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/ brasher-2000-1 The structure of mouse HP1 suggests a unique mode of single peptide recognition by the shadow chromo domain dimer. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Brasher SV., Smith BO., Fogh RH., Nietlispach D., Thiru A., Nielsen PR., Broadhurst RW., Ball LJ., Murzina NV., Laue ED. Purified proteins used. Mobility of the dimers on Superdex S75 gel-filteration column used to determine disruption of complex formation. 4, 5 nm@mole.bio.cam.ac.uk, e.d.laue@bioc.cam.ac.uk EMBO J. (0261-4189) 2000 brasher-2000-2 The structure of mouse HP1 suggests a unique mode of single peptide recognition by the shadow chromo domain dimer. in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Brasher SV., Smith BO., Fogh RH., Nietlispach D., Thiru A., Nielsen PR., Broadhurst RW., Ball LJ., Murzina NV., Laue ED. Purified proteins used. Mobility of the dimers on Superdex S75 gel-filteration column used to determine disruption of complex formation. 4, 5 nm@mole.bio.cam.ac.uk, e.d.laue@bioc.cam.ac.uk EMBO J. (0261-4189) 2000 brasher-2000-3 The structure of mouse HP1 suggests a unique mode of single peptide recognition by the shadow chromo domain dimer. in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Brasher SV., Smith BO., Fogh RH., Nietlispach D., Thiru A., Nielsen PR., Broadhurst RW., Ball LJ., Murzina NV., Laue ED. Purified proteins used. Mobility of the dimers on Superdex S75 gel-filteration column used to determine disruption of complex formation. 4, 5 nm@mole.bio.cam.ac.uk, e.d.laue@bioc.cam.ac.uk EMBO J. (0261-4189) 2000 brasher-2000-4 The structure of mouse HP1 suggests a unique mode of single peptide recognition by the shadow chromo domain dimer. in vitro In vitro competition binding competition binding predetermined predetermined participant predetermined Brasher SV., Smith BO., Fogh RH., Nietlispach D., Thiru A., Nielsen PR., Broadhurst RW., Ball LJ., Murzina NV., Laue ED. Purified proteins used. Mobility of the dimers on Superdex S75 gel-filteration column used to determine disruption of complex formation. 4, 5 nm@mole.bio.cam.ac.uk, e.d.laue@bioc.cam.ac.uk EMBO J. (0261-4189) 2000 cbx1_mouse Chromobox protein homolog 1 Cbx1 Cbx Heterochromatin protein 1 homolog beta Modifier 1 protein M31 Heterochromatin protein p25 protein protein mouse Mus musculus Mouse MGKKQNKKKVEEVLEEEEEEYVVEKVLDRRVVKGKVEYLLKWKGFSDEDNTWEPEENLDCPDLIAEFLQSQKTAHETDKSEGGKRKADSDSEDKGEESKPKKKKEESEKPRGFARGLEPERIIGATDSSGELMFLMKWKNSDEADLVPAKEANVKCPQVVISFYEERLTWHSYPSEDDDKKDDKN caf1a_mouse Chromatin assembly factor 1 subunit A Chromatin assembly factor I p150 subunit Chaf1a Caip150 protein protein mouse Mus musculus Mouse MLEEPEAATRTAAAVDCKDRPGFPVKRLIQARLPFKRLNLVPKEKVEEDTSPKAAVESKVPDLQLSLGTFESQCHTGSHVGLSTKLVGGQGPIDSFLRATIKPVPSVVIIDLTENCSDIPDSPEGHSELSPDTAGVVTTVEGAAKQQEHSAAELCLLETPSDITCHMEEEPGSPGDPKRTGDCQAGSLQSCPELTPGSRTCPTKELSSWSKAGDLLFIEKVPVVVLEDILATKPSIASLPMMSLDRSVTSESEILESCPEDDSILSHSSTNSSSPTSSPEGPSTPPEHRGGRSSPSTPACRVAKNFVKGSTEKGRSKLHRDREQQREEKEKLREEIRRAKEEARKKKEEEKELKEKERREKREKDEKEKAEKQRLKEEKRKERQEALEAKLEEKRKKEEEKRLREEEKRLREEEKRIKAEKAEITRFFQKPKTPQAPKTLAGSCGKFAPFEIKEHMVLAPRCRAALDQDLCDQLDQLLQQQSVASTFLSDLKSRLPLRSGPTRVCGHDTDIMNRDVVIVESSKVDGVSERKKFGRMKLLQFSENHRPAYWGTWNKKTAIIRPRNPWAQDKDLLDYEVDSDDEWEEEEPGESLSHSEGDEDDDVGEDEDEDDGFFVPHGYLSEDEGVTEECADPENHKVHQKLKAKEWDELLAKGKRFRVLQPVHVGCVWASEAANCTSSDLKLLQQFTACLLDVASPDEPEPGASRREKRDQHILAQLLPLLHGNVNGSKVIIHEFQEQCRRGLLTLPSPTPHLQMPNLEDAVAVPSKARLKRLISENSAYEKRPNFRMCWYVHPEVLKSFGQECLPVPCQWTYITTMPSAPREDSGSASTEGPGQSTPMLLKRKPAATMCITQFMKKRRYDGQVGSGDMDGFQADTEEDEEDDTDCMIIDVPDVGSDVSEAPIPAPTLCK tif1b_mouse Transcription intermediary factor 1-beta Trim28 Krip1 Tif1b Tripartite motif-containing protein 28 KRAB-A-interacting protein KRIP-1 E3 SUMO-protein ligase TRIM28 Kap1 RING-type E3 ubiquitin transferase TIF1-beta protein protein mouse Mus musculus Mouse MAASAAATAAASAATAASAASGSPGSGEGSAGGEKRPAASSAAAASAAASSPAGGGGEAQELLEHCGVCRERLRPERDPRLLPCLHSACSACLGPATPAAANNSGDGGSAGDGAMVDCPVCKQQCYSKDIVENYFMRDSGSKASSDSQDANQCCTSCEDNAPATSYCVECSEPLCETCVEAHQRVKYTKDHTVRSTGPAKTRDGERTVYCNVHKHEPLVLFCESCDTLTCRDCQLNAHKDHQYQFLEDAVRNQRKLLASLVKRLGDKHATLQKNTKEVRSSIRQVSDVQKRVQVDVKMAILQIMKELNKRGRVLVNDAQKVTEGQQERLERQHWTMTKIQKHQEHILRFASWALESDNNTALLLSKKLIYFQLHRALKMIVDPVEPHGEMKFQWDLNAWTKSAEAFGKIVAERPGTNSTGPGPMAPPRAPGPLSKQGSGSSQPMEVQEGYGFGSDDPYSSAEPHVSGMKRSRSGEGEVSGLLRKVPRVSLERLDLDLTSDSQPPVFKVFPGSTTEDYNLIVIERGAAAAAAGQAGTVPPGAPGAPPLPGMAIVKEEETEAAIGAPPAAPEGPETKPVLMPLTEGPGAEGPRLASPSGSTSSGLEVVAPEVTSAPVSGPGILDDSATICRVCQKPGDLVMCNQCEFCFHLDCHLPALQDVPGEEWSCSLCHVLPDLKEEDGSLSLDGADSTGVVAKLSPANQRKCERVLLALFCHEPCRPLHQLATDSTFSMEQPGGTLDLTLIRARLQEKLSPPYSSPQEFAQDVGRMFKQFNKLTEDKADVQSIIGLQRFFETRMNDAFGDTKFSAVLVEPPPLNLPSAGLSSQELSGPGDGP cbx1-cbx1-1 Dimer formation studied. 