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/ Yng1 PHD finger binding to H3 trimethylated at K4 promotes NuA3 HAT activity at K14 of H3 and transcription at a subset of targeted ORFs. Yng1 PHD finger binding to H3 trimethylated at K4 promotes NuA3 HAT activity at K14 of H3 and transcription at a subset of targeted ORFs. Mol. Cell (1097-2765) 2006 imex curation Taverna SD., Ilin S., Rogers RS., Tanny JC., Lavender H., Li H., Baker L., Boyle J., Blair LP., Chait BT., Patel DJ., Aitchison JD., Tackett AJ., Allis CD. Chromatin - Epigenetic interactions resulting in chromatin modulation Tagged YNG1 cells, grown in isotopically light media, and wild-type (untagged) cells, grown in heavy isotopic (d4-lysine) media, were mixed in a 1:1 ratio by cell weight and colysed. Protein A (PrA) component of the TAP tag was utilized for purification using IgG-coated Dynabeads. Analysis allowed identification of proteins that were either stably associated (peptides yielding an exclusively light labeled signature) or nonstably associated (1:1 ratio of light to heavy labeled peptides) with tagged Yng1. Proteins identified as containing near 100% h4-lysine (isotopically light) were regarded as true protein complex components, while protein identifications containing 50% h4-lysine (and therefore 50% d4-lysine) aas contaminants (I-DIRT analysis). ajtackett@uams.edu, alliscd@rockefeller.edu 29-Mar-2010: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro fps fluorescence polarization spectroscopy Fluorescence anisotropy FPS FPS Fluorescence anisotropy fps predetermined predetermined participant predetermined Taverna SD., Ilin S., Rogers RS., Tanny JC., Lavender H., Li H., Baker L., Boyle J., Blair LP., Chait BT., Patel DJ., Aitchison JD., Tackett AJ., Allis CD. Chromatin - Epigenetic interactions resulting in chromatin modulation Tagged YNG1 cells, grown in isotopically light media, and wild-type (untagged) cells, grown in heavy isotopic (d4-lysine) media, were mixed in a 1:1 ratio by cell weight and colysed. Protein A (PrA) component of the TAP tag was utilized for purification using IgG-coated Dynabeads. Analysis allowed identification of proteins that were either stably associated (peptides yielding an exclusively light labeled signature) or nonstably associated (1:1 ratio of light to heavy labeled peptides) with tagged Yng1. Proteins identified as containing near 100% h4-lysine (isotopically light) were regarded as true protein complex components, while protein identifications containing 50% h4-lysine (and therefore 50% d4-lysine) aas contaminants (I-DIRT analysis). ajtackett@uams.edu, alliscd@rockefeller.edu Accepted 2010-APR-16 by JYOTI. 29-Mar-2010: Contacted by JYOTI. Mol. Cell (1097-2765) 2006 jyoti Only protein-protein interactions imex curation Yng1 PHD finger binding to H3 trimethylated at K4 promotes NuA3 HAT activity at K14 of H3 and transcription at a subset of targeted ORFs. Yng1 PHD finger binding to H3 trimethylated at K4 promotes NuA3 HAT activity at K14 of H3 and transcription at a subset of targeted ORFs. Mol. Cell (1097-2765) 2006 imex curation Taverna SD., Ilin S., Rogers RS., Tanny JC., Lavender H., Li H., Baker L., Boyle J., Blair LP., Chait BT., Patel DJ., Aitchison JD., Tackett AJ., Allis CD. Chromatin - Epigenetic interactions resulting in chromatin modulation Tagged YNG1 cells, grown in isotopically light media, and wild-type (untagged) cells, grown in heavy isotopic (d4-lysine) media, were mixed in a 1:1 ratio by cell weight and colysed. Protein A (PrA) component of the TAP tag was utilized for purification using IgG-coated Dynabeads. Analysis allowed identification of proteins that were either stably associated (peptides