MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>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 http://mint.bio.uniroma2.it/mint chang-2013b-4 Chtop is a component of the dynamic TREX mRNA export complex. in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Chang C.T., Hautbergue G.M., Walsh M.J., Viphakone N., van Dijk T.B., Philipsen S., Wilson S.A. stuart.wilson@sheffield.ac.uk EMBO J. (0261-4189) 2013 Only protein-protein interactions imex curation chang-2013b-5 Chtop is a component of the dynamic TREX mRNA export complex. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Chang C.T., Hautbergue G.M., Walsh M.J., Viphakone N., van Dijk T.B., Philipsen S., Wilson S.A. stuart.wilson@sheffield.ac.uk EMBO J. (0261-4189) 2013 Only protein-protein interactions imex curation chang-2013b-1 Chtop is a component of the dynamic TREX mRNA export complex. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Chang C.T., Hautbergue G.M., Walsh M.J., Viphakone N., van Dijk T.B., Philipsen S., Wilson S.A. stuart.wilson@sheffield.ac.uk EMBO J. (0261-4189) 2013 Only protein-protein interactions imex curation chang-2013b-3 Chtop is a component of the dynamic TREX mRNA export complex. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Chang C.T., Hautbergue G.M., Walsh M.J., Viphakone N., van Dijk T.B., Philipsen S., Wilson S.A. stuart.wilson@sheffield.ac.uk EMBO J. (0261-4189) 2013 Only protein-protein interactions imex curation chang-2013b-2 Chtop is a component of the dynamic TREX mRNA export complex. in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Chang C.T., Hautbergue G.M., Walsh M.J., Viphakone N., van Dijk T.B., Philipsen S., Wilson S.A. stuart.wilson@sheffield.ac.uk EMBO J. (0261-4189) 2013 Only protein-protein interactions imex curation chang-2013b-6 Chtop is a component of the dynamic TREX mRNA export complex. human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Chang C.T., Hautbergue G.M., Walsh M.J., Viphakone N., van Dijk T.B., Philipsen S., Wilson S.A. stuart.wilson@sheffield.ac.uk EMBO J. (0261-4189) 2013 Only protein-protein interactions imex curation dx39b_human Spliceosome RNA helicase DDX39B UAP56 DDX39B DEAD box protein UAP56 56 kDa U2AF65-associated protein ATP-dependent RNA helicase p47 HLA-B-associated transcript 1 protein BAT1 protein protein human Homo sapiens Human MAENDVDNELLDYEDDEVETAAGGDGAEAPAKKDVKGSYVSIHSSGFRDFLLKPELLRAIVDCGFEHPSEVQHECIPQAILGMDVLCQAKSGMGKTAVFVLATLQQLEPVTGQVSVLVMCHTRELAFQISKEYERFSKYMPNVKVAVFFGGLSIKKDEEVLKKNCPHIVVGTPGRILALARNKSLNLKHIKHFILDECDKMLEQLDMRRDVQEIFRMTPHEKQVMMFSATLSKEIRPVCRKFMQDPMEIFVDDETKLTLHGLQQYYVKLKDNEKNRKLFDLLDVLEFNQVVIFVKSVQRCIALAQLLVEQNFPAIAIHRGMPQEERLSRYQQFKDFQRRILVATNLFGRGMDIERVNIAFNYDMPEDSDTYLHRVARAGRFGTKGLAITFVSDENDAKILNDVQDRFEVNISELPDEIDISSYIEQTR chtop_human Chromatin target of PRMT1 protein HT031 PP7704 FOP CHTOP C1orf77 Friend of PRMT1 protein Small arginine- and glycine-rich protein protein protein human Homo sapiens Human MAAQSAPKVVLKSTTKMSLNERFTNMLKNKQPTPVNIRASMQQQQQLASARNRRLAQQMENRPSVQAALKLKQSLKQRLGKSNIQARLGRPIGALARGAIGGRGLPIIQRGLPRGGLRGGRATRTLLRGGMSLRGQNLLRGGRAVAPRMGLRRGGVRGRGGPGRGGLGRGAMGRGGIGGRGRGMIGRGRGGFGGRGRGRGRGRGALARPVLTKEQLDNQLDAYMSKTKGHLDAELDAYMAQTDPETND nxf1_human Nuclear RNA export factor 1 NXF1 TAP Tip-associating protein Tip-associated protein mRNA export factor TAP protein protein human Homo sapiens Human MADEGKSYSEHDDERVNFPQRKKKGRGPFRWKYGEGNRRSGRGGSGIRSSRLEEDDGDVAMSDAQDGPRVRYNPYTTRPNRRGDTWHDRDRIHVTVRRDRAPPERGGAGTSQDGTSKNWFKITIPYGRKYDKAWLLSMIQSKCSVPFTPIEFHYENTRAQFFVEDASTASALKAVNYKILDRENRRISIIINSSAPPHTILNELKPEQVEQLKLIMSKRYDGSQQALDLKGLRSDPDLVAQNIDVVLNRRSCMAATLRIIEENIPELLSLNLSNNRLYRLDDMSSIVQKAPNLKILNLSGNELKSERELDKIKGLKLEELWLDGNSLCDTFRDQSTYISAIRERFPKLLRLDGHELPPPIAFDVEAPTTLPPCKGSYFGTENLKSLVLHFLQQYYAIYDSGDRQGLLDAYHDGACCSLSIPFIPQNPARSSLAEYFKDSRNVKKLKDPTLRFRLLKHTRLNVVAFLNELPKTQHDVNSFVVDISAQTSTLLCFSVNGVFKEVDGKSRDSLRAFTRTFIAVPASNSGLCIVNDELFVRNASSEEIQRAFAMPAPTPSSSPVPTLSPEQQEMLQAFSTQSGMNLEWSQKCLQDNNWDYTRSAQAFTHLKAKGEIPEVAFMK