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/ Molecular chaperone Hsp90 stabilizes Pih1/Nop17 to maintain R2TP complex activity that regulates snoRNA accumulation. Molecular chaperone Hsp90 stabilizes Pih1/Nop17 to maintain R2TP complex activity that regulates snoRNA accumulation. J. Cell Biol. (0021-9525) 2008 imex curation Zhao R., Kakihara Y., Gribun A., Huen J., Yang G., Khanna M., Costanzo M., Brost RL., Boone C., Hughes TR., Yip CM., Houry WA. walid.houry@utoronto.ca 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Zhao R., Kakihara Y., Gribun A., Huen J., Yang G., Khanna M., Costanzo M., Brost RL., Boone C., Hughes TR., Yip CM., Houry WA. Superdex 200 HR 10/30 column walid.houry@utoronto.ca Accepted 2010-MAR-12 by JYOTI. 23-Feb-2010: Contacted by JYOTI. J. Cell Biol. (0021-9525) 2008 jyoti Only protein-protein interactions imex curation Molecular chaperone Hsp90 stabilizes Pih1/Nop17 to maintain R2TP complex activity that regulates snoRNA accumulation. Molecular chaperone Hsp90 stabilizes Pih1/Nop17 to maintain R2TP complex activity that regulates snoRNA accumulation. J. Cell Biol. (0021-9525) 2008 imex curation Zhao R., Kakihara Y., Gribun A., Huen J., Yang G., Khanna M., Costanzo M., Brost RL., Boone C., Hughes TR., Yip CM., Houry WA. walid.houry@utoronto.ca 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Zhao R., Kakihara Y., Gribun A., Huen J., Yang G., Khanna M., Costanzo M., Brost RL., Boone C., Hughes TR., Yip CM., Houry WA. walid.houry@utoronto.ca Accepted 2010-MAR-12 by JYOTI. 23-Feb-2010: Contacted by JYOTI. J. Cell Biol. (0021-9525) 2008 jyoti Only protein-protein interactions imex curation Molecular chaperone Hsp90 stabilizes Pih1/Nop17 to maintain R2TP complex activity that regulates snoRNA accumulation. Molecular chaperone Hsp90 stabilizes Pih1/Nop17 to maintain R2TP complex activity that regulates snoRNA accumulation. J. Cell Biol. (0021-9525) 2008 imex curation Zhao R., Kakihara Y., Gribun A., Huen J., Yang G., Khanna M., Costanzo M., Brost RL., Boone C., Hughes TR., Yip CM., Houry WA. walid.houry@utoronto.ca 23-Feb-2010: Contacted by JYOTI. jyoti 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 Zhao R., Kakihara Y., Gribun A., Huen J., Yang G., Khanna M., Costanzo M., Brost RL., Boone C., Hughes TR., Yip CM., Houry WA. walid.houry@utoronto.ca Accepted 2010-MAR-12 by JYOTI. 23-Feb-2010: Contacted by JYOTI. J. Cell Biol. (0021-9525) 2008 jyoti Only protein-protein interactions imex curation Molecular chaperone Hsp90 stabilizes Pih1/Nop17 to maintain R2TP complex activity that regulates snoRNA accumulation. Molecular chaperone Hsp90 stabilizes Pih1/Nop17 to maintain R2TP complex activity that regulates snoRNA accumulation. J. Cell Biol. (0021-9525) 2008 imex curation Zhao R., Kakihara Y., Gribun A., Huen J., Yang G., Khanna M., Costanzo M., Brost RL., Boone C., Hughes TR., Yip CM., Houry WA. walid.houry@utoronto.ca 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip ms participant Identification by mass spectrometry ms participant Zhao R., Kakihara Y., Gribun A., Huen J., Yang G., Khanna M., Costanzo M., Brost RL., Boone C., Hughes TR., Yip CM., Houry WA. walid.houry@utoronto.ca Accepted 2010-MAR-12 by JYOTI. 