDYSYAGGRGSYGDLGGPIITTQVTIPKDLAGSIIGKGGQRIKQIRHESGASIKIDEPLEGSEDRIITITGTQDQIQNAQYLLQNSVKQYSGKFF 0F70EE169B2A064A p14866-1 Heterogeneous nuclear ribonucleoprotein L HNRNPL HNRPL P/OKcl.14 protein protein human Homo sapiens Human MSRRLLPRAEKRRRRLEQRQQPDEQRRRSGAMVKMAAAGGGGGGGRYYGGGSEGGRAPKRLKTDNAGDQHGGGGGGGGGAGAAGGGGGGENYDDPHKTPASPVVHIRGLIDGVVEADLVEALQEFGPISYVVVMPKKRQALVEFEDVLGACNAVNYAADNQIYIAGHPAFVNYSTSQKISRPGDSDDSRSVNSVLLFTILNPIYSITTDVLYTICNPCGPVQRIVIFRKNGVQAMVEFDSVQSAQRAKASLNGADIYSGCCTLKIEYAKPTRLNVFKNDQDTWDYTNPNLSGQGDPGSNPNKRQRQPPLLGDHPAEYGGPHGGYHSHYHDEGYGPPPPHYEGRRMGPPVGGHRRGPSRYGPQYGHPPPPPPPPEYGPHADSPVLMVYGLDQSKMNCDRVFNVFCLYGNVEKVKFMKSKPGAAMVEMADGYAVDRAITHLNNNFMFGQKLNVCVSKQPAIMPGQSYGLEDGSCSYKDFSESRNNRFSTPEQAAKNRIQHPSNVLHFFNAPLEVTEENFFEICDELGVKRPSSVKVFSGKSERSSSGLLEWESKSDALETLGFLNHYQMKNPNGPYPYTLKLCFSTAQHAS DIP protein P14866-1 original sequence version: 145 hnrpu_human Heterogeneous nuclear ribonucleoprotein U HNRNPU SAFA U21.1 HNRPU p120 pp120 C1orf199 GRIP120 Nuclear p120 ribonucleoprotein Scaffold-attachment factor A protein protein human Homo sapiens Human MSSSPVNVKKLKVSELKEELKKRRLSDKGLKAELMERLQAALDDEEAGGRPAMEPGNGSLDLGGDSAGRSGAGLEQEAAAGGDEEEEEEEEEEEGISALDGDQMELGEENGAAGAADSGPMEEEEAASEDENGDDQGFQEGEDELGDEEEGAGDENGHGEQQPQPPATQQQQPQQQRGAAKEAAGKSSGPTSLFAVTVAPPGARQGQQQAGGKKKAEGGGGGGRPGAPAAGDGKTEQKGGDKKRGVKRPREDHGRGYFEYIEENKYSRAKSPQPPVEEEDEHFDDTVVCLDTYNCDLHFKISRDRLSASSLTMESFAFLWAGGRASYGVSKGKVCFEMKVTEKIPVRHLYTKDIDIHEVRIGWSLTTSGMLLGEEEFSYGYSLKGIKTCNCETEDYGEKFDENDVITCFANFESDEVELSYAKNGQDLGVAFKISKEVLAGRPLFPHVLCHNCAVEFNFGQKEKPYFPIPEEYTFIQNVPLEDRVRGPKGPEEKKDCEVVMMIGLPGAGKTTWVTKHAAENPGKYNILGTNTIMDKMMVAGFKKQMADTGKLNTLLQRAPQCLGKFIEIAARKKRNFILDQTNVSAAAQRRKMCLFAGFQRKAVVVCPKDEDYKQRTQKKAEVEGKDLPEHAVLKMKGNFTLPEVAECFDEITYVELQKEEAQKLLEQYKEESKKALPPEKKQNTGSKKSNKNKSGKNQFNRGGGHRGRGGFNMRGGNFRGGAPGNRGGYNRRGNMPQRGGGGGGSGGIGYPYPRAPVFPGRGSYSNRGNYNRGGMPNRGNYNQNFRGRGNNRGYKNQSQGYNQWQQGQFWGQKPWSQHYHQGYY DIP protein Q00839 original sequence version: 122 5D4EC4188436831F hnrpl_human Heterogeneous nuclear ribonucleoprotein L HNRPL HNRNPL P/OKcl.14 protein protein human Homo sapiens Human MSRRLLPRAEKRRRRLEQRQQPDEQRRRSGAMVKMAAAGGGGGGGRYYGGGSEGGRAPKRLKTDNAGDQHGGGGGGGGGAGAAGGGGGGENYDDPHKTPASPVVHIRGLIDGVVEADLVEALQEFGPISYVVVMPKKRQALVEFEDVLGACNAVNYAADNQIYIAGHPAFVNYSTSQKISRPGDSDDSRSVNSVLLFTILNPIYSITTDVLYTICNPCGPVQRIVIFRKNGVQAMVEFDSVQSAQRAKASLNGADIYSGCCTLKIEYAKPTRLNVFKNDQDTWDYTNPNLSGQGDPGSNPNKRQRQPPLLGDHPAEYGGPHGGYHSHYHDEGYGPPPPHYEGRRMGPPVGGHRRGPSRYGPQYGHPPPPPPPPEYGPHADSPVLMVYGLDQSKMNCDRVFNVFCLYGNVEKVKFMKSKPGAAMVEMADGYAVDRAITHLNNNFMFGQKLNVCVSKQPAIMPGQSYGLEDGSCSYKDFSESRNNRFSTPEQAAKNRIQHPSNVLHFFNAPLEVTEENFFEICDELGVKRPSSVKVFSGKSERSSSGLLEWESKSDALETLGFLNHYQMKNPNGPYPYTLKLCFSTAQHAS DIP protein P14866 original sequence version: 145 31EEB51AF1C65F83 zn827_human Zinc finger protein 827 ZNF827 protein