UniProt UniProt http://www.uniprot.org larue-2012-1 Interaction of the HIV-1 intasome with transportin 3 protein (TNPO3 or TRN-SR2). in vitro In vitro pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Larue R., Gupta K., Wuensch C., Shkriabai N., Kessl J.J., Danhart E., Feng L., Taltynov O., Christ F., Van Duyne G.D., Debyser Z., Foster M.P., Kvaratskhelia M. Virus - Publications including interactions involving viral proteins HIV-1 proteins shave been mapped to the UniProt HIV-1 reference strain. Exp involving recombinant CypA not included as origin of CypA cannot be established from the publication. FLAG-tagged HIV-1 IN according ref 25 (pubmed 22437836) kvaratshkelia.1@osu.edu J. Biol. Chem. (0021-9258) 2012 imex curation Only protein-protein interactions larue-2012-8 Interaction of the HIV-1 intasome with transportin 3 protein (TNPO3 or TRN-SR2). in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip weight by comassie molecular weight estimation by coomasie staining weight by comassie Larue R., Gupta K., Wuensch C., Shkriabai N., Kessl J.J., Danhart E., Feng L., Taltynov O., Christ F., Van Duyne G.D., Debyser Z., Foster M.P., Kvaratskhelia M. Virus - Publications including interactions involving viral proteins In exp involving LEDGF/p75 FLAG-IN was preincubated with increasing concentrations of LEDF/p75 and then TPO3 was added. HIV-1 proteins shave been mapped to the UniProt HIV-1 reference strain. Exp involving recombinant CypA not included as origin of CypA cannot be established from the publication. FLAG-tagged HIV-1 IN according ref 25 (pubmed 22437836) kvaratshkelia.1@osu.edu J. Biol. Chem. (0021-9258) 2012 imex curation Only protein-protein interactions larue-2012-2 Interaction of the HIV-1 intasome with transportin 3 protein (TNPO3 or TRN-SR2). in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Larue R., Gupta K., Wuensch C., Shkriabai N., Kessl J.J., Danhart E., Feng L., Taltynov O., Christ F., Van Duyne G.D., Debyser Z., Foster M.P., Kvaratskhelia M. Virus - Publications including interactions involving viral proteins Stable synaptic complexes (SSC; a tetramer of IN associated with a pair of viral DNA ends) have been mixed with purified TNPO3 and passed through a size exclusion spin column; eluate western-blotted. HIV-1 proteins shave been mapped to the UniProt HIV-1 reference strain. Exp involving recombinant CypA not included as origin of CypA cannot be established from the publication. FLAG-tagged HIV-1 IN according ref 25 (pubmed 22437836) kvaratshkelia.1@osu.edu J. Biol. Chem. (0021-9258) 2012 imex curation Only protein-protein interactions larue-2012-3 Interaction of the HIV-1 intasome with transportin 3 protein (TNPO3 or TRN-SR2). in vitro In vitro pull down pull down fingerprinting peptide massfingerprinting fingerprinting Larue R., Gupta K., Wuensch C., Shkriabai N., Kessl J.J., Danhart E., Feng L., Taltynov O., Christ F., Van Duyne G.D., Debyser Z., Foster M.P., Kvaratskhelia M. Virus - Publications including interactions involving viral proteins IN CTD was incubated with or without TNPO3 and treated with NHS-biotin. To identify IN residues essential for high affinity binding with TNPO3, free IN and the IN–TNPO3 complex are subjected to treatment by small covalent modifiers. Surface residues in free IN and the complex are susceptible to modification, while the interacting amino acids in the complex are shielded from modification. HIV-1 proteins shave been mapped to the UniProt HIV-1 reference strain. Exp involving recombinant CypA not included as origin of CypA cannot be established from the publication. FLAG-tagged HIV-1 IN according ref 25 (pubmed 22437836) kvaratshkelia.1@osu.edu J. Biol. Chem. (0021-9258) 2012 imex curation Only protein-protein interactions larue-2012-4 Interaction of the HIV-1 intasome with transportin 3 protein (TNPO3 or TRN-SR2). in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Larue R., Gupta K., Wuensch C., Shkriabai N., Kessl J.J., Danhart E., Feng L., Taltynov O., Christ F., Van Duyne G.D., Debyser Z., Foster M.P., Kvaratskhelia M. Virus - Publications including interactions involving viral proteins Stable synaptic complexes (SSC; a tetramer of IN associated with a pair of viral DNA ends) have been mixed with purified TNPO3 and passed through a size exclusion spin