thoc4_human THO complex subunit 4 ALYREF ALY Ally of AML-1 and LEF-1 Aly/REF export factor BEF THOC4 Transcriptional coactivator Aly/REF bZIP-enhancing factor BEF protein protein human Homo sapiens Human MADKMDMSLDDIIKLNRSQRGGRGGGRGRGRAGSQGGRGGGAQAAARVNRGGGPIRNRPAIARGAAGGGGRNRPAPYSRPKQLPDKWQHDLFDSGFGGGAGVETGGKLLVSNLDFGVSDADIQELFAEFGTLKKAAVHYDRSGRSLGTADVHFERKADALKAMKQYNGVPLDGRPMNIQLVTSQIDAQRRPAQSVNRGGMTRNRGAGGFGGGGGTRRGTRGGARGRGRGAGRNSKQQLSAEELDAQLDAYNARMDTS nxt1_human NTF2-related export protein 1 NXT1 Protein p15 protein protein human Homo sapiens Human MASVDFKTYVDQACRAAEEFVNVYYTTMDKRRRLLSRLYMGTATLVWNGNAVSGQESLSEFFEMLPSSEFQISVVDCQPVHDEATPSQTTVLVVICGSVKFEGNKQRDFNQNFILTAQASPSNTVWKIASDCFRFQDWAS uif_human UAP56-interacting factor UIF FYTTD1 Forty-two-three domain-containing protein 1 protein protein human Homo sapiens Human MNRFGTRLVGATATSSPPPKARSNENLDKIDMSLDDIIKLNRKEGKKQNFPRLNRRLLQQSGAQQFRMRVRWGIQQNSGFGKTSLNRRGRVMPGKRRPNGVITGLAARKTTGIRKGISPMNRPPLSDKNIEQYFPVLKRKANLLRQNEGQRKPVAVLKRPSQLSRKNNIPANFTRSGNKLNHQKDTRQATFLFRRGLKVQAQLNTEQLLDDVVAKRTRQWRTSTTNGGILTVSIDNPGAVQCPVTQKPRLTRTAVPSFLTKREQSDVKKVPKGVPLQFDINSVGKQTGMTLNERFGILKEQRATLTYNKGGSRFVTVG thoc2_human THO complex subunit 2 hTREX120 THOC2 CXorf3 protein protein human Homo sapiens Human MAAAAVVVPAEWIKNWEKSGRGEFLHLCRILSENKSHDSSTYRDFQQALYELSYHVIKGNLKHEQASNVLSDISEFREDMPSILADVFCILDIETNCLEEKSKRDYFTQLVLACLYLVSDTVLKERLDPETLESLGLIKQSQQFNQKSVKIKTKLFYKQQKFNLLREENEGYAKLIAELGQDLSGSITSDLILENIKSLIGCFNLDPNRVLDVILEVFECRPEHDDFFISLLESYMSMCEPQTLCHILGFKFKFYQEPNGETPSSLYRVAAVLLQFNLIDLDDLYVHLLPADNCIMDEHKREIAEAKQIVRKLTMVVLSSEKMDEREKEKEKEEEKVEKPPDNQKLGLLEALLKIGDWQHAQNIMDQMPPYYAASHKLIALAICKLIHITIEPLYRRVGVPKGAKGSPVNALQNKRAPKQAESFEDLRRDVFNMFCYLGPHLSHDPILFAKVVRIGKSFMKEFQSDGSKQEDKEKTEVILSCLLSITDQVLLPSLSLMDCNACMSEELWGMFKTFPYQHRYRLYGQWKNETYNSHPLLVKVKAQTIDRAKYIMKRLTKENVKPSGRQIGKLSHSNPTILFDYILSQIQKYDNLITPVVDSLKYLTSLNYDVLAYCIIEALANPEKERMKHDDTTISSWLQSLASFCGAVFRKYPIDLAGLLQYVANQLKAGKSFDLLILKEVVQKMAGIEITEEMTMEQLEAMTGGEQLKAEGGYFGQIRNTKKSSQRLKDALLDHDLALPLCLLMAQQRNGVIFQEGGEKHLKLVGKLYDQCHDTLVQFGGFLASNLSTEDYIKRVPSIDVLCNEFHTPHDAAFFLSRPMYAHHISSKYDELKKSEKGSKQQHKVHKYITSCEMVMAPVHEAVVSLHVSKVWDDISPQFYATFWSLTMYDLAVPHTSYEREVNKLKVQMKAIDDNQEMPPNKKKKEKERCTALQDKLLEEEKKQMEHVQRVLQRLKLEKDNWLLAKSTKNETITKFLQLCIFPRCIFSAIDAVYCARFVELVHQQKTPNFSTLLCYDRVFSDIIYTVASCTENEASRYGRFLCCMLETVTRWHSDRATYEKECGNYPGFLTILRATGFDGGNKADQLDYENFRHVVHKWHYKLTKASVHCLETGEYTHIRNILIVLTKILPWYPKVLNLGQALERRVHKICQEEKEKRPDLYALAMGYSGQLKSRKSYMIPENEFHHKDPPPRNAVASVQNGPGGGPSSSSIGSASKSDESSTEETDKSRERSQCGVKAVNKASSTTPKGNSSNGNSGSNSNKAVKENDKEKGKEKEKEKKEKTPATTPEARVLGKDGKEKPKEERPNKDEKARETKERTPKSDKEKEKFKKEEKAKDEKFKTTVPNAESKSTQEREREKEPSRERDIAKEMKSKENVKGGEKTPVSGSLKSPVPRSDIPEPEREQKRRKIDTHPSPSHSSTVKDSLIELKESSAKLYINHTPPPLSKSKEREMDKKDLDKSRERSREREKKDEKDRKERKRDHSNNDREVPPDLTKRRKEENGTMGVSKHKSESPCESPYPNEKDKEKNKSKSSGKEKGSDSFKSEKMDKISSGGKKESRHDKEKIEKKEKRDSSGGKEEKKHHKSSDKHR thoc5_human THO complex subunit 5 homolog THOC5 C22orf19 KIAA0983 Functional spliceosome-associated protein 79 NF2/meningioma region protein pK1.3 Placental protein 39.2 hTREX90 protein protein human Homo sapiens Human MSSESSKKRKPKVIRSDGAPAEGKRNRSDTEQEGKYYSEEAEVDLRDPGRDYELYKYTCQELQRLMAEIQDLKSRGGKDVAIEIEERRIQSCVHFMTLKKLNRLAHIRLKKGRDQTHEAKQKVDAYHLQLQNLLYEVMHLQKEITKCLEFKSKHEEIDLVSLEEFYKEAPPDISKAEVTMGDPHQQTLARLDWELEQRKRLAEKYRECLSNKEKILKEIEVKKEYLSSLQPRLNSIMQASLPVQEYLFMPFDQAHKQYETARHLPPPLYVLFVQATAYGQACDKTLSVAIEGSVDEAKALFKPPEDSQDDESDSDAEEEQTTKRRRPTLGVQLDDKRKEMLKRHPLSVMLDLKCKDDSVLHLTFYYLMNLNIMTVKAKVTTAMELITPISAGDLLSPDSVLSCLYPGDHGKKTPNPANQYQFDKVGILTLSDYVLELGHPYLWVQKLGGLHFPKEQPQQTVIADHSLSASHMETTMKLLKTRVQSRLALHKQFASLEHGIVPVTSDCQYLFPAKVVSRLVKWVTVAHEDYMELHFTKDIVDAGLAGDTNLYYMALIERGTAKLQAAVVLNPGYSSIPPVFQLCLNWKGEKTNSNDDNIRAMEGEVNVCYKELCGPWPSHQLLTNQLQRLCVLLDVYLETESHDDSVEGPKEFPQEKMCLRLFRGPSRMKPFKYNHPQGFFSHR thoc1_human THO complex subunit 1 hTREX84 THOC1 HPR1 