yielding an exclusively light labeled signature) or nonstably associated (1:1 ratio of light to heavy labeled peptides) with tagged Yng1. Proteins identified as containing near 100% h4-lysine (isotopically light) were regarded as true protein complex components, while protein identifications containing 50% h4-lysine (and therefore 50% d4-lysine) aas contaminants (I-DIRT analysis). ajtackett@uams.edu, alliscd@rockefeller.edu 29-Mar-2010: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro acetylase assay acetylase assay predetermined predetermined participant predetermined Taverna SD., Ilin S., Rogers RS., Tanny JC., Lavender H., Li H., Baker L., Boyle J., Blair LP., Chait BT., Patel DJ., Aitchison JD., Tackett AJ., Allis CD. Chromatin - Epigenetic interactions resulting in chromatin modulation Tagged YNG1 cells, grown in isotopically light media, and wild-type (untagged) cells, grown in heavy isotopic (d4-lysine) media, were mixed in a 1:1 ratio by cell weight and colysed. Protein A (PrA) component of the TAP tag was utilized for purification using IgG-coated Dynabeads. Analysis allowed identification of proteins that were either stably associated (peptides yielding an exclusively light labeled signature) or nonstably associated (1:1 ratio of light to heavy labeled peptides) with tagged Yng1. Proteins identified as containing near 100% h4-lysine (isotopically light) were regarded as true protein complex components, while protein identifications containing 50% h4-lysine (and therefore 50% d4-lysine) aas contaminants (I-DIRT analysis). ajtackett@uams.edu, alliscd@rockefeller.edu Accepted 2010-APR-16 by JYOTI. 29-Mar-2010: Contacted by JYOTI. Mol. Cell (1097-2765) 2006 jyoti Only protein-protein interactions imex curation Yng1 PHD finger binding to H3 trimethylated at K4 promotes NuA3 HAT activity at K14 of H3 and transcription at a subset of targeted ORFs. Yng1 PHD finger binding to H3 trimethylated at K4 promotes NuA3 HAT activity at K14 of H3 and transcription at a subset of targeted ORFs. Mol. Cell (1097-2765) 2006 imex curation Taverna SD., Ilin S., Rogers RS., Tanny JC., Lavender H., Li H., Baker L., Boyle J., Blair LP., Chait BT., Patel DJ., Aitchison JD., Tackett AJ., Allis CD. Chromatin - Epigenetic interactions resulting in chromatin modulation Tagged YNG1 cells, grown in isotopically light media, and wild-type (untagged) cells, grown in heavy isotopic (d4-lysine) media, were mixed in a 1:1 ratio by cell weight and colysed. Protein A (PrA) component of the TAP tag was utilized for purification using IgG-coated Dynabeads. Analysis allowed identification of proteins that were either stably associated (peptides yielding an exclusively light labeled signature) or nonstably associated (1:1 ratio of light to heavy labeled peptides) with tagged Yng1. Proteins identified as containing near 100% h4-lysine (isotopically light) were regarded as true protein complex components, while protein identifications containing 50% h4-lysine (and therefore 50% d4-lysine) aas contaminants (I-DIRT analysis). ajtackett@uams.edu, alliscd@rockefeller.edu 29-Mar-2010: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Taverna SD., Ilin S., Rogers RS., Tanny JC., Lavender H., Li H., Baker L., Boyle J., Blair LP., Chait BT., Patel DJ., Aitchison JD., Tackett AJ., Allis CD. Chromatin - Epigenetic interactions resulting in chromatin modulation Tagged YNG1 cells, grown in isotopically light media, and wild-type (untagged) cells, grown in heavy isotopic (d4-lysine) media, were mixed in a 1:1 ratio by cell weight and colysed. Protein A (PrA) component of the TAP tag was utilized for purification using IgG-coated Dynabeads. Analysis allowed identification of proteins that were either stably associated (peptides yielding an exclusively light labeled signature) or nonstably associated (1:1 ratio of light to heavy labeled peptides) with tagged Yng1. Proteins identified as containing near 100% h4-lysine (isotopically light) were regarded as true protein complex components, while protein identifications containing 50% h4-lysine (and therefore 50% d4-lysine) aas contaminants (I-DIRT analysis). ajtackett@uams.edu, alliscd@rockefeller.edu Accepted 2010-APR-16 by JYOTI. 