23-Feb-2010: Contacted by JYOTI. J. Cell Biol. (0021-9525) 2008 jyoti Only protein-protein interactions imex curation hsp82_yeast ATP-dependent molecular chaperone HSP82 HSP90 YPL240C HSP82 Heat shock protein Hsp90 heat-inducible isoform 82 kDa heat shock protein protein protein yeast Saccharomyces cerevisiae Baker's yeast MASETFEFQAEITQLMSLIINTVYSNKEIFLRELISNASDALDKIRYKSLSDPKQLETEPDLFIRITPKPEQKVLEIRDSGIGMTKAELINNLGTIAKSGTKAFMEALSAGADVSMIGQFGVGFYSLFLVADRVQVISKSNDDEQYIWESNAGGSFTVTLDEVNERIGRGTILRLFLKDDQLEYLEEKRIKEVIKRHSEFVAYPIQLVVTKEVEKEVPIPEEEKKDEEKKDEEKKDEDDKKPKLEEVDEEEEKKPKTKKVKEEVQEIEELNKTKPLWTRNPSDITQEEYNAFYKSISNDWEDPLYVKHFSVEGQLEFRAILFIPKRAPFDLFESKKKKNNIKLYVRRVFITDEAEDLIPEWLSFVKGVVDSEDLPLNLSREMLQQNKIMKVIRKNIVKKLIEAFNEIAEDSEQFEKFYSAFSKNIKLGVHEDTQNRAALAKLLRYNSTKSVDELTSLTDYVTRMPEHQKNIYYITGESLKAVEKSPFLDALKAKNFEVLFLTDPIDEYAFTQLKEFEGKTLVDITKDFELEETDEEKAEREKEIKEYEPLTKALKEILGDQVEKVVVSYKLLDAPAAIRTGQFGWSANMERIMKAQALRDSSMSSYMSSKKTFEISPKSPIIKELKKRVDEGGAQDKTVKDLTKLLYETALLTSGFSLDEPTSFASRINRLISLGLNIDEDEETETAPEASTAAPVEEVPADTEMEEVD D7C35676D668FB63 D7C35676D668FB63 sti1_yeast Heat shock protein STI1 STI1 OR26.17 YOR027W protein protein yeast Saccharomyces cerevisiae Baker's yeast MSLTADEYKQQGNAAFTAKDYDKAIELFTKAIEVSETPNHVLYSNRSACYTSLKKFSDALNDANECVKINPSWSKGYNRLGAAHLGLGDLDEAESNYKKALELDASNKAAKEGLDQVHRTQQARQAQPDLGLTQLFADPNLIENLKKNPKTSEMMKDPQLVAKLIGYKQNPQAIGQDLFTDPRLMTIMATLMGVDLNMDDINQSNSMPKEPETSKSTEQKKDAEPQSDSTTSKENSSKAPQKEESKESEPMEVDEDDSKIEADKEKAEGNKFYKARQFDEAIEHYNKAWELHKDITYLNNRAAAEYEKGEYETAISTLNDAVEQGREMRADYKVISKSFARIGNAYHKLGDLKKTIEYYQKSLTEHRTADILTKLRNAEKELKKAEAEAYVNPEKAEEARLEGKEYFTKSDWPNAVKAYTEMIKRAPEDARGYSNRAAALAKLMSFPEAIADCNKAIEKDPNFVRAYIRKATAQIAVKEYASALETLDAARTKDAEVNNGSSAREIDQLYYKASQQRFQPGTSNETPEETYQRAMKDPEVAAIMQDPVMQSILQQAQQNPAALQEHMKNPEVFKKIQTLIAAGIIRTGR CF8A85C2BBD3C44C CF8A85C2BBD3C44C ruvb2_yeast RuvB-like protein 2 TIP49-homology protein 2 TIP49b homolog RVB2 TIH2 TIP49B YPL235W P1060 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSIQTSDPNETSDLKSLSLIAAHSHITGLGLDENLQPRPTSEGMVGQLQARRAAGVILKMVQNGTIAGRAVLVAGPPSTGKTALAMGVSQSLGKDVPFTAIAGSEIFSLELSKTEALTQAFRKSIGIKIKEETELIEGEVVEIQIDRSITGGHKQGKLTIKTTDMETIYELGNKMIDGLTKEKVLAGDVISIDKASGKITKLGRSFARSRDYDAMGADTRFVQCPEGELQKRKTVVHTVSLHEIDVINSRTQGFLALFTGDTGEIRSEVRDQINTKVAEWKEEGKAEIVPGVLFIDEVHMLDIECFSFINRALEDEFAPIVMMATNRGVSKTRGTNYKSPHGLPLDLLDRSIIITTKSYNEQEIKTILSIRAQEEEVELSSDALDLLTKTGVETSLRYSSNLISVAQQIAMKRKNNTVEVEDVKRAYLLFLDSARSVKYVQENESQYIDDQGNVQISIAKSADPDAMDTTE 0D8EFBA7EC711AB8 0D8EFBA7EC711AB8 ruvb1_yeast RuvB-like protein 1 RVB1 TIP49-homology protein 1 TIP49a homolog TIH1 TIP49A YDR190C protein protein yeast Saccharomyces cerevisiae Baker's yeast MVAISEVKENPGVNSSNSGAVTRTAAHTHIKGLGLDESGVAKRVEGGFVGQIEAREACGVIVDLIKAKKMSGRAILLAGGPSTGKTALALAISQELGPKVPFCPLVGSELYSVEVKKTETLMENFRRAIGLRIKETKEVYEGEVTELTPEDAENPLGGYGKTISHVIVGLKSAKGTKTLRLDPTIYESIQREKVSIGDVIYIEANTGAVKRVGRSDAYATEFDLETEEYVPLPKGEVHKKKEIVQDVTLHDLDVANARPQGGQDVISMMGQLLKPKKTEITEKLRQEVNKVVAKYIDQGVAELIPGVLFIDEVNMLDIEIFTYLNKALESNIAPVVVLASNRGMTTVRGTEDVISPHGVPPDLIDRLLIVRTLPYDKDEIRTIIERRATVERLQVESSALDLLATMGTETSLRYALQLLAPCGILAQTSNRKEIVVNDVNEAKLLFLDAKRSTKILETSANYL 93D6ECB7006AF0F2 93D6ECB7006AF0F2 pih1_yeast Protein interacting with Hsp90 1 PIH1 NOP17 Nucleolar protein 17 YHR034C protein protein yeast Saccharomyces cerevisiae Baker's yeast MADFLLRPIKQRHRNEDKYVSVDAADGSVSKIEPIADFVIKTKLLSANGPEKLQDGRKVFINVCHSPLVPKPEVDFNARIVFPLIIQNEWEIPIITSCYRMDHDKKGQECYVWDCCINSDCSRWICDDIQLREILVEWCLESCEIRDSVVLCRDRIAFPKMKKKGAELPALEVLNDELHQDYKAKMHKIIEEEAGDPMSILRGRNDDGDDNNDPDDGTLPPLFPIENKISGAKIEEIDKNEIAHRNLKQAPAPAPAPHEQQEDVPEYEVKMKRFKGAAYKLRILIENKAPNSKPDRFSPSYNFAENILYINGKLSIPLPRDIVVNAADIKIFHIRKERTLYIYI 