Nuclear matrix protein p84 protein protein human Homo sapiens Human MSPTPPLFSLPEARTRFTKSTREALNNKNIKPLLSTFSQVPGSENEKKCTLDQAFRGILEEEIINHSSCENVLAIISLAIGGVTEGICTASTPFVLLGDVLDCLPLDQCDTIFTFVEKNVATWKSNTFYSAGKNYLLRMCNDLLRRLSKSQNTVFCGRIQLFLARLFPLSEKSGLNLQSQFNLENVTVFNTNEQESTLGQKHTEDREEGMDVEEGEMGDEEAPTTCSIPIDYNLYRKFWSLQDYFRNPVQCYEKISWKTFLKYSEEVLAVFKSYKLDDTQASRKKMEELKTGGEHVYFAKFLTSEKLMDLQLSDSNFRRHILLQYLILFQYLKGQVKFKSSNYVLTDEQSLWIEDTTKSVYQLLSENPPDGERFSKMVEHILNTEENWNSWKNEGCPSFVKERTSDTKPTRIIRKRTAPEDFLGKGPTKKILMGNEELTRLWNLCPDNMEACKSETREHMPTLEEFFEEAIEQADPENMVENEYKAVNNSNYGWRALRLLARRSPHFFQPTNQQFKSLPEYLENMVIKLAKELPPPSEEIKTGEDEDEEDNDALLKENESPDVRRDKPVTGEQIEVFANKLGEQWKILAPYLEMKDSEIRQIECDSEDMKMRAKQLLVAWQDQEGVHATPENLINALNKSGLSDLAESLTNDNETNS srsf2_human Serine/arginine-rich splicing factor 2 Splicing factor SC35 Splicing component, 35 kDa Protein PR264 SRSF2 SFRS2 Splicing factor, arginine/serine-rich 2 protein protein human Homo sapiens Human MSYGRPPPDVEGMTSLKVDNLTYRTSPDTLRRVFEKYGRVGDVYIPRDRYTKESRGFAFVRFHDKRDAEDAMDAMDGAVLDGRELRVQMARYGRPPDSHHSRRGPPPRRYGGGGYGRRSRSPRRRRRSRSRSRSRSRSRSRSRYSRSKSRSRTRSRSRSTSKSRSARRSKSKSSSVSRSRSRSRSRSRSRSPPPVSKRESKSRSRSKSPPKSPEEEGAVSS ddx39b-chtop-2 1476423 n/a 1476429 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1476430 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false f3e nxf1-chtop-2 1476423 n/a 1476430 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1476431 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false f3e alyref-chtop-1 1476423 n/a 1476430 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1476432 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false f3e chtop-alyref-1 1476423 n/a 1476433 predetermined predetermined participant predetermined unspecified role unspecified role unspecified role unspecified role purified purified n/a 1476431 weight by comassie molecular weight estimation by coomasie staining weight by comassie unspecified role unspecified role unspecified role unspecified role purified purified tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1476430 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate 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 false association association false false false f4c f4e We investigated what impact Alyref had on the interaction of Chtop with Nxf1 (Figure 4C) and found that increasing amounts of GB1-Alyref bound to Nxf1, stimulated the interaction of FLAG-Chtop with Nxf1. In contrast, FLAGUif, which binds aa 1–198 of Nxf1, was efficiently displaced by Gb1-Alyref in this assay. Together, these data suggest that Alyref stimulates Chtop binding to Nxf1, though we have not observed a reciprocal stimulation of Alyref binding to Nxf1 by Chtop. To confirm that Chtop and Thoc5 utilise a common binding site on Nxf1, we examined the effects of increasing amounts of Thoc5 on the Nxf1:Chtop interaction and found that Thoc5 efficiently displaces Chtop from Nxf1 (Figure 4E). chtop-nxf1 1476423 n/a 1476433 predetermined predetermined participant predetermined unspecified role unspecified role unspecified role unspecified role purified purified n/a 1476434 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate 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 false n/a 1476431 weight by comassie molecular weight estimation by coomasie staining weight by comassie unspecified role unspecified role unspecified role unspecified role purified purified tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false association association false false false f4c We investigated what impact Alyref had on the interaction of Chtop with Nxf1 (Figure 4C) and found that increasing amounts of GB1-Alyref bound to Nxf1, stimulated the interaction of FLAG-Chtop with Nxf1. In contrast, FLAGUif, which binds aa 1–198 of Nxf1, was efficiently displaced by Gb1-Alyref in this assay. Together, these data suggest that Alyref stimulates Chtop binding to Nxf1, though we have not observed a reciprocal stimulation of Alyref binding to Nxf1 by Chtop. chtop-nxf1-1 1476423 n/a 1476430 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate 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 false n/a 1476431 weight by comassie molecular weight estimation by coomasie staining weight by comassie unspecified role unspecified role unspecified role unspecified role purified purified Q9UBU9:p.