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/ SOS2 promotes salt tolerance in part by interacting with the vacuolar H+-ATPase and upregulating its transport activity. SOS2 promotes salt tolerance in part by interacting with the vacuolar H+-ATPase and upregulating its transport activity. Mol. Cell. Biol. (0270-7306) 2007 imex curation Batelli G., Verslues PE., Agius F., Qiu Q., Fujii H., Pan S., Schumaker KS., Grillo S., Zhu JK. Note the TAP tagging assay with detection of proteins using western blot could not be curated as the authors fail to specifity of the antibody used to detect VHA-B (isoforms of VHA-B are given as HA-B1, VHA-B2 or VHA-B3). Arabidopsis plants (SOS2 deficient mutants) were transformed with a C-terminal tagged SOS2 protein expressing construct. Twenty day old transgenic plants were exposed to 150 millimolar Sodium Chloride solution for 24 hours and then the TAP purification protocol carried out. jian-kang.zhu@ucr.edu 03-Feb-2009: Contacted by JYOTI. orchard Only protein-protein interactions arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress tap tandem affinity purification TAP TAP tap ms participant Identification by mass spectrometry ms participant Batelli G., Verslues PE., Agius F., Qiu Q., Fujii H., Pan S., Schumaker KS., Grillo S., Zhu JK. Note the TAP tagging assay with detection of proteins using western blot could not be curated as the authors fail to specifity of the antibody used to detect VHA-B (isoforms of VHA-B are given as HA-B1, VHA-B2 or VHA-B3). Arabidopsis plants (SOS2 deficient mutants) were transformed with a C-terminal tagged SOS2 protein expressing construct. Twenty day old transgenic plants were exposed to 150 millimolar Sodium Chloride solution for 24 hours and then the TAP purification protocol carried out. jian-kang.zhu@ucr.edu Accepted 2009-FEB-19 by ORCHARD. 03-Mar-2009: Contacted by JYOTI. Mol. Cell. Biol. (0270-7306) 2007 orchard Only protein-protein interactions imex curation SOS2 promotes salt tolerance in part by interacting with the vacuolar H+-ATPase and upregulating its transport activity. SOS2 promotes salt tolerance in part by interacting with the vacuolar H+-ATPase and upregulating its transport activity. Mol. Cell. Biol. (0270-7306) 2007 imex curation Batelli G., Verslues PE., Agius F., Qiu Q., Fujii H., Pan S., Schumaker KS., Grillo S., Zhu JK. Note the TAP tagging assay with detection of proteins using western blot could not be curated as the authors fail to specifity of the antibody used to detect VHA-B (isoforms of VHA-B are given as HA-B1, VHA-B2 or VHA-B3). Arabidopsis plants (SOS2 deficient mutants) were transformed with a C-terminal tagged SOS2 protein expressing construct. Twenty day old transgenic plants were exposed to 150 millimolar Sodium Chloride solution for 24 hours and then the TAP purification protocol carried out. jian-kang.zhu@ucr.edu 03-Feb-2009: Contacted by JYOTI. orchard Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Batelli