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/ RAP1 is essential for silencing telomeric variant surface glycoprotein genes in Trypanosoma brucei. RAP1 is essential for silencing telomeric variant surface glycoprotein genes in Trypanosoma brucei. Cell (0092-8674) 2009 imex curation Yang X., Figueiredo LM., Espinal A., Okubo E., Li B. b.li37@csuohio.edu 21-Dec-2010: Contacted by JYOTI. A GAD cDNA library generated with T. brucei 427 cDNA davet Only protein-protein interactions 6 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Yang X., Figueiredo LM., Espinal A., Okubo E., Li B. b.li37@csuohio.edu Accepted 2009-JUL-2 by DAVET. A GAD cDNA library generated with T. brucei 427 cDNA Cell (0092-8674) 2009 davet Only protein-protein interactions 6 imex curation RAP1 is essential for silencing telomeric variant surface glycoprotein genes in Trypanosoma brucei. RAP1 is essential for silencing telomeric variant surface glycoprotein genes in Trypanosoma brucei. Cell (0092-8674) 2009 imex curation Yang X., Figueiredo LM., Espinal A., Okubo E., Li B. b.li37@csuohio.edu 21-Dec-2010: Contacted by JYOTI. A GAD cDNA library generated with T. brucei 427 cDNA davet Only protein-protein interactions 6 9tryp Trypanosoma brucei fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Yang X., Figueiredo LM., Espinal A., Okubo E., Li B. b.li37@csuohio.edu Accepted 2009-JUL-2 by DAVET. 21-Dec-2010: Contacted by JYOTI. A GAD cDNA library generated with T. brucei 427 cDNA Cell (0092-8674) 2009 davet Only protein-protein interactions 6 imex curation RAP1 is essential for silencing telomeric variant surface glycoprotein genes in Trypanosoma brucei. RAP1 is essential for silencing telomeric variant surface glycoprotein genes in Trypanosoma brucei. Cell (0092-8674) 2009 imex curation Yang X., Figueiredo LM., Espinal A., Okubo E., Li B. b.li37@csuohio.edu 21-Dec-2010: Contacted by JYOTI. A GAD cDNA library generated with T. brucei 427 cDNA davet Only protein-protein interactions 6 9tryp Trypanosoma brucei anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Yang X., Figueiredo LM., Espinal A., Okubo E., Li B. b.li37@csuohio.edu Accepted 2009-JUL-2 by DAVET. 21-Dec-2010: Contacted by JYOTI. A GAD cDNA library generated with T. brucei 427 cDNA Cell (0092-8674) 2009 davet Only protein-protein interactions 6 imex curation RAP1 is essential for silencing telomeric variant surface glycoprotein genes in Trypanosoma brucei. RAP1 is essential for silencing telomeric variant surface glycoprotein genes in Trypanosoma brucei. Cell (0092-8674) 2009 imex curation Yang X., Figueiredo LM., Espinal A., Okubo E., Li B. b.li37@csuohio.edu 21-Dec-2010: Contacted by JYOTI. A GAD cDNA library generated with T. brucei 427 cDNA davet Only protein-protein interactions 6 9tryp Trypanosoma brucei anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Yang X., Figueiredo LM., Espinal A., Okubo E., Li B. b.li37@csuohio.edu Accepted 2009-JUL-2 by DAVET. 21-Dec-2010: Contacted by JYOTI. A GAD cDNA library generated with T. brucei 427 cDNA Cell (0092-8674) 2009 davet Only protein-protein interactions 6 imex curation RAP1 is essential for silencing telomeric variant surface glycoprotein genes in Trypanosoma brucei. RAP1 is essential for silencing telomeric variant surface glycoprotein genes in Trypanosoma brucei. Cell (0092-8674) 2009 imex curation Yang X., Figueiredo LM., Espinal A., Okubo E., Li B. b.li37@csuohio.edu 21-Dec-2010: Contacted by JYOTI. A GAD cDNA library generated with T. brucei 427 cDNA davet Only protein-protein interactions 6 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 Yang X., Figueiredo LM., Espinal A., Okubo E., Li B. b.li37@csuohio.edu Accepted 2009-JUL-2 by DAVET. 