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/ HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. Science (0036-8075) 2001 imex curation Ivan M., Kondo K., Yang H., Kim W., Valiando J., Ohh M., Salic A., Asara J.M., Lane W.S., Kaelin W.G. william_kaelin@dfci.harvard.edu 11-Jun-2014: Contacted by IntAct-Help. Only protein-protein interactions mouse-ts20 Mus musculus mouse-ts20 Thermosensitive E1 ubiquitin-activating enzyme embryonic fibroblast cell mutant A31N-ts20 anti bait coip anti bait coimmunoprecipitation anti bait coip autoradiography autoradiography Ivan M., Kondo K., Yang H., Kim W., Valiando J., Ohh M., Salic A., Asara J.M., Lane W.S., Kaelin W.G. ts20 cells were stably transfected to produce HA-pVHL were immunoprecipitated with anti–HIF-1a (lane 1 and 2) or anti-HA (lanes 3 to 7). Bound proteins were eluted by boiling in sample buffer (lane 1 and 2) or treatment with the indicated peptides, resolved by polyacrylamide gel electrophoresis, and detected by autoradiography. william_kaelin@dfci.harvard.edu Accepted 2014-MAY-27 by MGT Science (0036-8075) 2001 Only protein-protein interactions imex curation HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. Science (0036-8075) 2001 imex curation Ivan M., Kondo K., Yang H., Kim W., Valiando J., Ohh M., Salic A., Asara J.M., Lane W.S., Kaelin W.G. william_kaelin@dfci.harvard.edu 11-Jun-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting western blot western blot Immuno blot Immuno blot Ivan M., Kondo K., Yang H., Kim W., Valiando J., Ohh M., Salic A., Asara J.M., Lane W.S., Kaelin W.G. no Cloning, expression and purification of the recombinant VHL-elonginB-elonginC complex used as probe (detected by anti-VHL antibody) in the far western experiments is described in PMID:10205047. william_kaelin@dfci.harvard.edu Accepted 2014-MAY-27 by MGT Science (0036-8075) 2001 Only protein-protein interactions imex curation HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. Science (0036-8075) 2001 imex curation Ivan M., Kondo K., Yang H., Kim W., Valiando J., Ohh M., Salic A., Asara J.M., Lane W.S., Kaelin W.G. william_kaelin@dfci.harvard.edu 11-Jun-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down autoradiography autoradiography Ivan M., Kondo K., Yang H., Kim W., Valiando J., Ohh M., Salic A., Asara J.M., Lane W.S., Kaelin W.G. william_kaelin@dfci.harvard.edu Accepted 2014-MAY-27 by MGT Science (0036-8075) 2001 Only protein-protein interactions imex curation HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. Science (0036-8075) 2001 imex curation Ivan M., Kondo K., Yang H., Kim W., Valiando J., Ohh M., Salic A., Asara J.M., Lane W.S., Kaelin W.G. william_kaelin@dfci.harvard.edu 11-Jun-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down autoradiography autoradiography Ivan M., Kondo K., Yang H., Kim W., Valiando J., Ohh M., Salic A., Asara J.M., Lane W.S., Kaelin W.G. no Cloning, expression and purification of the GST-tagged (on VHL) recombinant VHL-elonginB-elonginC complex used as bait is described in PMID:10205047. The construct containing Gal4 DNA-binding domain, HA tag and Hif1-alpha is described in PMID:10878807. william_kaelin@dfci.harvard.edu Accepted 2014-MAY-27 by MGT Science (0036-8075) 2001 Only protein-protein interactions imex curation vhl_human von Hippel-Lindau disease tumor suppressor VHL pVHL Protein G7 protein protein human Homo sapiens Human MPRRAENWDEAEVGAEEAGVEEYGPEEDGGEESGAEESGPEESGPEELGAEEEMEAGRPRPVLRSVNSREPSQVIFCNRSPRVVLPVWLNFDGEPQPYPTLPPGTGRRIHSYRGHLWLFRDAGTHDGLLVNQTELFVPSLNVDGQPIFANITLPVYTLKERCLQVVRSLVKPENYRRLDIVRSLYEDLEDHPNVQKDLERLTQERIAHQRMGD DIP protein P40337 original sequence version: 176 DIP protein P40337 original sequence version: 103 DIP protein P40337-1 original sequence version: 177 DIP protein P40337 original sequence version: 160 DIP protein P40337 original sequence