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/ RORalpha-mediated Purkinje cell development determines disease severity in adult SCA1 mice. RORalpha-mediated Purkinje cell development determines disease severity in adult SCA1 mice. Cell (0092-8674) 2006 imex curation Serra HG., Duvick L., Zu T., Carlson K., Stevens S., Jorgensen N., Lysholm A., Burright E., Zoghbi HY., Clark HB., Andresen JM., Orr HT. A parasagittal cerebellar section from a 12 week old SCA1[30Q] heterozygous mouse was simultaneously stained with antibodies against ATXN1 and RORalpha. orrxx002@umn.edu 08-DEC-2006: Contacted by JYOTI. Only protein-protein interactions 10 mouse-brain-cerebellum Mus musculus brain cerebellum brain_cerebellum [Cerebellar] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Serra HG., Duvick L., Zu T., Carlson K., Stevens S., Jorgensen N., Lysholm A., Burright E., Zoghbi HY., Clark HB., Andresen JM., Orr HT. A parasagittal cerebellar section from a 12 week old SCA1[30Q] heterozygous mouse was simultaneously stained with antibodies against ATXN1 and RORalpha. orrxx002@umn.edu Accepted 2006-NOV-27 by JYOTI. 08-DEC-2006: Contacted by JYOTI. Cell (0092-8674) 2006 Only protein-protein interactions 10 imex curation RORalpha-mediated Purkinje cell development determines disease severity in adult SCA1 mice. RORalpha-mediated Purkinje cell development determines disease severity in adult SCA1 mice. Cell (0092-8674) 2006 imex curation Serra HG., Duvick L., Zu T., Carlson K., Stevens S., Jorgensen N., Lysholm A., Burright E., Zoghbi HY., Clark HB., Andresen JM., Orr HT. A parasagittal cerebellar section from a 12 week old SCA1[30Q] heterozygous mouse was simultaneously stained with antibodies against ATXN1 and RORalpha. orrxx002@umn.edu 08-DEC-2006: Contacted by JYOTI. Only protein-protein interactions 10 in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Serra HG., Duvick L., Zu T., Carlson K., Stevens S., Jorgensen N., Lysholm A., Burright E., Zoghbi HY., Clark HB., Andresen JM., Orr HT. This experiment relates to a negative interaction. A parasagittal cerebellar section from a 12 week old SCA1[30Q] heterozygous mouse was simultaneously stained with antibodies against ATXN1 and RORalpha. orrxx002@umn.edu Accepted 2006-NOV-27 by JYOTI. 08-DEC-2006: Contacted by JYOTI. Cell (0092-8674) 2006 Only protein-protein interactions 10 imex curation RORalpha-mediated Purkinje cell development determines disease severity in adult SCA1 mice. RORalpha-mediated Purkinje cell development determines disease severity in adult SCA1 mice. Cell (0092-8674) 2006 imex curation Serra HG., Duvick L., Zu T., Carlson K., Stevens S., Jorgensen N., Lysholm A., Burright E., Zoghbi HY., Clark HB., Andresen JM., Orr HT. A parasagittal cerebellar section from a 12 week old SCA1[30Q] heterozygous mouse was simultaneously stained with antibodies against ATXN1 and RORalpha. orrxx002@umn.edu 08-DEC-2006: Contacted by JYOTI. Only protein-protein interactions 10 chlae-cos_1 Chlorocebus aethiops COS-1 cells Grivet cos_1 CV-1 Simian cells (cercopithecus aethiops) transformed by SV40 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Serra HG., Duvick L., Zu T., Carlson K., Stevens S., Jorgensen N., Lysholm A., Burright E., Zoghbi HY., Clark HB., Andresen JM., Orr HT. A parasagittal cerebellar section from a 12 week old SCA1[30Q] heterozygous mouse was simultaneously stained with antibodies against ATXN1 and RORalpha. orrxx002@umn.edu Accepted 2006-NOV-27 by JYOTI. 