G., Verslues PE., Agius F., Qiu Q., Fujii H., Pan S., Schumaker KS., Grillo S., Zhu JK. Note the TAP tagging assay with detection of proteins using western blot could not be curated as the authors fail to specifity of the antibody used to detect VHA-B (isoforms of VHA-B are given as HA-B1, VHA-B2 or VHA-B3). Arabidopsis plants (SOS2 deficient mutants) were transformed with a C-terminal tagged SOS2 protein expressing construct. Twenty day old transgenic plants were exposed to 150 millimolar Sodium Chloride solution for 24 hours and then the TAP purification protocol carried out. jian-kang.zhu@ucr.edu Accepted 2009-FEB-19 by ORCHARD. 03-Feb-2009: Contacted by JYOTI. Mol. Cell. Biol. (0270-7306) 2007 orchard Only protein-protein interactions imex curation vatb3_arath V-type proton ATPase subunit B3 VHA-B3 Vacuolar proton pump subunit B3 Vacuolar H(+)-ATPase subunit B isoform 3 F14O10.13 At1g20260 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MVETSIDMEEGTLEIGMEYRTVSGVAGPLVILDKVKGPKYQEIVNIRLGDGSTRRGQVLEVDGEKAVVQVFEGTSGIDNKFTTVQFTGEVLKTPVSLDMLGRIFNGSGKPIDNGPPILPEAYLDISGSSINPSERTYPEEMIQTGISTIDVMNSIARGQKIPLFSAAGLPHNEIAAQICRQAGLVKRLEKTENLIQEDHGEDNFAIVFAAMGVNMETAQFFKRDFEENGSMERVTLFLNLANDPTIERIITPRIALTTAEYLAYECGKHVLVILTDMSSYADALREVSAAREEVPGRRGYPGYMYTDLATIYERAGRIEGRKGSITQIPILTMPNDDITHPTPDLTGYITEGQIYIDRQLHNRQIYPPINVLPSLSRLMKSAIGEGMTRKDHSDVSNQLYANYAIGKDVQAMKAVVGEEALSSEDLLYLEFLDKFERKFVMQGAYDTRNIFQSLDLAWTLLRIFPRELLHRIPAKTLDQFYSRDSTS B833A2CAE4D42CE8 vatb2_arath V-type proton ATPase subunit B2 VHA-B2 F20M13.70 At4g38510 Vacuolar proton pump subunit B2 Vacuolar H(+)-ATPase subunit B isoform 2 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MGAAENNLEMEGTLEIGMEYRTVSGVAGPLVILEKVKGPKYQEIVNIRLGDGTTRRGQVLEVDGEKAVVQVFEGTSGIDNKYTTVQFTGEVLKTPVSLDMLGRIFNGSGKPIDNGPPILPEAYLDISGSSINPSERTYPEEMIQTGISTIDVMNSIARGQKIPLFSAAGLPHNEIAAQICRQAGLVKRLEKSDNLLEHQEDDNFAIVFAAMGVNMETAQFFKRDFEENGSMERVTLFLNLANDPTIERIITPRIALTTAEYLAYECGKHVLVILTDMSSYADALREVSAAREEVPGRRGYPGYMYTDLATIYERAGRIEGRKGSITQIPILTMPNDDITHPTPDLTGYITEGQIYIDRQLHNRQIYPPINVLPSLSRLMKSAIGEGMTRRDHSDVSNQLYANYAIGKDVQAMKAVVGEEALSSEDLLYLEFLDKFERKFVAQGAYDTRNIFQSLDLAWTLLRIFPRELLHRIPAKTLDQFYSRDTTN 170224B66A22F1F1 vatg1_arath V-type proton ATPase subunit G1 VATG1 AVMA10 VAG1 VMA10 T13O15.3 At3g01390 VHA-G1 Vacuolar proton pump subunit G1 Vacuolar H(+)-ATPase subunit G isoform 1 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MESNRGQGSIQQLLAAEVEAQHIVNAARTAKMARLKQAKEEAEKEIAEYKAQTEQDFQRKLEETSGDSGANVKRLEQETDTKIEQLKNEASRISKDVVEMLLKHVTTVKN C486D697AD434142 cipko_arath CBL-interacting serine/threonine-protein kinase 24 SNF1-related kinase 3.11 Protein SALT OVERLY SENSITIVE 2 CIPK24 SnRK3.11 SOS2 At5g35410 F6I13.1 K21B8.3 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MTKKMRRVGKYEVGRTIGEGTFAKVKFARNTDTGDNVAIKIMAKSTILKNRMVDQIKREISIMKIVRHPNIVRLYEVLASPSKIYIVLEFVTGGELFDRIVHKGRLEESESRKYFQQLVDAVAHCHCKGVYHRDLKPENLLLDTNGNLKVSDFGLSALPQEGVELLRTTCGTPNYVAPEVLSGQGYDGSAADIWSCGVILFVILAGYLPFSETDLPGLYRKINAAEFSCPPWFSAEVKFLIHRILDPNPKTRIQIQGIKKDPWFRLNYVPIRAREEEEVNLDDIRAVFDGIEGSYVAENVERNDEGPLMMNAFEMITLSQGLNLSALFDRRQDFVKRQTRFVSRREPSEIIANIEAVANSMGFKSHTRNFKTRLEGLSSIKAGQLAVVIEIYEVAPSLFMVDVRKAAGETLEYHKFYKKLCSKLENIIWRATEGIPKSEILRTITF