version: 175 BA5D6765FBC16EA7 hif1a_human Hypoxia-inducible factor 1-alpha ARNT-interacting protein Member of PAS protein 1 Basic-helix-loop-helix-PAS protein MOP1 HIF1A MOP1 BHLHE78 PASD8 PAS domain-containing protein 8 Class E basic helix-loop-helix protein 78 protein protein human Homo sapiens Human MEGAGGANDKKKISSERRKEKSRDAARSRRSKESEVFYELAHQLPLPHNVSSHLDKASVMRLTISYLRVRKLLDAGDLDIEDDMKAQMNCFYLKALDGFVMVLTDDGDMIYISDNVNKYMGLTQFELTGHSVFDFTHPCDHEEMREMLTHRNGLVKKGKEQNTQRSFFLRMKCTLTSRGRTMNIKSATWKVLHCTGHIHVYDTNSNQPQCGYKKPPMTCLVLICEPIPHPSNIEIPLDSKTFLSRHSLDMKFSYCDERITELMGYEPEELLGRSIYEYYHALDSDHLTKTHHDMFTKGQVTTGQYRMLAKRGGYVWVETQATVIYNTKNSQPQCIVCVNYVVSGIIQHDLIFSLQQTECVLKPVESSDMKMTQLFTKVESEDTSSLFDKLKKEPDALTLLAPAAGDTIISLDFGSNDTETDDQQLEEVPLYNDVMLPSPNEKLQNINLAMSPLPTAETPKPLRSSADPALNQEVALKLEPNPESLELSFTMPQIQDQTPSPSDGSTRQSSPEPNSPSEYCFYVDSDMVNEFKLELVEKLFAEDTEAKNPFSTQDTDLDLEMLAPYIPMDDDFQLRSFDQLSPLESSSASPESASPQSTVTVFQQTQIQEPTANATTTTATTDELKTVTKDRMEDIKILIASPSPTHIHKETTSATSSPYRDTQSRTASPNRAGKGVIEQTEKSHPRSPNVLSVALSQRTTVPEEELNPKILALQNAQRKRKMEHDGSLFQAVGIGTLLQQPDDHAATTSLSWKRVKGCKSSEQNGMEQKTIILIPSDLACRLLGQSMDESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRALDQVN DIP protein Q16665 original sequence version: 94 DIP protein Q16665 original sequence version: 98 DIP protein Q16665-1 original sequence version: 102 DIP protein Q16665 original sequence version: 102 DIP protein Q16665 original sequence version: 127 DIP protein Q16665 original sequence version: 120 DIP protein Q16665 original sequence version: 169 DIP protein Q16665 original sequence version: 174 DIP protein Q16665-1 original sequence version: 174 DIP protein Q16665-1 original sequence version: 191 DIP protein Q16665 original sequence version: 191 DIP protein Q16665 original sequence version: 179 DIP protein Q16665 original sequence version: 150 DIP protein Q16665 original sequence version: 156 DIP protein Q16665 original sequence version: 177 ABD4F7DAA135BE2D epas1_human Endothelial PAS domain-containing protein 1 EPAS1 HIF2A MOP2 Member of PAS protein 2 Basic-helix-loop-helix-PAS protein MOP2 BHLHE73 PASD2 Hypoxia-inducible factor 2-alpha HIF-1-alpha-like factor PAS domain-containing protein 2 Class E basic helix-loop-helix protein 73 protein protein human Homo sapiens Human MTADKEKKRSSSERRKEKSRDAARCRRSKETEVFYELAHELPLPHSVSSHLDKASIMRLAISFLRTHKLLSSVCSENESEAEADQQMDNLYLKALEGFIAVVTQDGDMIFLSENISKFMGLTQVELTGHSIFDFTHPCDHEEIRENLSLKNGSGFGKKSKDMSTERDFFMRMKCTVTNRGRTVNLKSATWKVLHCTGQVKVYNNCPPHNSLCGYKEPLLSCLIIMCEPIQHPSHMDIPLDSKTFLSRHSMDMKFTYCDDRITELIGYHPEELLGRSAYEFYHALDSENMTKSHQNLCTKGQVVSGQYRMLAKHGGYVWLETQGTVIYNPRNLQPQCIMCVNYVLSEIEKNDVVFSMDQTESLFKPHLMAMNSIFDSSGKGAVSEKSNFLFTKLKEEPEELAQLAPTPGDAIISLDFGNQNFEESSAYGKAILPPSQPWATELRSHSTQSEAGSLPAFTVPQAAAPGSTTPSATSSSSSCSTPNSPEDYYTSLDNDLKIEVIEKLFAMDTEAKDQCSTQTDFNELDLETLAPYIPMDGEDFQLSPICPEERLLAENPQSTPQHCFSAMTNIFQPLAPVAPHSPFLLDKFQQQLESKKTEPEHRPMSSIFFDAGSKASLPPCCGQASTPLSSMGGRSNTQWPPDPPLHFGPTKWAVGDQRTEFLGAAPLGPPVSPPHVSTFKTRSAKGFGARGPDVLSPAMVALSNKLKLKRQLEYEEQAFQDLSGGDPPGGSTSHLMWKRMKNLRGGSCPLMPDKPLSANVPNDKFTQNPMRGLGHPLRHLPLPQPPSAISPGENSKSRFPPQCYATQYQDYSLSSAHKVSGMASRLLGPSFESYLLPELTRYDCEVNVPVLGSSTLLQGGDLLRALDQAT DIP protein Q99814 original sequence version: 115 DIP protein Q99814 original sequence version: 155 DIP protein Q99814 original sequence version: 157 DIP protein Q99814 original sequence version: 137 DIP protein Q99814 original sequence version: 135 4838989598234FC1 hif1a_mouse Hypoxia-inducible factor 1-alpha Hif1a ARNT-interacting protein protein protein mouse Mus musculus Mouse