protein human Homo sapiens Human MPRRKQEQPKRLPSHVSRQEEAEGELSEGEHWYGNSSETPSEASYGEVQENYKLSLEDRIQEQSTSPDTSLGSTTPSSHTLELVALDSEVLRDSLQCQDHLSPGVSSLCDDDPGSNKPLSSNLRRLLEAGSLKLDAAATANGRVESPVNVGSNLSFSPPSHHAQQLSVLARKLAEKQEQNDQYTPSNRFIWNQGKWLPNSTTTCSLSPDSAILKLKAAANAVLQDKSLTRTEETMRFESFSSPFSSQSASSTLAALSKKVSERSLTPGQEHPPPASSFLSLASMTSSAALLKEVAARAAGSLLAEKSSLLPEDPLPPPPSEKKPEKVTPPPPPPPPPPPPPPPQSLELLLLPVPKGRVSKPSNSASEEESGKPFQCPICGLVIKRKSYWKRHMVIHTGLKSHQCPLCPFRCARKDNLKSHMKVHQHQDRGETFQCQLCPFTSSRHFSLKLHMRCHQHFLRTEAKVKEEIPDPDVKGSPHLSDSACLGQQREGGGTELVGTMMTSNTPERTSQGGAGVSPLLVKEEPKEDNGLPTSFTLNAADRPANHTKLKDPSEYVANSASALFSQDISVKMASDFLMKLSAANQKEPMNLNFKVKEEPKEGESLSTTLPRSSYVFSPESEVSAPGVSEDALKPQEGKGSVLRRDVSVKAASELLMKLSAESYKETQMVKIKEEPMEVDIQDSHVSISPSRNVGYSTLIGREKTEPLQKMPEGRVPPERNLFSQDISVKMASELLFQLSEKVSKEHNHTKENTIRTTTSPFFSEDTFRQSPFTSNSKELLPSDSVLHGRISAPETEKIVLEAGNGLPSWKFNDQLFPCDVCGKVFGRQQTLSRHLSLHTEERKYKCHLCPYAAKCRANLNQHLTVHSVKLVSTDTEDIVSAVTSEGSDGKKHPYYYSCHVCGFETELNVQFVSHMSLHVDKEQWMFSICCTACDFVTMEEAEIKTHIGTKHTGEDRKTPSESNSPSSSSLSALSDSANSKDDSDGSQKNKGGNNLLVISVMPGSQPSLNSEEKPEKGFECVFCNFVCKTKNMFERHLQIHLITRMFECDVCHKFMKTPEQLLEHKKCHTVPTGGLNSGQW DIP protein Q17R98 original sequence version: 79 BA32DF19FE700EFA hnrnpk-hnrnpu-1 3 6 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association fig. 5d To assess interactions between hnRNP K, -L and -U, and their potential dependence on circZNF827, we performed co-immunoprecipitation experiments using FLAG-tagged hnRNP K, -L, and -U, and subsequently probed for their interaction with endogenous proteins (Figure 5D) in HEK293 Flp-in cells either overexpressing circZNF827 or not. hnRNP K co-immunoprecipitates both hnRNP U and hnRNP L (long isoform), but these interactions remain unaffected by increased expression of circZNF827 (Figure 5D). hnrnpk-10 3 6 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association fig. 5d To assess interactions between hnRNP K, -L and -U, and their potential dependence on circZNF827, we performed co-immunoprecipitation experiments using FLAG-tagged hnRNP K, -L, and -U, and subsequently probed for their interaction with endogenous proteins (Figure 5D) in HEK293 Flp-in cells either overexpressing circZNF827 or not. hnRNP K co-immunoprecipitates both hnRNP U and hnRNP L (long isoform), but these interactions remain unaffected by increased expression of circZNF827 (Figure 5D). hnrnpk-hnrnpl-1 3 7 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 4 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association fig.5e In accordance with these findings, immunoprecipitation of endogenous hnRNP K and -L proteins in HEK293 Flp-in cells, confirmed that a hnRNP L/hnRNP K complex can be detected although only a small fraction of the total hnRNP K/- L populations co-immunoprecipitate the other (Figure 5E, left). This