209643 N/A 209647 unspecified role unspecified role neutral component neutral component P83917:p.Tyr164Glu Interaction disrupted mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false chromo_shadow domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false P83917:p.Ile161Glu Interaction disrupted mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P83917:p.Ile161Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false region 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 false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Changing Tyr 164 to Leu does not disrupt dimer formation, but did decrease stability of the protein which displayed a tendency to aggregate and precipitate from solution. cbx1-chaf1a-1 Complex formation studied. 209643 N/A 209648 unspecified role unspecified role neutral component neutral component chromo-shadow domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 209647 unspecified role unspecified role neutral component neutral component chromo_shadow domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false region 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 false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 direct interaction direct interaction false false false A stoichiometry of 1:1 shown in the experiment. cbx1-cbx1-2 structure of dimer. 209644 N/A 209647 unspecified role unspecified role neutral component neutral component region 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 false chromo-shadow domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false ile161 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false tyr164 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false leu168 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false asn153 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false ala125 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false val154 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 209647 unspecified role unspecified role neutral component neutral component chromo_shadow domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false ile161 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false tyr164 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false leu168 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false ile161 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false trp170 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false gln158 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 209677 209668 209678 209669 209679 209670 209680 209671 209681 209672 209682 209673 direct interaction direct interaction false false false The background r.m.s.d. for the structure is 0.81 deg A. Stoichiometry of 1:1 shown for this interaction. cbx1-chaf1a-3 Complex formation studied by nmr. 209644 N/A 209647 unspecified role unspecified role neutral component neutral component chromo_shadow domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 209648 unspecified role unspecified role neutral component neutral component chromo-shadow domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 direct interaction direct interaction false false false A stoichiometry of 1:1 shown in the experiment. A tight complex formation is indicated. cbx1-chaf1a-2 sedimentation studied. 209645 N/A 209648 unspecified role unspecified role neutral component neutral component chromo-shadow domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 N/A 209647 unspecified role unspecified role neutral component neutral component region 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 false chromo_shadow domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 direct interaction direct interaction false false false A stoichiometry of 1:1 shown in the experiment. Ultracentrifugation gave a molecular weight of 19.17kDa for the complex which agrees with gel filteration results. cbx1-cbx1-3 sedimentation of dimer studied. 209645 N/A 209647 unspecified role unspecified role neutral component neutral component chromo_shadow domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Ultracentrifugation gave a molecular weight of 16.63 for the dimer. This agrees with the data from the gel filteration column. cbx1-chaf1a-4 competition binding between CAF and MOD1 binding to TIF studied. 209646 N/A 209649 unspecified role unspecified role bait bait hp1 box binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) N/A 209648 unspecified role unspecified role prey prey chromo-shadow domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 209647 unspecified role unspecified role prey prey chromo_shadow domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false P83917:p.Trp170Ala no binding to CAF MIR or TIF MIR mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P83917:p.Trp170Glu no binding to CAF MIR or TIF MIR mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P83917:p.Ile161Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P83917:p.Ile161Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P83917:p.Tyr164Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false in vitro In vitro direct interaction direct interaction false false false Competition studies of Cbx1 binding to T1f1 carried out with CAF MIR 13 mer peptide. I161 and Y164 are involved in dimerisation while W170 is involved in binding to MIR domains. Misfolding controls were carried out in this study using recombinant Y164 and I164E whose monomeric and folded state has previously been demonstrated.