UniProt UniProt http://www.uniprot.org Siva-1 binds to and inhibits BCL-X(L)-mediated protection against UV radiation-induced apoptosis. Siva-1 binds to and inhibits BCL-X(L)-mediated protection against UV radiation-induced apoptosis. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 Xue L., Chu F., Cheng Y., Sun X., Borthakur A., Ramarao M., Pandey P., Wu M., Schlossman SF., Prasad KV. Apoptosis - Interactions involving proteins with a function related to apoptosis Identity of BCL-XL assumed from previous work of contributer. Kanteti@uic.edu 12-Oct-2007: Contacted by JYOTI. orchard human-hut78 Homo sapiens HUT-78 T cell lymphoma hut78 HUT-78 human T cell lymphoma anti bait coip anti bait coimmunoprecipitation anti bait coip antibody detection identification by antibody antibody detection Xue L., Chu F., Cheng Y., Sun X., Borthakur A., Ramarao M., Pandey P., Wu M., Schlossman SF., Prasad KV. Apoptosis - Interactions involving proteins with a function related to apoptosis Identity of BCL-XL assumed from previous work of contributer. Kanteti@uic.edu Accepted 2007-OCT-21 by ORCHARD. 12-Oct-2007: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 orchard Siva-1 binds to and inhibits BCL-X(L)-mediated protection against UV radiation-induced apoptosis. Siva-1 binds to and inhibits BCL-X(L)-mediated protection against UV radiation-induced apoptosis. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 Xue L., Chu F., Cheng Y., Sun X., Borthakur A., Ramarao M., Pandey P., Wu M., Schlossman SF., Prasad KV. Apoptosis - Interactions involving proteins with a function related to apoptosis Identity of BCL-XL assumed from previous work of contributer. Kanteti@uic.edu 12-Oct-2007: Contacted by JYOTI. orchard chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 pull down pull down anti tag western anti tag western blot anti tag western Xue L., Chu F., Cheng Y., Sun X., Borthakur A., Ramarao M., Pandey P., Wu M., Schlossman SF., Prasad KV. Apoptosis - Interactions involving proteins with a function related to apoptosis Identity of BCL-XL assumed from previous work of contributer. Kanteti@uic.edu Accepted 2007-OCT-21 by ORCHARD. 12-Oct-2007: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 orchard Siva-1 binds to and inhibits BCL-X(L)-mediated protection against UV radiation-induced apoptosis. Siva-1 binds to and inhibits BCL-X(L)-mediated protection against UV radiation-induced apoptosis. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 Xue L., Chu F., Cheng Y., Sun X., Borthakur A., Ramarao M., Pandey P., Wu M., Schlossman SF., Prasad KV. Apoptosis - Interactions involving proteins with a function related to apoptosis Identity of BCL-XL assumed from previous work of contributer. Kanteti@uic.edu 12-Oct-2007: Contacted by JYOTI. orchard mouse-thymus Mus musculus thymus thymus anti bait coip anti bait coimmunoprecipitation anti bait coip antibody detection identification by antibody antibody detection Xue L., Chu F., Cheng Y., Sun X., Borthakur A., Ramarao M., Pandey P., Wu M., Schlossman SF., Prasad KV. Apoptosis - Interactions involving proteins with a function related to apoptosis Identity of BCL-XL assumed from previous work of contributer. Kanteti@uic.edu Accepted 2007-OCT-21 by ORCHARD. 12-Oct-2007: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 orchard Siva-1 binds to and inhibits BCL-X(L)-mediated protection against UV radiation-induced apoptosis. Siva-1 binds to and inhibits BCL-X(L)-mediated protection against UV radiation-induced apoptosis. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 Xue L., Chu F., Cheng Y., Sun X., Borthakur A., Ramarao M., Pandey P., Wu M., Schlossman SF., Prasad KV. Apoptosis - Interactions involving proteins with a function related to apoptosis Identity of BCL-XL assumed from previous work of contributer. Kanteti@uic.edu 12-Oct-2007: Contacted by JYOTI. orchard in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Xue L., Chu F., Cheng Y., Sun X., Borthakur A., Ramarao M., Pandey P., Wu M., Schlossman SF., Prasad KV. Apoptosis - Interactions involving proteins with a function related to apoptosis Identity of BCL-XL assumed from previous work of contributer. Kanteti@uic.edu Accepted 2007-OCT-21 by ORCHARD. 12-Oct-2007: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 orchard Siva-1 binds to and inhibits BCL-X(L)-mediated protection against UV radiation-induced apoptosis. Siva-1 binds to and inhibits BCL-X(L)-mediated protection against UV radiation-induced apoptosis. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 Xue L., Chu F., Cheng Y., Sun X., Borthakur A., Ramarao M., Pandey P., Wu M., Schlossman SF., Prasad KV. Apoptosis - Interactions involving proteins with a function related to apoptosis Identity of BCL-XL assumed from previous work of contributer. Kanteti@uic.edu 12-Oct-2007: Contacted by JYOTI. orchard in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Xue L., Chu F., Cheng Y., Sun X., Borthakur A., Ramarao M., Pandey P., Wu M., Schlossman SF., Prasad KV. Apoptosis - Interactions involving proteins with a function related to apoptosis Identity of BCL-XL assumed from previous work of contributer. Kanteti@uic.edu Accepted 2007-OCT-21 by ORCHARD. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 orchard Siva-1 binds to and inhibits BCL-X(L)-mediated protection against UV radiation-induced apoptosis. Siva-1 binds to and inhibits BCL-X(L)-mediated protection against UV radiation-induced apoptosis. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 Xue L., Chu F., Cheng Y., Sun X., Borthakur A., Ramarao M., Pandey P., Wu M., Schlossman SF., Prasad KV. Apoptosis - Interactions involving proteins with a function related to apoptosis Identity of BCL-XL assumed from previous work of contributer. Kanteti@uic.edu 12-Oct-2007: Contacted by JYOTI. orchard human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 confocal microscopy confocal microscopy anti tag immunost anti tag immunostaining anti tag immunost Xue L., Chu F., Cheng Y., Sun X., Borthakur A., Ramarao M., Pandey P., Wu M., Schlossman SF., Prasad KV. Apoptosis - Interactions involving proteins with a function related to apoptosis Identity of BCL-XL assumed from previous work of contributer. Kanteti@uic.edu Accepted 2007-OCT-21 by ORCHARD. 12-Oct-2007: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2002 orchard siva_mouse Apoptosis regulatory protein Siva Siva1 Siva CD27-binding protein protein protein mouse Mus musculus Mouse MPKRSCPFADAAPLQLKVHVGLKELSHGVFAERYSREVFERTKQLLFQGARAYRDHISSEDCSVNHLQESLKSGVVGAPQPARGQMLIGPDGRLTRCQAQASEGGLPRTAPIACSSCMRSVDGKAVCSQCERALCGQCVYTCWGCGALACVLCGLADYADDGEKTLCTSCAMFEA 1E132C1580ABE3F2 q07817-1 Bcl-2-like protein 1 BCLX Apoptosis regulator Bcl-X BCL2L1 BCL2L Bcl-xL protein protein human Homo sapiens Human MSQSNRELVVDFLSYKLSQKGYSWSQFSDVEENRTEAPEGTESEMETPSAINGNPSWHLADSPAVNGATGHSSSLDAREVIPMAAVKQALREAGDEFELRYRRAFSDLTSQLHITPGTAYQSFEQVVNELFRDGVNWGRIVAFFSFGGALCVESVDKEMQVLVSRIAAWMATYLNDHLEPWIQENGGWDTFVELYGNNAAAESRKGQERFNRWFLTGMTVAGVVLLGSLFSRK DIP protein Q07817 original sequence version: 82 DIP protein Q07817 original sequence version: 86 DIP protein Q07817-1 original sequence version: 100 DIP protein Q07817-1 original sequence version: 108 DIP protein Q07817 original sequence version: 131 DIP protein Q07817-1 original sequence version: 110 DIP protein Q07817-1 original sequence version: 135 DIP protein Q07817-1 original sequence version: 131 DIP protein Q07817-1 original sequence version: 172 DIP protein Q07817-1 original sequence version: 174 DIP protein Q07817 original sequence version: 174 DIP protein Q07817-1 original sequence version: 189 DIP protein Q07817-1 original sequence version: 192 DIP protein Q07817 