08-DEC-2006: Contacted by JYOTI. Cell (0092-8674) 2006 Only protein-protein interactions 10 imex curation RORalpha-mediated Purkinje cell development determines disease severity in adult SCA1 mice. RORalpha-mediated Purkinje cell development determines disease severity in adult SCA1 mice. Cell (0092-8674) 2006 imex curation Serra HG., Duvick L., Zu T., Carlson K., Stevens S., Jorgensen N., Lysholm A., Burright E., Zoghbi HY., Clark HB., Andresen JM., Orr HT. A parasagittal cerebellar section from a 12 week old SCA1[30Q] heterozygous mouse was simultaneously stained with antibodies against ATXN1 and RORalpha. orrxx002@umn.edu 08-DEC-2006: Contacted by JYOTI. Only protein-protein interactions 10 mouse-purkinje cell Mus musculus brain-purkinje_cell Purkinje cell Purkinje cells can be found either as 1] Purkinje fibers in the inner ventricular walls of the heart, that conduct an electrical stimulus that enables the heart to contract in a coordinated fashion - or 2]Purkinje neurons, a class of GABAergic neurons located in the cerebellar cortex. confocal microscopy confocal microscopy immunostaining immunostaining Serra HG., Duvick L., Zu T., Carlson K., Stevens S., Jorgensen N., Lysholm A., Burright E., Zoghbi HY., Clark HB., Andresen JM., Orr HT. A parasagittal cerebellar section from a 12 week old SCA1[30Q] heterozygous mouse was simultaneously stained with antibodies against ATXN1 and RORalpha. orrxx002@umn.edu Accepted 2006-NOV-27 by JYOTI. 08-DEC-2006: Contacted by JYOTI. Cell (0092-8674) 2006 Only protein-protein interactions 10 imex curation kat5_human Histone acetyltransferase KAT5 KAT5 TIP60 HTATIP Lysine acetyltransferase 5 cPLA(2)-interacting protein Histone acetyltransferase HTATIP 60 kDa Tat-interactive protein protein protein human Homo sapiens Human MAEVGEIIEGCRLPVLRRNQDNEDEWPLAEILSVKDISGRKLFYVHYIDFNKRLDEWVTHERLDLKKIQFPKKEAKTPTKNGLPGSRPGSPEREVPASAQASGKTLPIPVQITLRFNLPKEREAIPGGEPDQPLSSSSCLQPNHRSTKRKVEVVSPATPVPSETAPASVFPQNGAARRAVAAQPGRKRKSNCLGTDEDSQDSSDGIPSAPRMTGSLVSDRSHDDIVTRMKNIECIELGRHRLKPWYFSPYPQELTTLPVLYLCEFCLKYGRSLKCLQRHLTKCDLRHPPGNEIYRKGTISFFEIDGRKNKSYSQNLCLLAKCFLDHKTLYYDTDPFLFYVMTEYDCKGFHIVGYFSKEKESTEDYNVACILTLPPYQRRGYGKLLIEFSYELSKVEGKTGTPEKPLSDLGLLSYRSYWSQTILEILMGLKSESGERPQITINEISEITSIKKEDVISTLQYLNLINYYKGQYILTLSEDIVDGHERAMLKRLLRIDSKCLHFTPKDWSKRGKW DIP protein Q92993 original sequence version: 153 DIP protein Q92993 original sequence version: 155 DIP protein Q92993 original sequence version: 86 DIP protein Q92993 original sequence version: 149 DIP protein Q92993-1 original sequence version: 129 DIP protein Q92993 original sequence version: 144 DIP protein Q92993 original sequence version: 173 63724F5E10B957D5 atx1_mouse Ataxin-1 Spinocerebellar ataxia type 1 protein homolog Atxn1 Sca1 protein protein mouse Mus musculus Mouse MKSNQERSNECLPPKKREIPATSRPSEEKATALPSDNHCVEGVAWLPSTPGIRGHGGGRHGSAGTSGEHGLQGMGLHKALSAGLDYSPPSAPRSVPTANTLPTVYPPPQSGTPVSPVQYAHLSHTFQFIGSSQYSGPYAGFIPSQLISPSGNPVTSAVASAAGATTPSQRSQLEAYSTLLANMGSLSQAPGHKVEPPPQQHLSRAAGLVNPGSPPPPTQQNQYIHISSSPQSSGRATSPPPIPVHLHPHQTMIPHTLTLGPSSQVVVQYSDAGGHFVPRESTKKAESSRLQQAMQAKEVLNGEMEKSRRYGASSSVELSLGKASSKSVPHPYESRHVVVHPSPADYSSRDTSGVRGSVMVLPNSSTPSADLEAQQTTHREASPSTLNDKSGLHLGKPGHRSYALSPHTVIQTTHSASEPLPVGLPATAFYAGTQPPVIGYLSGQQQAITYAGGLPQHLVIPGNQPLLIPVGSPDMDTPGAASAIVTSSPQFAAVPHTFVTTALPKSENFNPEALVTQAAYPAMVQAQIHLPVVQSVASPTTASPTLPPYFMKGSIIQLANGELKKVEDLKTEDFIQSAEISNDLKIDSSTVERIEESHSPGVAVIQFAVGEHRAQVSVEVLVEYPFFVFGQGWSSCCPERTSQLFDLPCSKLSVGDVCISLTLKNLKNGSVKKGQPVDPASVLLKQAKTDSLAGSRHRYAEQENGINQGSAQVLSENGELKFPEKIGLPAAPFLSKIEPSKPTATRKRRWSAPETRKLEKSEDEPPLTLPKPSLIPQEVKICIEGRSNVGK