column; eluate western-blotted. HIV-1 proteins shave been mapped to the UniProt HIV-1 reference strain. Exp involving recombinant CypA not included as origin of CypA cannot be established from the publication. FLAG-tagged HIV-1 IN according ref 25 (pubmed 22437836) kvaratshkelia.1@osu.edu J. Biol. Chem. (0021-9258) 2012 imex curation Only protein-protein interactions larue-2012-5 Interaction of the HIV-1 intasome with transportin 3 protein (TNPO3 or TRN-SR2). in vitro In vitro saxs x ray scattering saxs predetermined predetermined participant predetermined Larue R., Gupta K., Wuensch C., Shkriabai N., Kessl J.J., Danhart E., Feng L., Taltynov O., Christ F., Van Duyne G.D., Debyser Z., Foster M.P., Kvaratskhelia M. Virus - Publications including interactions involving viral proteins HIV-1 proteins shave been mapped to the UniProt HIV-1 reference strain. Exp involving recombinant CypA not included as origin of CypA cannot be established from the publication. FLAG-tagged HIV-1 IN according ref 25 (pubmed 22437836) kvaratshkelia.1@osu.edu J. Biol. Chem. (0021-9258) 2012 imex curation Only protein-protein interactions larue-2012-6 Interaction of the HIV-1 intasome with transportin 3 protein (TNPO3 or TRN-SR2). in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Larue R., Gupta K., Wuensch C., Shkriabai N., Kessl J.J., Danhart E., Feng L., Taltynov O., Christ F., Van Duyne G.D., Debyser Z., Foster M.P., Kvaratskhelia M. Virus - Publications including interactions involving viral proteins HIV-1 proteins shave been mapped to the UniProt HIV-1 reference strain. Exp involving recombinant CypA not included as origin of CypA cannot be established from the publication. FLAG-tagged HIV-1 IN according ref 25 (pubmed 22437836) kvaratshkelia.1@osu.edu J. Biol. Chem. (0021-9258) 2012 imex curation Only protein-protein interactions larue-2012-7 Interaction of the HIV-1 intasome with transportin 3 protein (TNPO3 or TRN-SR2). in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Larue R., Gupta K., Wuensch C., Shkriabai N., Kessl J.J., Danhart E., Feng L., Taltynov O., Christ F., Van Duyne G.D., Debyser Z., Foster M.P., Kvaratskhelia M. Virus - Publications including interactions involving viral proteins Multiangle Light Scattering for detection. HIV-1 proteins shave been mapped to the UniProt HIV-1 reference strain. Exp involving recombinant CypA not included as origin of CypA cannot be established from the publication. FLAG-tagged HIV-1 IN according ref 25 (pubmed 22437836) kvaratshkelia.1@osu.edu J. Biol. Chem. (0021-9258) 2012 imex curation Only protein-protein interactions p04585-pro_0000042447 Integrase gag-pol Pr160Gag-Pol protein protein hv1h2 Human immunodeficiency virus type 1 group M subtype B (isolate HXB2) (HIV-1) HIV-1 FLDGIDKAQDEHEKYHSNWRAMASDFNLPPVVAKEIVASCDKCQLKGEAMHGQVDCSPGIWQLDCTHLEGKVILVAVHVASGYIEAEVIPAETGQETAYFLLKLAGRWPVKTIHTDNGSNFTGATVRAACWWAGIKQEFGIPYNPQSQGVVESMNKELKKIIGQVRDQAEHLKTAVQMAVFIHNFKRKGGIGGYSAGERIVDIIATDIQTKELQKQITKIQNFRVYYRDSRNPLWKGPAKLLWKGEGAVVIQDNSDIKVVPRRKAKIIRDYGKQMAGDDCVASRQDED 1148 1435 tnpo3_human Transportin-3 TNPO3 IPO12 Transportin-SR Importin-12 protein protein human Homo sapiens Human MEGAKPTLQLVYQAVQALYHDPDPSGKERASFWLGELQRSVHAWEISDQLLQIRQDVESCYFAAQTMKMKIQTSFYELPTDSHASLRDSLLTHIQNLKDLSPVIVTQLALAIADLALQMPSWKGCVQTLVEKYSNDVTSLPFLLEILTVLPEEVHSRSLRIGANRRTEIIEDLAFYSSTVVSLLMTCVEKAGTDEKMLMKVFRCLGSWFNLGVLDSNFMANNKLLALLFEVLQQDKTSSNLHEAASDCVCSALYAIENVETNLPLAMQLFQGVLTLETAYHMAVAREDLDKVLNYCRIFTELCETFLEKIVCTPGQGLGDLRTLELLLICAGHPQYEVVEISFNFWYRLGEHLYKTNDEVIHGIFKAYIQRLLHALARHCQLEPDHEGVPEETDDFGEFRMRVSDLVKDLIFLIGSMECFAQLYSTLKEGNPPWEVTEAVLFIMAAIAKSVDPENNPTLVEVLEGVVRLPETVHTAVRYTSIELVGEMSEVVDRNPQFLDPVLGYLMKGLCEKPLASAAAKAIHNICSVCRDHMAQHFNGLLEIARSLDSFLLSPEAAVGLLKGTALVLARLPLDKITECLSELCSVQVMALKKLLSQEPSNGISSDPTVFLDRLAVIFRHTNPIVENGQTHPCQKVIQEIWPVLSETLNKHRADNRIVERCCRCLRFAVRCVGKGSAALLQPLVTQMVNVYHVHQHSCFLYLGSILVDEYGMEEGCRQGLLDMLQALCIPTFQLLEQQNGLQNHPDTVDDLFRLATRFIQRSPVTLLRSQVVIPILQWAIASTTLDHRDANCSVMRFLRDLIHTGVANDHEEDFELRKELIGQVMNQLGQQLVSQLLHTCCFCLPPYTLPDVAEVLWEIMQVDRPTFCRWLENSLKGLPKETTVGAVTVTHKQLTDFHKQVTSAEECKQVCWALRDFTRLFR o75475-1 PC4 and SFRS1-interacting protein p75 PSIP1 PSIP1 DFS70 LEDGF PSIP2 CLL-associated antigen KW-7 Dense fine speckles 70 kDa protein Lens epithelium-derived growth factor Transcriptional coactivator p75/p52 