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/ The structure of mouse HP1 suggests a unique mode of single peptide recognition by the shadow chromo domain dimer. The structure of mouse HP1 suggests a unique mode of single peptide recognition by the shadow chromo domain dimer. EMBO J. (0261-4189) 2000 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 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 Annotation to indicate that this entry has been checked and accepted by a senior curator. EMBO J. (0261-4189) 2000 The structure of mouse HP1 suggests a unique mode of single peptide recognition by the shadow chromo domain dimer. The structure of mouse HP1 suggests a unique mode of single peptide recognition by the shadow chromo domain dimer. EMBO J. (0261-4189) 2000 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 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 Annotation to indicate that this entry has been checked and accepted by a senior curator. EMBO J. (0261-4189) 2000 The structure of mouse HP1 suggests a unique mode of single peptide recognition by the shadow chromo domain dimer. The structure of mouse HP1 suggests a unique mode of single peptide recognition by the shadow chromo domain dimer. EMBO J. (0261-4189) 2000 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 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 Annotation to indicate that this entry has been checked and accepted by a senior curator. EMBO J. (0261-4189) 2000 The structure of mouse HP1 suggests a unique mode of single peptide recognition by the shadow chromo domain dimer. The structure of mouse HP1 suggests a unique mode of single peptide recognition by the shadow chromo domain dimer. EMBO J. (0261-4189) 2000 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 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 Annotation to indicate that this entry has been checked and accepted by a senior curator. EMBO J. (0261-4189) 2000 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 DIP protein Q62318 original sequence version: 129 DIP protein Q62318 original sequence version: 120 DIP protein Q62318 original sequence version: 138 DIP protein Q62318 original sequence version: 152 DIP protein Q62318 original sequence version: 166 DIP protein Q62318 original sequence version: 139 DE87AAA5DC67BB8B 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 DIP protein P83917 original sequence version: 93 BE687AF9C66E48E3 caf1a_mouse Chromatin assembly factor 1 subunit A Chromatin assembly factor I p150 subunit Chaf1a Caip150 protein protein mouse Mus musculus Mouse MLEEPEAATRTAAAVDCKDRPGFPVKRLIQARLPFKRLNLVPKEKVEEDTSPKAAVESKVPDLQLSLGTFESQCHTGSHVGLSTKLVGGQGPIDSFLRATIKPVPSVVIIDLTENCSDIPDSPEGHSELSPDTAGVVTTVEGAAKQQEHSAAELCLLETPSDITCHMEEEPGSPGDPKRTGDCQAGSLQSCPELTPGSRTCPTKELSSWSKAGDLLFIEKVPVVVLEDILATKPSIASLPMMSLDRSVTSESEILESCPEDDSILSHSSTNSSSPTSSPEGPSTPPEHRGGRSSPSTPACRVAKNFVKGSTEKGRSKLHRDREQQREEKEKLREEIRRAKEEARKKKEEEKELKEKERREKREKDEKEKAEKQRLKEEKRKERQEALEAKLEEKRKKEEEKRLREEEKRLREEEKRIKAEKAEITRFFQKPKTPQAPKTLAGSCGKFAPFEIKEHMVLAPRCRAALDQDLCDQLDQLLQQQSVASTFLSDLKSRLPLRSGPTRVCGHDTDIMNRDVVIVESSKVDGVSERKKFGRMKLLQFSENHRPAYWGTWNKKTAIIRPRNPWAQDKDLLDYEVDSDDEWEEEEPGESLSHSEGDEDDDVGEDEDEDDGFFVPHGYLSEDEGVTEECADPENHKVHQKLKAKEWDELLAKGKRFRVLQPVHVGCVWASEAANCTSSDLKLLQQFTACLLDVASPDEPEPGASRREKRDQHILAQLLPLLHGNVNGSKVIIHEFQEQCRRGLLTLPSPTPHLQMPNLEDAVAVPSKARLKRLISENSAYEKRPNFRMCWYVHPEVLKSFGQECLPVPCQWTYITTMPSAPREDSGSASTEGPGQSTPMLLKRKPAATMCITQFMKKRRYDGQVGSGDMDGFQADTEEDEEDDTDCMIIDVPDVGSDVSEAPIPAPTLCK 67B3340010425C95 cbx1-cbx1-1 4 6 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 Y E Wed May 02 20:14:31 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr164glu' Tue Jun 30 18:03:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr164Glu' chromo_shadow domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain P83917:p.Ile161Glu Interaction disrupted mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain I E Wed May 02 16:36:47 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile161glu' Tue Jun 30 18:19:28 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile161Glu' P83917:p.Ile161Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain I A Wed May 02 17:11:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile161ala' Tue Jun 30 15:51:39 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile161Ala' 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction 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 4 7 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 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 ecoli Escherichia coli (strain K12) 6 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 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction A stoichiometry of 1:1 shown in the experiment. cbx1-cbx1-2 1 6 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 chromo-shadow domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ile161 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain tyr164 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain leu168 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain asn153 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ala125 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain val154 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 6 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 ile161 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain tyr164 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain leu168 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ile161 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain trp170 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain gln158 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 35 26 36 27 28 37 38 29 39 30 31 40 direct interaction direct interaction The background r.m.s.d. for the structure is 0.81 deg A. Stoichiometry of 1:1 shown for this interaction. cbx1-chaf1a-2 3 7 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 chemical synthesis Chemical synthesis (Chemical synthesis) 6 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 chromo_shadow domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction 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 3 6 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Ultracentrifugation gave a molecular weight of 16.63 for the dimer. This agrees with the data from the gel filteration column. cbx1-chaf1a-3 1 6 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 region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 7 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 chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction A stoichiometry of 1:1 shown in the experiment. A tight complex formation is indicated. cbx1-chaf1a-4 2 5 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 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 ecoli Escherichia coli (strain K12) 7 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 chemical synthesis Chemical synthesis (Chemical synthesis) 6 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 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 W A Wed May 02 16:35:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp170ala' Tue Jun 30 18:07:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp170Ala' 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 W E Wed May 02 15:08:05 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp170glu' Tue Jun 30 17:33:33 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp170Glu' P83917:p.Ile161Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain I A Wed May 02 14:25:59 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile161ala' Tue Jun 30 17:39:14 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile161Ala' P83917:p.Ile161Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain I E Wed May 02 14:33:51 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile161glu' Tue Jun 30 17:42:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile161Glu' P83917:p.Tyr164Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain Y E Wed May 02 14:40:36 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr164glu' Tue Jun 30 18:48:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr164Glu' in vitro In vitro direct interaction direct interaction 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.