MEGAGGENEKKKMSSERRKEKSRDAARSRRSKESEVFYELAHQLPLPHNVSSHLDKASVMRLTISYLRVRKLLDAGGLDSEDEMKAQMDCFYLKALDGFVMVLTDDGDMVYISDNVNKYMGLTQFELTGHSVFDFTHPCDHEEMREMLTHRNGPVRKGKELNTQRSFFLRMKCTLTSRGRTMNIKSATWKVLHCTGHIHVYDTNSNQPQCGYKKPPMTCLVLICEPIPHPSNIEIPLDSKTFLSRHSLDMKFSYCDERITELMGYEPEELLGRSIYEYYHALDSDHLTKTHHDMFTKGQVTTGQYRMLAKRGGYVWVETQATVIYNTKNSQPQCIVCVNYVVSGIIQHDLIFSLQQTESVLKPVESSDMKMTQLFTKVESEDTSCLFDKLKKEPDALTLLAPAAGDTIISLDFGSDDTETEDQQLEDVPLYNDVMFPSSNEKLNINLAMSPLPSSETPKPLRSSADPALNQEVALKLESSPESLGLSFTMPQIQDQPASPSDGSTRQSSPERLLQENVNTPNFSQPNSPSEYCFDVDSDMVNVFKLELVEKLFAEDTEAKNPFSTQDTDLDLEMLAPYIPMDDDFQLRSFDQLSPLESNSPSPPSMSTVTGFQQTQLQKPTITATATTTATTDESKTETKDNKEDIKILIASPSSTQVPQETTTAKASAYSGTHSRTASPDRAGKRVIEQTDKAHPRSLNLSATLNQRNTVPEEELNPKTIASQNAQRKRKMEHDGSLFQAAGIGTLLQQPGDCAPTMSLSWKRVKGFISSEQNGTEQKTIILIPSDLACRLLGQSMDESGLPQLTSYDCEVNAPIQGSRNLLQGEELLRALDQVN 84D64ECAC2CC753B hif1a-vhl-5 3 5 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 6 unspecified role unspecified role bait bait deg_odph_vhl_1 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain required hydroxypro-564 4hypro 4-hydroxy-L-proline (2S,4R)-4-hydroxypyrrolidine-2-carboxylic acid 4-hydroxy-L-proline 4-hydroxylated L-proline 4-hydroxyproline 4-trans-hydroxy-L-proline L-threo-4-hydroxyproline MOD_RES 4-hydroxyproline gamma-hydroxypyrrolidine-alpha-carboxylic acid 4HyPro modified residue ms mass detection of residue modification modified residue ms certain certain sequence position certain certain certain sequence position certain hydroxylation of pro564 region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q16665:p.Pro564Ala mutation disrupting mutation disrupting interaction mutation disrupting alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain P A Wed May 02 16:03:13 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro564ala' Tue Jun 30 17:19:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro564Ala' Q16665:p.Leu562Ala mutation disrupting mutation disrupting interaction mutation disrupting alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain L A Wed May 02 17:15:31 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu562ala' Tue Jun 30 17:28:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu562Ala' Q16665:p.[Met561_Leu562delinsAlaAla;Pro564_Met568delinsAlaAlaAlaAlaAla] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain ML AA certain certain sequence position certain certain certain sequence position certain PYIPM AAAAA Thu Jul 07 12:17:15 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was met561-leu562-pro564-tyr565-ile566-pro567-met568ala Fri Jul 22 11:03:10 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'met561ala,leu562ala,pro564ala,tyr565ala,ile566ala,pro567ala,met568ala' Wed May 02 14:34:45 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'met_leu561-562ala_ala,pro_tyr_ile_pro_met564-568ala_ala_ala_ala_ala' Tue Jun 30 15:48:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Met561_Leu562delinsAlaAla;Pro564_Met568delinsAlaAlaAlaAlaAla]' chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association Fig. 2b, Fig. 4, Fig 5a, Fig. 5b The interaction only occurs after chemically synthesized Hif1-alpha peptides are incubated with rabbit reticulocyte lysate. vhl-epas1-1 2 5 unspecified role unspecified role bait bait vhl_tumour_suppress_b/a_dom sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobd Escherichia coli (strain B / BL21-DE3) 7 unspecified role unspecified role prey prey human-786_o Homo sapiens renal cell adenocarcinoma 786_o Human renal cell adenocarcinoma cell line CRL-1932 is the ATCC identifier. ATCC identifier is CRL-1932. association association Fig. 1a Increasing amounts of cobalt chloride (CoCl2) (2, 10, 100, 1000 micromolar) or desferrioxamine (DFO) (2, 10, 100, 1000 micromolar) inhibits the interaction. vhl-hif1a-6 2 8 unspecified role unspecified role prey prey mouse-ts20 Mus musculus mouse-ts20 Thermosensitive E1 ubiquitin-activating enzyme embryonic fibroblast cell mutant A31N-ts20 5 unspecified role unspecified role bait bait vhl_tumour_suppress_b/a_dom sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobd Escherichia coli (strain B / BL21-DE3) association association Fig. 1b Hypoxic condition (1% oxygen) inhibits