DIP protein Q9LDI3 original sequence version: 90 99FDF3A778E1093D vatb1_arath V-type proton ATPase subunit B1 T4O12.24 At1g76030 VHA-B1 V-ATPase 57 kDa subunit AT57 Vacuolar proton pump subunit B1 Vacuolar H(+)-ATPase subunit B isoform 1 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MGTNDLDIEEGTLEIGMEYRTVSGVAGPLVILDKVKGPKYQEIVNIRLGDGSTRRGQVLEVDGEKAVVQVFEGTSGIDNKFTTVQFTGEVLKTPVSLDMLGRIFNGSGKPIDNGPPILPEAYLDISGSSINPSERTYPEEMIQTGISTIDVMNSIARGQKIPLFSAAGLPHNEIAAQICRQAGLVKRLEKTVDLLEDHGEDNFAIVFAAMGVNMETAQFFKRDFEENGSMERVTLFLNLANDPTIERIITPRIALTTAEYLAYECGKHVLVILTDMSSYADALREVSAAREEVPGRRGYPGYMYTDLATIYERAGRIEGRKGSITQIPILTMPNDDITHPTPDLTGYITEGQIYIDRQLHNRQIYPPINVLPSLSRLMKSAIGEGMTRKDHSDVSNQLYANYAIGKDVQAMKAVVGEEALSSEDLLYLEFLDKFERKFVMQGAYDTRNIFQSLDLAWTLLRIFPRELLHRIPAKTLDQFYSRDSTS 0EC3B68E92CC9F0A vata_arath V-type proton ATPase catalytic subunit A At1g78900 F9K20.5 Vacuolar proton pump subunit alpha V-ATPase 69 kDa subunit VHA-A Vacuolar H(+)-ATPase subunit A protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MPAFYGGKLTTFEDDEKESEYGYVRKVSGPVVVADGMAGAAMYELVRVGHDNLIGEIIRLEGDSATIQVYEETAGLTVNDPVLRTHKPLSVELGPGILGNIFDGIQRPLKTIARISGDVYIPRGVSVPALDKDCLWEFQPNKFVEGDTITGGDLYATVFENTLMNHLVALPPDAMGKITYIAPAGQYSLKDTVIELEFQGIKKSYTMLQSWPVRTPRPVASKLAADTPLLTGQRVLDALFPSVLGGTCAIPGAFGCGKTVISQALSKYSNSDAVVYVGCGERGNEMAEVLMDFPQLTMTLPDGREESVMKRTTLVANTSNMPVAAREASIYTGITIAEYFRDMGYNVSMMADSTSRWAEALREISGRLAEMPADSGYPAYLAARLASFYERAGKVKCLGGPERNGSVTIVGAVSPPGGDFSDPVTSATLSIVQVFWGLDKKLAQRKHFPSVNWLISYSKYSTALESFYEKFDPDFINIRTKAREVLQREDDLNEIVQLVGKDALAEGDKITLETAKLLREDYLAQNAFTPYDKFCPFYKSVWMMRNIIHFYNLANQAVERAAGMDGQKITYTLIKHRLGDLFYRLVSQKFEDPAEGEDTLVEKFKKLYDDLNAGFRALEDETR F63CAA1A9294CD4F vatc_arath V-type proton ATPase subunit C VHA-C DET3 F13K23.9 At1g12840 Vacuolar proton pump subunit C VATC Vacuolar H(+)-ATPase subunit C protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MTSRYWVVSLPVKDSASSLWNRLQEQISKHSFDTPVYRFNIPNLRVGTLDSLLALGDDLLKSNSFVEGVSQKIRRQIEELERISGVESNALTVDGVPVDSYLTRFVWDEAKYPTMSPLKEVVDNIQSQVAKIEDDLKVRVAEYNNIRGQLNAINRKQSGSLAVRDLSNLVKPEDIVESEHLVTLLAVVPKYSQKDWLACYETLTDYVVPRSSKKLFEDNEYALYTVTLFTRVADNFRIAAREKGFQVRDFEQSVEAQETRKQELAKLVQDQESLRSSLLQWCYTSYGEVFSSWMHFCAVRTFAESIMRYGLPPAFLACVLSPAVKSEKKVRSILERLCDSTNSLYWKSEEDAGAMAGLAGDSETHPYVSFTINLA A50F11D168F31C97 vate1_arath V-type proton ATPase subunit E1 TUF EMB2448 F2P3.10 T22B4.130 At4g11150 VHA-E1 Protein EMBRYO DEFECTIVE 2448 TUFF VATE Vacuolar proton pump subunit E1 Vacuolar H(+)-ATPase subunit E isoform 1 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MNDGDVSRQIQQMVRFIRQEAEEKANEISVSAEEEFNIEKLQLVEAEKKKIRQDYEKKEKQADVRKKIDYSMQLNASRIKVLQAQDDIVNAMKDQAAKDLLNVSRDEYAYKQLLKDLIVQCLLRLKEPSVLLRCREEDLGLVEAVLDDAKEEYAGKAKVHAPEVAVDTKIFLPPPPKSNDPHGLHCSGGVVLASRDGKIVCENTLDARLDVAFRMKLPVIRKSLFGQVTA 