DIP protein P54254 original sequence version: 112 1F87A5D65527D550 atx1_human Ataxin-1 ATXN1 ATX1 SCA1 Spinocerebellar ataxia type 1 protein protein protein human Homo sapiens Human MKSNQERSNECLPPKKREIPATSRSSEEKAPTLPSDNHRVEGTAWLPGNPGGRGHGGGRHGPAGTSVELGLQQGIGLHKALSTGLDYSPPSAPRSVPVATTLPAAYATPQPGTPVSPVQYAHLPHTFQFIGSSQYSGTYASFIPSQLIPPTANPVTSAVASAAGATTPSQRSQLEAYSTLLANMGSLSQTPGHKAEQQQQQQQQQQQQHQHQQQQQQQQQQQQQQHLSRAPGLITPGSPPPAQQNQYVHISSSPQNTGRTASPPAIPVHLHPHQTMIPHTLTLGPPSQVVMQYADSGSHFVPREATKKAESSRLQQAIQAKEVLNGEMEKSRRYGAPSSADLGLGKAGGKSVPHPYESRHVVVHPSPSDYSSRDPSGVRASVMVLPNSNTPAADLEVQQATHREASPSTLNDKSGLHLGKPGHRSYALSPHTVIQTTHSASEPLPVGLPATAFYAGTQPPVIGYLSGQQQAITYAGSLPQHLVIPGTQPLLIPVGSTDMEASGAAPAIVTSSPQFAAVPHTFVTTALPKSENFNPEALVTQAAYPAMVQAQIHLPVVQSVASPAAAPPTLPPYFMKGSIIQLANGELKKVEDLKTEDFIQSAEISNDLKIDSSTVERIEDSHSPGVAVIQFAVGEHRAQVSVEVLVEYPFFVFGQGWSSCCPERTSQLFDLPCSKLSVGDVCISLTLKNLKNGSVKKGQPVDPASVLLKHSKADGLAGSRHRYAEQENGINQGSAQMLSENGELKFPEKMGLPAAPFLTKIEPSKPAATRKRRWSAPESRKLEKSEDEPPLTLPKPSLIPQEVKICIEGRSNVGK DIP protein P54253 original sequence version: 102 DIP protein P54253 original sequence version: 135 657876F8FD19ECB2 rora_mouse Nuclear receptor ROR-alpha Retinoid-related orphan receptor-alpha Nuclear receptor RZR-alpha Nuclear receptor subfamily 1 group F member 1 Rora Nr1f1 Rzra RAR-related orphan receptor A protein protein mouse Mus musculus Mouse MESAPAAPDPAASEPGSSGSEAAAGSRETPLTQDTGRKSEAPGAGRRQSYASSSRGISVTKKTHTSQIEIIPCKICGDKSSGIHYGVITCEGCKGFFRRSQQSNATYSCPRQKNCLIDRTSRNRCQHCRLQKCLAVGMSRDAVKFGRMSKKQRDSLYAEVQKHRMQQQQRDHQQQPGEAEPLTPTYNISANGLTELHDDLSTYMDGHTPEGSKADSAVSSFYLDIQPSPDQSGLDINGIKPEPICDYTPASGFFPYCSFTNGETSPTVSMAELEHLAQNISKSHLETCQYLREELQQITWQTFLQEEIENYQNKQREVMWQLCAIKITEAIQYVVEFAKRIDGFMELCQNDQIVLLKAGSLEVVFIRMCRAFDSQNNTVYFDGKYASPDVFKSLGCEDFISFVFEFGKSLCSMHLTEDEIALFSAFVLMSADRSWLQEKVKIEKLQQKIQLALQHVLQKNHREDGILTKLICKVSTLRALCGRHTEKLMAFKAIYPDIVRLHFPPLYKELFTSEFEPAMQIDG DIP protein P51448 original sequence version: 116 A194E02E4D9D177E kat5_mouse Histone acetyltransferase KAT5 Kat5 Htatip Tip60 Lysine acetyltransferase 5 Histone acetyltransferase HTATIP 60 kDa Tat-interactive protein protein protein mouse Mus musculus Mouse MAEVGEIIEGCRLPVLRRNQDNEDEWPLAEILSVKDISGRKLFYVHYIDFNKRLDEWVTHERLDLKKIQFPKKEAKTPTKNGLPGSRPGSPEREVPASAQASGKTLPIPVQITLRFNLPKEREAIPGGEPDQPLSSSSCLQPNHRSTKRKVEVVSPATPVPSETAPASVFPQNGSARRAVAAQPGRKRKSNCLGTDEDSQDSSDGIPSAPRMTGSLVSDRSHDDIVTRMKNIECIELGRHRLKPWYFSPYPQELTTLPVLYLCEFCLKYGRSLKCLQRHLTKCDLRHPPGNEIYRKGTISFFEIDGRKNKSYSQNLCLLAKCFLDHKTLYYDTDPFLFYVMTEYDCKGFHIVGYFSKEKESTEDYNVACILTLPPYQRRGYGKLLIEFSYELSKVEGKTGTPEKPLSDLGLLSYRSYWSQTILEILMGLKSESGERPQITINEISEITSIKKEDVISTLQYLNLINYYKGQYILTLSEDIVDGHERAMLKRLLRIDSKCLHFTPKDWSKRGKW EACEE4D544C0DB60 atxn1-rora 4 6 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization ATXN1[30Q] and RORa proteins colocalized within the nuclei of Purkinje cells by confocal microscopy S2 rora-atxn1 1 6 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association An immunoprecipitation using an antibody against RORa coimmunoprecipitated ATXN1 from a lysate prepared