[Leu450Ala;Arg451Ala;Phe452Ala;Arg453Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false range binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false n/a 1476433 predetermined predetermined participant predetermined unspecified role unspecified role unspecified role unspecified role purified purified association association false false false f4d chtop-nxf1-2 1476423 n/a 1476433 predetermined predetermined participant predetermined unspecified role unspecified role unspecified role unspecified role purified purified n/a 1476430 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate 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 false n/a 1476431 weight by comassie molecular weight estimation by coomasie staining weight by comassie unspecified role unspecified role unspecified role unspecified role purified purified Q9UBU9:p.[Lys456Ala;His457Ala;Thr458Ala;Arg459Ala] mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false range binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false association association false false false f4d alyref-chtop-2 1476424 n/a 1476430 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476432 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell 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 false n/a 1476435 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476429 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476436 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476437 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false f8a To address this, we IPed FLAG-Nxf1 and FLAG-Alyref from cells and gently eluted the IP from beads using FLAG peptide. A proportion of the IP was subjected to western analysis for TREX components (Figure 8A). In the FLAG-Alyref IP, we detected multiple TREX components including Uap56. FLAG-Nxf1 associated with multiple TREX components except Uap56, as expected since Uap56 is displaced from Alyref by Nxf1 (Hautbergue et al, 2008). We then took the eluate from the FLAG-Nxf1 IP and subjected it to a second round of IP with control (anti-HA), Alyref, Chtop and Thoc5 antibodies and analysed these IPs for TREX components. The control antibody used in the secondary IP failed to immunoprecipitate with any TREX component. In contrast, secondary IPs with Chtop, Alyref and Thoc5 antibodies revealed interaction with multiple TREX components but no Uap56 was detected as expected. These data are consistent with Nxf1, Chtop, Thoc5 and other TREX components existing as a single complex in vivo. alyref-chtop-3 1476424 n/a 1476431 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell 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 false n/a 1476437 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476436 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476432 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476435 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476430 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false f8a To address this, we IPed FLAG-Nxf1 and FLAG-Alyref from cells and gently eluted the IP from beads using FLAG peptide. A proportion of the IP was subjected to western analysis for TREX components (Figure 8A). In the FLAG-Alyref IP, we detected multiple TREX components including Uap56. FLAG-Nxf1 associated with multiple TREX components except Uap56, as expected since Uap56 is displaced from Alyref by Nxf1 (Hautbergue et al, 2008). We then took the eluate