protein protein human Homo sapiens Human MTRDFKPGDLIFAKMKGYPHWPARVDEVPDGAVKPPTNKLPIFFFGTHETAFLGPKDIFPYSENKEKYGKPNKRKGFNEGLWEIDNNPKVKFSSQQAATKQSNASSDVEVEEKETSVSKEDTDHEEKASNEDVTKAVDITTPKAARRGRKRKAEKQVETEEAGVVTTATASVNLKVSPKRGRPAATEVKIPKPRGRPKMVKQPCPSESDIITEEDKSKKKGQEEKQPKKQPKKDEEGQKEEDKPRKEPDKKEGKKEVESKRKNLAKTGVTSTSDSEEEGDDQEGEKKRKGGRNFQTAHRRNMLKGQHEKEAADRKRKQEEQMETEQQNKDEGKKPEVKKVEKKRETSMDSRLQRIHAEIKNSLKIDNLDVNRCIEALDELASLQVTMQQAQKHTEMITTLKKIRRFKVSQVIMEKSTMLYNKFKNMFLVGEGDSVITQVLNKSLAEQRQHEEANKTKDQGKKGPNKKLEKEQTGSKTLNGGSDAQDGNQPQHNGESNEDSKDNHEASTKKKPSSEERETEISLKDSTLDN Less active than isoform 2 as transcriptional coactivator, but more abundant in cells. tnpo3-gag_pol 1778381 n/a 1778389 unspecified role unspecified role prey prey n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P04585-PRO_0000042447:p.Lys240Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P04585-PRO_0000042447:p.[Arg262Glu;Arg263Glu] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P04585-PRO_0000042447:p.Lys264Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false P04585-PRO_0000042447:p.[Arg262Glu;Arg263Glu;Lys264Glu] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength 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 n/a 1778390 unspecified role unspecified role bait bait n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 3 and 5A Binding with IN C-terminal region is much stronger than with N-terminal region. psip1-gag_pol 1778381 n/a 1778391 unspecified role unspecified role bait bait n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1778389 unspecified role unspecified role prey prey An IN construct arg262glu_arg263glu_lys264glu also did interact direct interaction direct interaction false false false 5F gag_pol-tnpo3-1 1778382 n/a 1778390 unspecified role unspecified role prey prey n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q9Y5L0:p.[Arg400Glu;Arg402Glu] 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 Q9Y5L0:p.Gln761_Arg762delinsGluGlu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1778389 unspecified role unspecified role bait bait n-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false direct interaction direct interaction false false false 1D and 8A and C gag_pol-psip1-22 1778382 n/a 1778391 unspecified role unspecified role prey prey n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1778389 unspecified role unspecified role bait bait n-terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1778390 unspecified role unspecified role prey prey n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) association association false false false 2C gag_pol-tnpo3-2 1778383 n/a 1778389 unspecified role unspecified role neutral component neutral component n/a 1778390 unspecified role unspecified role neutral component neutral component n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) physical association physical association false false false 2B gag_pol-tnpo3-3 1778384 n/a 1778389 unspecified role unspecified role unspecified role unspecified role n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag certain certain sequence position certain certain certain sequence position certain false n/a 1778390 unspecified role unspecified role unspecified role unspecified role ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 4 and Table S2 Tryptic peptide 264-269 with K-264 and K-266 biotinylated has a diminished peak upon IN CTD binding to TPNO3. tnpo3-gag_pol-1 1778385 n/a 1778390 unspecified role unspecified role bait bait n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1778389 unspecified role unspecified role prey prey P04585-PRO_0000042447:p.Arg262_Lys264delinsGluGluGlu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false direct interaction direct interaction false false false 5D tnpo3-2 1778386 n/a 1778390 unspecified role unspecified role neutral component neutral component Stoichiometry: 2.0 direct interaction direct interaction false false false 7 tnpo3-2 1778387 n/a 1778390 unspecified role unspecified role neutral component neutral component n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false 1778387 6E and 6D tnpo3-2 1778388 n/a 1778390 unspecified role unspecified role neutral component neutral component n-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false 6C