from a normal mouse cerebellum 5 A This interaction did not take place in the Sca1-/- mice. atxn1-rora-2 1 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association RORa coimmunoprecipitated with the anti-ATXN1 antibody 11750 5 A This interaction did not take place in the Sca1-/- mice. htatip-atxn1-1 1 9 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association 5 C Coimmunoprecipitations using cerebella extracts from wild-type mice indicated that endogenous ATXN1 and Tip60 do interact atxn1-htatip 1 6 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association 5 C Coimmunoprecipitations using cerebella extracts from wild-type mice indicated that endogenous ATXN1 and Tip60 do interact atxn1-htatip-2 2 7 unspecified role unspecified role bait bait ecobb Escherichia coli (strain B / BL21) 9 unspecified role unspecified role prey prey mouse-brain-cerebellum Mus musculus brain cerebellum brain_cerebellum [Cerebellar] physical association physical association 5 D The ability of GSTATXN1 to pull down Tip60 from wild-type mouse cerebella extracts provided additional evidence that ATXN1 and Tip60 interact (Figure 5D). Furthermore, the GST-ATXN1 pull down results suggested that as the length of the polyglutamine tract increased from 30Q to 53Q to 82Q the amount of Tip60 pulled down increased. The amount of Tip60 pulled down increased as the length of the polyglutamine tract increased from 30 to 53 to 82. htatip-atxn1 3 5 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey physical association physical association 5 E Figure 5E shows that more ATXN1[82Q] was coimmunoprecipitated with Tip60 than was ATXN1[30Q] More ATXN1-82Q was coimmunoprecipitated with Tip60 than was ATXN1-30Q, further suggesting an effect of the polyglutamine sequence on the interaction of ATXN1 with Tip60. atxn1-htatip-3 2 5 unspecified role unspecified role prey prey in vitro In vitro 7 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 ecobb Escherichia coli (strain B / BL21) physical association physical association 5 F GST-ATXN1 was able to pull down in vitro transcribed/translated Tip60. However, this analysis failed to provide evidence of a polyglutamine tract effect. The increasing of the polyglutamine tract length in ATXN1 did not affect the amount of Tip60 pulled down by ATXN1. atxn1-htatip-1 3 7 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 c-terminal 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 physical association physical association 6 Of the ATXN1 fragments the C-terminal fragment (568-816) and the N-terminal fragment (1-360) interacted to a lesser extent with Tip60 than the two fragments containing aa 425-547 (fragments 250-547 and 425-689). The N-terminal fragment (1-360) of ATXN1 with the wild-type 30Q polyglutamine stretch did not interact strongly with Tip60 but the same fragment with the 82Q mutant polyglutamine stretch did. The two fragments of ATXN1 containing amino acids 425-547, fragment III and IV, interacted with Tip60 at least as well as intact ATXN1[30Q] did. In contrast, fragments lacking this portion of ATXN1 interacted to a lesser extent with Tip60 than did full-length ATXN1[30Q]. Notably, these include the N-terminal 360 amino-acid fragment containing the polyglutamine stretch and the C-terminal fragment spanning amino acids 568-816 containing the AXH domain AXH domain