from the FLAG-Nxf1 IP and subjected it to a second round of IP with control (anti-HA), Alyref, Chtop and Thoc5 antibodies and analysed these IPs for TREX components. The control antibody used in the secondary IP failed to immunoprecipitate with any TREX component. In contrast, secondary IPs with Chtop, Alyref and Thoc5 antibodies revealed interaction with multiple TREX components but no Uap56 was detected as expected. These data are consistent with Nxf1, Chtop, Thoc5 and other TREX components existing as a single complex in vivo. chtop-alyref-3 1476424 n/a 1476436 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476437 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476429 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476432 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476430 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell 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 false association association false false false sf1 To address this, we IPed FLAG-Nxf1 and FLAG-Alyref from cells and gently eluted the IP from beads using FLAG peptide. A proportion of the IP was subjected to western analysis for TREX components (Figure 8A). In the FLAG-Alyref IP, we detected multiple TREX components including Uap56. FLAG-Nxf1 associated with multiple TREX components except Uap56, as expected since Uap56 is displaced from Alyref by Nxf1 (Hautbergue et al, 2008). We then took the eluate from the FLAG-Nxf1 IP and subjected it to a second round of IP with control (anti-HA), Alyref, Chtop and Thoc5 antibodies and analysed these IPs for TREX components. The control antibody used in the secondary IP failed to immunoprecipitate with any TREX component. In contrast, secondary IPs with Chtop, Alyref and Thoc5 antibodies revealed interaction with multiple TREX components but no Uap56 was detected as expected. These data are consistent with Nxf1, Chtop, Thoc5 and other TREX components existing as a single complex in vivo. alyref-chtop-4 1476424 n/a 1476432 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey partially purified partially purified n/a 1476436 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey partially purified partially purified n/a 1476435 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey partially purified partially purified n/a 1476437 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey partially purified partially purified n/a 1476431 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey partially purified partially purified 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 false n/a 1476430 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait partially purified partially purified association association false false false f8a chtop-alyref-2 1476424 n/a 1476437 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey partially purified partially purified n/a 1476435 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey partially purified partially purified n/a 1476432 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait partially purified partially purified n/a 1476436 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey partially purified partially purified n/a 1476431 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey partially purified partially purified 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 false n/a 1476430 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey partially purified partially purified association association false false false f8a alyref-chtop-5 1476424 n/a 1476435 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey partially purified partially purified n/a 1476431 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey partially purified partially purified 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 false n/a 1476432 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey partially purified partially