21-Dec-2010: Contacted by JYOTI. A GAD cDNA library generated with T. brucei 427 cDNA Cell (0092-8674) 2009 davet Only protein-protein interactions 6 imex curation q4vhg6_9tryp TRF protein protein 9tryp Trypanosoma brucei MYCHAGVPSTSPSPTDFSEVHKILCGHIKQGNNFSVAAAVALEPLHGSSEFWRTCGYRSGIVSALLASSLGVQRETDQLSHLEPMTVADVYAMFRHVVDENAHLRQIIARLESGECDPDDGSGGITSSAERLRSLISEGYDALTGRSSPCMGEDDCGQTPGKWKTCSSGAAGTKRRRDSPEVTERAYVRPESPVPASETLSPAANSGDDTVDEAEQNNALDVLEQLPALPGTTRTVSSFDAVGEMVNRGAGGPSLHPTNPSEFRVGSERGSTIETHSSSELSNFANVGVGTSSGKQKRYKFSASEDEAIIKGLARFTKGQQRFQQIYYAYRSVWHPARTVSQLYDHWRGTLRYKVIQQQGYRGKNSVAARSPEKASNMENNE A1C4A630D42C4FCD q387l4_tryb2 Tb11.03.0760 protein protein tryb2 Trypanosoma brucei brucei (strain 927/4 GUTat10.1) MTDVDGGSGASWKRLRFFVCADTLLTFQPVDSSHLEGRPGQPEQRYLLVDGEYRPITSYPTSSLFLETLKLAGSTVVVIAPWKRAKTKKLLSLFGWTAFDEIRSIHPEPGRDKLHLACIGCSADEYSTSVIVATSRADWSPSMYPQLVEVKHGVNQVPHSLALLRALSSCLRLAEWQVRLPHVPISSCATLCRSRLFSHCRFFLEPVVSEDATVALSINEHGGYIVTRREDATHCVVSGSLPNLPGDISNLPPGGCAAKAVKECSVLPTSAEGRSAEPKSAKVDTDSVSLVKNEVDESDDSIKDEFSYSAGNAATAMVECGDEGVWNEENADDEDPVTMIPPGTFSEDDEDDAPGGLEEADGETVTTVTTQWLKDCAEGLFMYPPQVSHTSKDGSDWNITGLLLATILPTSDAEPLLLSSFNTFAERSGFEPLQLNSADEGGLAEVMQRQWGGVLCMEEKDGDFLLSQTNIYTEAFSDEMDKHLTFVANRFCEAWKDAVQRRRALLGSDAGTQTTVSITVTFGTNTEVEVAPSVQPPPPPPPAPQPTTASAAANEGSKESATPALPFLMRALGPAEWAKEVEAPRSPPPSAVPDIPLKQDPVPSGEGVATAENSVGNDQKESTSRQVAGDAGDTANVAEATEEIAALDQPFEKCFIPTEALGSDREGLDRTQLERQLPFRNYPIKLNVSKSGIFCQFPTVSDAKRFYEEGTVEILNRSLPIKPVFEKRNETVAPAERKRRRSVSPGGVHPQTAAVSALSRRGIQGATYARTQRSRTDNGWKAEGDRNPFVLTEEENSLLTSLGITYTYALENVPLLEHYAQKDLSSKVYRKEKGVLVKMLEKLKAVKGTTSKAPQ E7E5A8BD98C36CD9 trb-rap1-1 1 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association we carried out a yeast two-hybrid screen using tbTRF as bait. More than ten tbTRF-interacting candidates were identified, including the conserved hypothetical protein Tb11.03.0760. trb-rap1-2 5 6 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-2338864]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 399-399, sequence length 382 [EBI-2338864]2-399 7 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 13 15 physical association physical association TbRAP1 interacts with tbTRF in yeast two-hybrid analysis. Fig. 2A trb-trb-1 5 6 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-2338880]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 399-399, sequence length 382 [EBI-2338880]2-399 6 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-2338871]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 399-399, sequence length 382 [EBI-2338871]2-399 20 18 physical association physical association Compared with tbTRF self-interaction Fig. 2A trb-rap1-3 3 7 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey physical association physical association The endogenous F2H-tbRAP1 partially coIP with the endogenous tbTRF. Fig. 2B trb-rap1-4 4 6 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association The endogenous F2H-tbRAP1 partially coIP with the endogenous tbTRF. Fig. 2B trb-rap1-5 2 6 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component ty1 region tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization Ty1-tbRAP1 partially colocalizes with tbTRF. Fig. 2C