633E1D8D78A0889B sos2-at1g78900 1 5 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 3 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 4 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait c-terminal tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range association association Table 1 SOS2 interaction with and regulation of V-ATPase. NTAP-SOS2 purification identified five subunits (A, B1/2/3, C, E1, and G1) of the V-ATPase peripheral (cytoplasmic) sector as being components of a SOS2-containing protein complex. The authors noted "in the NTAP-SOS2 complexes purified from salt-stressed plants we observed a 4.1-fold increase in the relative amount of VHA-C, a 3.6-fold increase in VHA-A, and a 2.5-fold increase in VHA-B compared to the control". The authors cocluded "the consistently higher levels of several V-ATPase peripheral subunits associated with SOS2 in salt-stressed plants suggested that interaction of SOS2 with the V-ATPase complex is enhanced by salt stress". It is unclear whether the authors could identify the presence AtVHA-B1,B2 and B3 (At1g76030/At4g38510/At1g20260) from the LC-MS/MS data or if they could not tell which isoform was present. In this instance all three isoforms have been included into the entry with a caution for the user. Salt stress treatment as described in the text. sos2-at1g76030 2 7 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 6 unspecified role unspecified role bait bait Q9LDI3:p.Lys40Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K N Wed May 02 19:38:19 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys40asn' Tue Jun 30 18:09:41 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys40Asn' physical association physical association Figure 4B To confirm the interaction between VHA-B proteins and SOS2, the entire open reading frames of VHA-B1 and VHA-B2 were fused with the GAL4 activation domain in the prey plasmid pACT2 and cotransformed with the bait plasmid pAS-SOS2, pAS-SOS2 K40N, or pAS-SOS1 (33) into the yeast strain Y190. SOS2 interacted with both of the VHA-B subunits, as indicated by the β-galactosidase activity produced sos2-at4g38510 2 4 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 6 unspecified role unspecified role bait bait Q9LDI3:p.Lys40Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K N Wed May 02 20:28:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys40asn' Tue Jun 30 16:57:48 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys40Asn' physical association physical association Figure 4A To confirm the interaction between VHA-B proteins and SOS2, the entire open reading frames of VHA-B1 and VHA-B2 were fused with the GAL4 activation domain in the prey plasmid pACT2 and cotransformed with the bait plasmid pAS-SOS2, pAS-SOS2 K40N, or pAS-SOS1 (33) into the yeast strain Y190. SOS2 interacted with both of the VHA-B subunits, as indicated by the β-galactosidase activity produced