purified n/a 1476436 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait partially purified partially purified n/a 1476430 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey partially purified partially purified n/a 1476437 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey partially purified partially purified association association false false false f8a alyref-chtop 1476425 n/a 1476437 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476429 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476430 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476432 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476436 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false f1c thoc1-alyref 1476425 n/a 1476437 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476432 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476429 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476430 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476436 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false f1c chtop-alyref 1476425 n/a 1476432 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476437 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476429 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476430 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476436 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false f1c nxf1-chtop 1476425 n/a 1476430 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476431 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false f3d ddx39b-chtop 1476426 n/a 1476429 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1476430 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false f3c ddx39b-chtop-1 1476426 n/a 1476430 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476431 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false f3c nxf1-chtop-1 1476426 n/a 1476430 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1476432 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false f3c nxt1-chtop 1476426 n/a 1476433 weight by comassie molecular weight estimation by coomasie staining weight by comassie unspecified role unspecified role bait bait purified purified tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1476430 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey purified purified tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1476431 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey purified purified tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false f4f chtop-ddx39b 1476427 n/a 1476430 weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated n/a 1476429 weight by comassie molecular weight estimation by coomasie staining weight by comassie unspecified role unspecified role bait bait purified purified tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false direct interaction direct interaction false false false f1b alyref-nxf1 1476427 n/a 1476431 weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated range binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false n/a 1476432 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false direct interaction direct interaction false false false f4b alyref-nxf1-1 1476427 n/a 1476431 weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated range binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false n/a 1476430 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false direct interaction direct interaction false false false f4b chtop-srsf2 1476428 n/a 1476430 immunostaining immunostaining unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell n/a 1476438 immunostaining immunostaining unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization false false false sf2 In common with several other mRNA export factors, Chtop shows partial colocalisation with the splicing factor Srsf2 in nuclear speckles (Supplementary Figure S2).