original sequence version: 190 E09D3CDD851AE9BE q64373-1 Bcl-2-like protein 1 Bcl2l1 Bcl2l Bclx Apoptosis regulator Bcl-X Bcl-xL protein protein mouse Mus musculus Mouse MSQSNRELVVDFLSYKLSQKGYSWSQFSDVEENRTEAPEETEAERETPSAINGNPSWHLADSPAVNGATGHSSSLDAREVIPMAAVKQALREAGDEFELRYRRAFSDLTSQLHITPGTAYQSFEQVVNELFRDGVNWGRIVAFFSFGGALCVESVDKEMQVLVSRIASWMATYLNDHLEPWIQENGGWDTFVDLYGNNAAAESRKGQERFNRWFLTGMTVAGVVLLGSLFSRK 24D2AC79887E072E o15304-1 Apoptosis regulatory protein Siva Siva-1 SIVA1 SIVA CD27-binding protein protein protein human Homo sapiens Human MPKRSCPFADVAPLQLKVRVSQRELSRGVCAERYSQEVFEKTKRLLFLGAQAYLDHVWDEGCAVVHLPESPKPGPTGAPRAARGQMLIGPDGRLIRSLGQASEADPSGVASIACSSCVRAVDGKAVCGQCERALCGQCVRTCWGCGSVACTLCGLVDCSDMYEKVLCTSCAMFET DB62B7DF4E6B39B3 siva_human Apoptosis regulatory protein Siva CD27-binding protein SIVA1 SIVA protein protein human Homo sapiens Human MPKRSCPFADVAPLQLKVRVSQRELSRGVCAERYSQEVFEKTKRLLFLGAQAYLDHVWDEGCAVVHLPESPKPGPTGAPRAARGQMLIGPDGRLIRSLGQASEADPSGVASIACSSCVRAVDGKAVCGQCERALCGQCVRTCWGCGSVACTLCGLVDCSDMYEKVLCTSCAMFET DB62B7DF4E6B39B3 siva1-bcl2l1-3 5 8 unspecified role unspecified role prey prey region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role bait bait 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction 2A Interaction was dose-dependent on BCL2L1. GST-Siva-1 but not GST alone coprecipitated BCL-XL with increasing amounts of soluble BCL-XL, demonstrating direct interaction between Siva-1 and BCL-XL. siva1-bcl2l1-5 5 10 unspecified role unspecified role bait bait 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 ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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. physical association physical association 2B Also, GST-Siva-1 and GST-BCL-XL proteins were used to pull down GFP-BCL-XL and GFP-Siva-1, respectively, from 293T cell lysates (Fig. 2B). siva1-bcl2l1-6 2 8 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role bait bait 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 region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association 2C and 4A Immunoblotting with anti-GFP antibodies revealed that GST-Siva-1 but not GST-Siva-2 or GST is able to coprecipitate GFP-BCL-XL from the double transfectants, and GFP is unable to associate with any of the GST fusion proteins (Fig. 2C; siva1-bcl2l1-7 6 8 unspecified role unspecified role neutral component neutral component region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization 3A The two patterns are highly superimposable (yellow) with punctate distribution around the nucleus characteristic of mitochondrial expression, favoring Siva-1/BCL-XL interaction in the cell. bcl2l1-siva1-3 3 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait physical association physical association 3B The kinetics of natural Siva-1/BCL-XL complexes in murine thymocytes after CD3 crosslinking is shown. Amount of Siva bound to Bcl-XL increased on CD3 cross-linking siva1-bcl2l1-8 1 8 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association 3C Additionally, we detected a significant presence of BCL-XL in anti-Siva immunoprecipitates but not in other control immunoprecipitations (Lower). siva1-bcl2l1-10 4 8 unspecified role unspecified role bait bait 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 ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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. physical association physical association 2B Also, GST-Siva-1 and GST-BCL-XL proteins were used to pull down GFP-BCL-XL and GFP-Siva-1, respectively, from 293T cell lysates (Fig. 2B). siva1-bcl2l1-9 1 11 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association 3C With untreated HUT78 cells (Fig. 3C), we observed that a significant amount of Siva-1 coprecipitated with BCL-XL but not with BCL-2 or with the secondary antibody-conjugated beads (Upper).