IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ Influenza virus PB1-F2 protein induces cell death through mitochondrial ANT3 and VDAC1. Influenza virus PB1-F2 protein induces cell death through mitochondrial ANT3 and VDAC1. PLoS pathogens 2005 imex curation Zamarin D., García-Sastre A., Xiao X., Wang R., Palese P. Peter.Palese@mssm.edu 30-Aug-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Zamarin D., García-Sastre A., Xiao X., Wang R., Palese P. Peter.Palese@mssm.edu Accepted 2019-AUG-12 AT 08:42 BST AT 08:42 BST by MGT PLoS pathogens 2005 Only protein-protein interactions imex curation imex curation Influenza virus PB1-F2 protein induces cell death through mitochondrial ANT3 and VDAC1. Influenza virus PB1-F2 protein induces cell death through mitochondrial ANT3 and VDAC1. PLoS pathogens 2005 imex curation Zamarin D., García-Sastre A., Xiao X., Wang R., Palese P. Peter.Palese@mssm.edu 30-Aug-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 anti tag western anti tag western blot anti tag western Zamarin D., García-Sastre A., Xiao X., Wang R., Palese P. Peter.Palese@mssm.edu Accepted 2019-AUG-12 AT 08:42 BST AT 08:42 BST by MGT PLoS pathogens 2005 Only protein-protein interactions imex curation imex curation Influenza virus PB1-F2 protein induces cell death through mitochondrial ANT3 and VDAC1. Influenza virus PB1-F2 protein induces cell death through mitochondrial ANT3 and VDAC1. PLoS pathogens 2005 imex curation Zamarin D., García-Sastre A., Xiao X., Wang R., Palese P. Peter.Palese@mssm.edu 30-Aug-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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. pull down pull down sequence tag sequence tag identification sequence tag Zamarin D., García-Sastre A., Xiao X., Wang R., Palese P. Beta Tubulin identified by mass spectrometry (Fig.5A) could not be mapped to a unique UniprotAc and was not captured. Peter.Palese@mssm.edu Accepted 2019-AUG-12 AT 08:42 BST AT 08:42 BST by MGT PLoS pathogens 2005 Only protein-protein interactions imex curation imex curation Influenza virus PB1-F2 protein induces cell death through mitochondrial ANT3 and VDAC1. Influenza virus PB1-F2 protein induces cell death through mitochondrial ANT3 and VDAC1. PLoS pathogens 2005 imex curation Zamarin D., García-Sastre A., Xiao X., Wang R., Palese P. Peter.Palese@mssm.edu 30-Aug-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 anti tag western anti tag western blot anti tag western Zamarin D., García-Sastre A., Xiao X., Wang R., Palese P. Peter.Palese@mssm.edu Accepted 2019-AUG-12 AT 08:42 BST AT 08:42 BST by MGT PLoS pathogens 2005 Only protein-protein interactions imex curation imex curation pb1f2_i34a1 Protein PB1-F2 PB1 protein protein i34a1 Influenza A virus (strain A/Puerto Rico/8/1934 H1N1) MGQEQDTPWILSTGHISTQKRQDGQQTPKLEHRNSTRLMGHCQKTMNQVVMPKQIVYWKQWLSLRNPILVFLKTRVLKRWRLFSKHE 832A7E34E350B454 adt3_human ADP/ATP translocase 3 SLC25A6 ANT3 CDABP0051 Adenine nucleotide translocator 3 ADP,ATP carrier protein 3 Solute carrier family 25 member 6 ADP,ATP carrier protein, isoform T2 AAC3 protein protein human Homo sapiens Human MTEQAISFAKDFLAGGIAAAISKTAVAPIERVKLLLQVQHASKQIAADKQYKGIVDCIVRIPKEQGVLSFWRGNLANVIRYFPTQALNFAFKDKYKQIFLGGVDKHTQFWRYFAGNLASGGAAGATSLCFVYPLDFARTRLAADVGKSGTEREFRGLGDCLVKITKSDGIRGLYQGFSVSVQGIIIYRAAYFGVYDTAKGMLPDPKNTHIVVSWMIAQTVTAVAGVVSYPFDTVRRRMMMQSGRKGADIMYTGTVDCWRKIFRDEGGKAFFKGAWSNVLRGMGGAFVLVLYDELKKVI 18534E9F0E49672F vdac1_human Voltage-dependent anion-selective channel protein 1 VDAC1 VDAC Outer mitochondrial membrane protein porin 1 Plasmalemmal porin Porin 31HL Porin 31HM protein protein human Homo sapiens Human MAVPPTYADLGKSARDVFTKGYGFGLIKLDLKTKSENGLEFTSSGSANTETTKVTGSLETKYRWTEYGLTFTEKWNTDNTLGTEITVEDQLARGLKLTFDSSFSPNTGKKNAKIKTGYKREHINLGCDMDFDIAGPSIRGALVLGYEGWLAGYQMNFETAKSRVTQSNFAVGYKTDEFQLHTNVNDGTEFGGSIYQKVNKKLETAVNLAWTAGNSNTRFGIAAKYQIDPDACFSAKVNNSSLIGLGYTQTLKPGIKLTLSALLDGKNVNAGGHKLGLGLEFQA DIP protein P21796 original sequence version: 103 DIP protein P21796 original sequence version: 123 DIP protein P21796 original sequence version: 125 89BA3378B04020D5 psf2-1 3 6 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Fig. 5A Mass spectrometry analysis of the proteins identified the 36-kDa band as the mitochondrial ANT3, the 55-kDa band as beta tubulin, and the 80-kDa band as cytokeratin. The latter was likely a contaminant of the protein preparation. While interaction of PB1-F2 with tubulin may be important for PB1-F2-induced mitochondrial disorganization. pb1-slc25a6-1 4 6 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed physical association physical association Fig. 5B pb1-slc25a6-2 4 7 unspecified role unspecified role prey prey 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 n-term range n-terminal range n-term range n-term range n-terminal range n-term range 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed physical association physical association Fig. 5B pb1-vdac1-1 4 7 unspecified role unspecified role prey prey 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 n-term range n-terminal range n-term range n-term range n-terminal range n-term range 5 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Fig. 5E Immunoprecipitation experiments with anti-PB1-F2 polyclonal serum revealed that the PB1-F2 protein interacts with ANT3 and VDAC1 during the viral infection weak interaction pb1-vdac1-2 4 6 unspecified role unspecified role prey prey 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 n-term range n-terminal range n-term range n-term range n-terminal range n-term range 5 unspecified role unspecified role bait bait physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Fig. 5E Immunoprecipitation experiments with anti-PB1-F2 polyclonal serum revealed that the PB1-F2 protein interacts with ANT3 and VDAC1 during the viral infection pb1-slc25a6-3 1 6 unspecified role unspecified role prey prey cell lysate cell lysate in vitro translated in vitro translated protein in vitro translated 35s radiolabel 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes 5 unspecified role unspecified role bait bait purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Fig. 5D pb1-slc25a6-4 1 7 unspecified role unspecified role prey prey cell lysate cell lysate in vitro translated in vitro translated protein in vitro translated 35s radiolabel 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes 5 unspecified role unspecified role bait bait purified purified gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) physical association physical association Fig. 5D pb1-slc25a6-5 2 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range gfp tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminal_region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 6 unspecified role unspecified role prey prey 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 n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Fig. 6C pb1-slc25a6-6 2 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range gfp tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminal_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminal_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role prey prey 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 n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Fig. 6D