complex was not affected by overexpression of circZNF827 (Figure 5E, right). hnrnpk-11 3 7 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level 4 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association fig.5e In accordance with these findings, immunoprecipitation of endogenous hnRNP K and -L proteins in HEK293 Flp-in cells, confirmed that a hnRNP L/hnRNP K complex can be detected although only a small fraction of the total hnRNP K/- L populations co-immunoprecipitate the other (Figure 5E, left). This complex was not affected by overexpression of circZNF827 (Figure 5E, right). hnrnpk-hnrnpl-2 2 7 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell 4 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization fig. 5g hnrnpk-hnrnpu-2 3 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 4 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed association association fig. 5d To assess interactions between hnRNP K, -L and -U, and their potential dependence on circZNF827, we performed co-immunoprecipitation experiments using FLAG-tagged hnRNP K, -L, and -U, and subsequently probed for their interaction with endogenous proteins (Figure 5D) in HEK293 Flp-in cells either overexpressing circZNF827 or not. hnRNP K co-immunoprecipitates both hnRNP U and hnRNP L (long isoform), but these interactions remain unaffected by increased expression of circZNF827 (Figure 5D). hnrnpk-12 1 7 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level 4 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level 8 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level association association fig. 6e To address whether ZNF827 and hnRNP K interact, we next performed ZNF827 immunoprecipitation and found it to strongly associate with hnRNP K and to a lesser extent with hnRNP L in nucleoplasmic extracts (Figure 6E, left). Upon sonication of the remainder from the Triton X-100 extracted cleared lysates (chromatin enriched), we observed an even stronger association of hnRNP K with ZNF827, further suggesting that the complex is chromatin bound (Figure 6E, right). znf827-hnrnpk-1 1 7 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level 4 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level 8 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level association association fig. 6e To address whether ZNF827 and hnRNP K interact, we next performed ZNF827 immunoprecipitation and found it to strongly associate with hnRNP K and to a lesser extent with hnRNP L in nucleoplasmic extracts (Figure 6E, left). Upon sonication of the remainder from the Triton X-100 extracted cleared lysates (chromatin enriched), we observed an even stronger association of hnRNP K with ZNF827, further suggesting that the complex is chromatin bound (Figure 6E, right).