the interaction. vhl-hif1a-7 2 5 unspecified role unspecified role bait bait vhl_tumour_suppress_b/a_dom sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobd Escherichia coli (strain B / BL21-DE3) 6 unspecified role unspecified role prey prey region 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobd Escherichia coli (strain B / BL21-DE3) association association Fig. 1c, Fig. 1d The interaction only occurs after bacterially expressed Hif1-alpha is incubated with rabbit reticulocyte lysate (for 90 minutes at 30C). Heat-inactivated rabbit reticulocyte lysate does not have this effect. vhl-hif1a-8 4 5 unspecified role unspecified role bait bait vhl_tumour_suppress_b/a_dom sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n-terminus 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 ecobd Escherichia coli (strain B / BL21-DE3) 6 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region gal4 dna bd gal4 dna binding domain gal4 dna bd undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q16665:p.Leu562Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L A Wed May 02 17:26:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu562ala' Tue Jun 30 17:52:28 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu562Ala' Q16665:p.Pro564Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain P A Wed May 02 20:09:09 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro564ala' Tue Jun 30 19:53:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro564Ala' region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro physical association physical association Fig. 2a, Fig. 2c, Fig. 2d cul2-hif1a 3 6 unspecified role unspecified role bait bait deg_odph_vhl_1 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q16665:p.Pro564Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain P A Wed May 02 17:40:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro564ala' Tue Jun 30 18:32:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro564Ala' required hydroxypro-564 4hypro 4-hydroxy-L-proline (2S,4R)-4-hydroxypyrrolidine-2-carboxylic acid 4-hydroxy-L-proline 4-hydroxylated L-proline 4-hydroxyproline 4-trans-hydroxy-L-proline L-threo-4-hydroxyproline MOD_RES 4-hydroxyproline gamma-hydroxypyrrolidine-alpha-carboxylic acid 4HyPro certain certain sequence position certain certain certain sequence position certain hydroxylation of pro564 chemical synthesis Chemical synthesis (Chemical synthesis) 5 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined human-786_o Homo sapiens renal cell adenocarcinoma 786_o Human renal cell adenocarcinoma cell line CRL-1932 is the ATCC identifier. ATCC identifier is CRL-1932. physical association physical association Fig. 5d The hydroxylated HIF(556-575) peptide specifically bound to pVHL as well as to proteins with the expected electrophoretic mobilities of the pVHL-associated proteins elongin B, elongin C, and Cul2. vhl-hif1a-9 1 6 competitor competitor competitor prey prey deg_odph_vhl_1 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q16665:p.Pro564Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain P A Wed May 02 15:53:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro564ala' Tue Jun 30 17:47:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro564Ala' required hydroxypro-564 4hypro 4-hydroxy-L-proline (2S,4R)-4-hydroxypyrrolidine-2-carboxylic acid 4-hydroxy-L-proline 4-hydroxylated L-proline 4-hydroxyproline 4-trans-hydroxy-L-proline L-threo-4-hydroxyproline MOD_RES 4-hydroxyproline gamma-hydroxypyrrolidine-alpha-carboxylic acid 4HyPro certain certain sequence position certain certain certain sequence position certain hydroxylation of pro564 8 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 5c vhl-hif1a-10 1 5 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey physical association physical association Fig. 5ci