02C06D0A113B1B3B 02C06D0A113B1B3B tah1_yeast TPR repeat-containing protein associated with Hsp90 YCR60W YCR060W TAH1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSQFEKQKEQGNSLFKQGLYREAVHCYDQLITAQPQNPVGYSNKAMALIKLGEYTQAIQMCQQGLRYTSTAEHVAIRSKLQYRLELAQGAVGSVQIPVVEVDELPEGYDRS 3738DE6F227F803A 3738DE6F227F803A tah1-pih1-1 2 9 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role bait bait n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Surprisingly, Tah1-C is sufficient to bind Pih1 Fig. 1B tah1-hsp82-1 2 5 unspecified role unspecified role prey prey mc region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role bait bait n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) direct interaction direct interaction Furthermore, full-length Tah1 interacted with Hsp82-MC but not with the Hsp82-NM fragment Fig. 1B sti1-hsp82-2 2 6 unspecified role unspecified role bait bait n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined tpr region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role prey prey mc region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 24 26 direct interaction direct interaction As a positive control, the TPR2a domain from Sti1 (Fig. 1 A) interacted with Hsp82-MC but not with Hsp82-NM Fig. 1B pih1-hsp82-1 3 9 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey mc region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association The interaction between Hsp82-C and Pih1 was characterized using yeast 2H. Fig. 1C Interaction occurs in TAH1-/- cells. hsp82-1 1 5 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association 1E Hsp82 eluted at the position of a dimer tah1-pih1-2 1 9 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association however, a complex of Tah1 and Pih1 was evident Fig. 1E rvb1-rvb2-1 1 8 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association When Rvb1 and Rvb2 were incubated together for 4 h before being separated on the SE column, the two helicases formed a complex of an apparent molecular mass of ~300-600 kD in the presence or absence of ADP Fig. 2A rvb1-rvb2-2 1 9 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association When both Tah1 and Pih1 were mixed with the Rvb1/2 complex, coelution of Rvb1, Rvb2, Tah1, and Pih1 was observed over several fractions, indicating that the complex might be dynamic Fig. 2B rvb1-rvb2-3 1 9 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association When Tah1 or Pih1 was added individually to the Rvb1/2 complex, Pih1 comigrated with Rvb1/2 Fig. 2B rvb1-rvb2-4 4 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Immunoprecipitations with anti-FLAG antibody indicated that Rvb1 and Rvb2 are associated with Tah1 and Pih1 when isolated from WT cells (Fig. 2 C, left) Fig. 2C Interaction is not observed in pih1-/- cells. rvb1-rvb2-5 4 10 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey association association Immunoprecipitations with anti-FLAG antibody indicated that Rvb1 and Rvb2 are associated with Tah1 and Pih1 when isolated from WT cells (Fig. 2 C, left) Fig. 2C, 2D rvb1-rvb2-6 4 9 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey association association 2D Interaction observed in Tah1-/- cells