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/ http://www.ebi.ac.uk/ palty-2012-1 SARAF Inactivates the Store Operated Calcium Entry Machinery to Prevent Excess Calcium Refilling. 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. fluorescence imaging fluorescence microscopy fluorescence imaging tag fluorescence tag visualisation by fluorescence tag fluorescence Palty R., Raveh A., Kaminsky I., Meller R., Reuveny E. Source of STIM1 and ORA1 taken from previous work of contributing scientist (PMID: 16906149, 18995841) e.reuveny@weizmann.ac.il Cell (0092-8674) 2012 Only protein-protein interactions imex curation palty-2012-2 SARAF Inactivates the Store Operated Calcium Entry Machinery to Prevent Excess Calcium Refilling. 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 Palty R., Raveh A., Kaminsky I., Meller R., Reuveny E. Source of STIM1 and ORA1 taken from previous work of contributing scientist (PMID: 16906149, 18995841) e.reuveny@weizmann.ac.il Cell (0092-8674) 2012 Only protein-protein interactions imex curation palty-2012-3 SARAF Inactivates the Store Operated Calcium Entry Machinery to Prevent Excess Calcium Refilling. 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. fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Palty R., Raveh A., Kaminsky I., Meller R., Reuveny E. Source of STIM1 and ORA1 taken from previous work of contributing scientist (PMID: 16906149, 18995841) e.reuveny@weizmann.ac.il Cell (0092-8674) 2012 Only protein-protein interactions imex curation saraf_human Store-operated calcium entry-associated regulatory factor SARAF HBV X-transactivated gene 3 protein HBV XAg-transactivated protein 3 Protein FOAP-7 TMEM66 Transmembrane protein 66 XTP3 HSPC035 NPD003 PSEC0019 UNQ1967/PRO4499 protein protein human Homo sapiens Human MAAACGPGAAGYCLLLGLHLFLLTAGPALGWNDPDRMLLRDVKALTLHYDRYTTSRRLDPIPQLKCVGGTAGCDSYTPKVIQCQNKGWDGYDVQWECKTDLDIAYKFGKTVVSCEGYESSEDQYVLRGSCGLEYNLDYTELGLQKLKESGKQHGFASFSDYYYKWSSADSCNMSGLITIVVLLGIAFVVYKLFLSDGQYSPPPYSEYPPFSHRYQRFTNSAGPPPPGFKSEFTGPQNTGHGATSGFGSAFTGQQGYENSGPGFWTGLGTGGILGYLFGSNRAATPFSDSWYYPSYPPSYPGTWNRAYSPLHGGSGSYSVCSNSDTKTRTASGYGGTRRR stim1_human Stromal interaction molecule 1 STIM1 GOK protein protein human Homo sapiens Human MDVCVRLALWLLWGLLLHQGQSLSHSHSEKATGTSSGANSEESTAAEFCRIDKPLCHSEDEKLSFEAVRNIHKLMDDDANGDVDVEESDEFLREDLNYHDPTVKHSTFHGEDKLISVEDLWKAWKSSEVYNWTVDEVVQWLITYVELPQYEETFRKLQLSGHAMPRLAVTNTTMTGTVLKMTDRSHRQKLQLKALDTVLFGPPLLTRHNHLKDFMLVVSIVIGVGGCWFAYIQNRYSKEHMKKMMKDLEGLHRAEQSLHDLQERLHKAQEEHRTVEVEKVHLEKKLRDEINLAKQEAQRLKELREGTENERSRQKYAEEELEQVREALRKAEKELESHSSWYAPEALQKWLQLTHEVEVQYYNIKKQNAEKQLLVAKEGAEKIKKKRNTLFGTFHVAHSSSLDDVDHKILTAKQALSEVTAALRERLHRWQQIEILCGFQIVNNPGIHSLVAALNIDPSWMGSTRPNPAHFIMTDDVDDMDEEIVSPLSMQSPSLQSSVRQRLTEPQHGLGSQRDLTHSDSESSLHMSDRQRVAPKPPQMSRAADEALNAMTSNGSHRLIEGVHPGSLVEKLPDSPALAKKALLALNHGLDKAHSLMELSPSAPPGGSPHLDSSRSHSPSSPDPDTPSPVGDSRALQASRNTRIPHLAGKKAVAEEDNGSIGEETDSSPGRKKFPLKIFKKPLKK crcm1_human Calcium release-activated calcium channel protein 1 ORAI1 CRACM1 TMEM142A Protein orai-1 Transmembrane protein 142A protein protein human Homo sapiens Human MHPEPAPPPSRSSPELPPSGGSTTSGSRRSRRRSGDGEPPGAPPPPPSAVTYPDWIGQSYSEVMSLNEHSMQALSWRKLYLSRAKLKASSRTSALLSGFAMVAMVEVQLDADHDYPPGLLIAFSACTTVLVAVHLFALMISTCILPNIEAVSNVHNLNSVKESPHERMHRHIELAWAFSTVIGTLLFLAEVVLLCWVKFLPLKKQPGQPRPTSKPPASGAAANVSTSGITPGQAAAIASTTIMVPFGLIFIVFAVHFYRSLVSHKTDRQFQELNELAEFARLQDQLDHRGDHPLTPGSHYA stim1-tmem66 1291763 n/a 1291766 unspecified role unspecified role neutral component neutral component region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1291767 unspecified role unspecified role neutral component neutral component region mcherry mcherry fluorescent protein tag mcherry undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false Fig. 1G, 3B, Supp Fig. 5B and Movie S1 Addition of Thapsigargin (an irreversible sarcoplasmic/endoplasmic reticulum Ca2+-ATPase), induced translocation of the complex to the plasma membrane. orai1-tmem66 1291763 n/a 1291766 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1291768 unspecified role unspecified role neutral component neutral component region mcherry mcherry fluorescent protein tag mcherry undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false Fig. 3C, 3D, Supp. Fig. 4D Silencing STIM1 and STIM2 protein levels inhibited the Tg-induced SARAF-GFP translocation in the presence of Orai1-mCherry orai1-stim1-7 1291763 n/a 1291768 unspecified role unspecified role neutral component neutral component region mcherry mcherry fluorescent protein tag mcherry undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1291767 unspecified role unspecified role neutral component neutral component region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false Thapsigargin (an irreversible sarcoplasmic/endoplasmic reticulum Ca2+-ATPase) Fig. 6A tmem66-stim1 1291764 n/a 1291766 unspecified role unspecified role bait bait region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1291767 unspecified role unspecified role prey prey physical association physical association false false false Fig. 5C, 5D Traces of Orai1 immunopercipitate were also detected, which the authors attributed to its native interaction with STIM1 orai1-stim1-8 1291765 n/a 1291766 unspecified role unspecified role fluorescence donor fluorescence donor region yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1291767 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region mcherry mcherry fluorescent protein tag mcherry undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1291768 ancillary ancillary unspecified role unspecified role proximity proximity close-contact colocalization neighbourhood interaction false false false Fig. 4C, 5D, Supp. Fig. 6 results indicate that SARAF and STIM1 associate under resting conditions and that store operated calcium entry activation promotes molecular rearrangements between STIM1 and SARAF C termini in an Orai1-dependent manner.