29-Mar-2010: Contacted by JYOTI. Mol. Cell (1097-2765) 2006 jyoti Only protein-protein interactions imex curation Yng1 PHD finger binding to H3 trimethylated at K4 promotes NuA3 HAT activity at K14 of H3 and transcription at a subset of targeted ORFs. Yng1 PHD finger binding to H3 trimethylated at K4 promotes NuA3 HAT activity at K14 of H3 and transcription at a subset of targeted ORFs. Mol. Cell (1097-2765) 2006 imex curation Taverna SD., Ilin S., Rogers RS., Tanny JC., Lavender H., Li H., Baker L., Boyle J., Blair LP., Chait BT., Patel DJ., Aitchison JD., Tackett AJ., Allis CD. Chromatin - Epigenetic interactions resulting in chromatin modulation Tagged YNG1 cells, grown in isotopically light media, and wild-type (untagged) cells, grown in heavy isotopic (d4-lysine) media, were mixed in a 1:1 ratio by cell weight and colysed. Protein A (PrA) component of the TAP tag was utilized for purification using IgG-coated Dynabeads. Analysis allowed identification of proteins that were either stably associated (peptides yielding an exclusively light labeled signature) or nonstably associated (1:1 ratio of light to heavy labeled peptides) with tagged Yng1. Proteins identified as containing near 100% h4-lysine (isotopically light) were regarded as true protein complex components, while protein identifications containing 50% h4-lysine (and therefore 50% d4-lysine) aas contaminants (I-DIRT analysis). ajtackett@uams.edu, alliscd@rockefeller.edu 29-Mar-2010: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Taverna SD., Ilin S., Rogers RS., Tanny JC., Lavender H., Li H., Baker L., Boyle J., Blair LP., Chait BT., Patel DJ., Aitchison JD., Tackett AJ., Allis CD. Chromatin - Epigenetic interactions resulting in chromatin modulation Tagged YNG1 cells, grown in isotopically light media, and wild-type (untagged) cells, grown in heavy isotopic (d4-lysine) media, were mixed in a 1:1 ratio by cell weight and colysed. Protein A (PrA) component of the TAP tag was utilized for purification using IgG-coated Dynabeads. Analysis allowed identification of proteins that were either stably associated (peptides yielding an exclusively light labeled signature) or nonstably associated (1:1 ratio of light to heavy labeled peptides) with tagged Yng1. Proteins identified as containing near 100% h4-lysine (isotopically light) were regarded as true protein complex components, while protein identifications containing 50% h4-lysine (and therefore 50% d4-lysine) aas contaminants (I-DIRT analysis). ajtackett@uams.edu, alliscd@rockefeller.edu Accepted 2010-APR-16 by JYOTI. 29-Mar-2010: Contacted by JYOTI. Mol. Cell (1097-2765) 2006 jyoti Only protein-protein interactions imex curation Yng1 PHD finger binding to H3 trimethylated at K4 promotes NuA3 HAT activity at K14 of H3 and transcription at a subset of targeted ORFs. Yng1 PHD finger binding to H3 trimethylated at K4 promotes NuA3 HAT activity at K14 of H3 and transcription at a subset of targeted ORFs. Mol. Cell (1097-2765) 2006 imex curation Taverna SD., Ilin S., Rogers RS., Tanny JC., Lavender H., Li H., Baker L., Boyle J., Blair LP., Chait BT., Patel DJ., Aitchison JD., Tackett AJ., Allis CD. Chromatin - Epigenetic interactions resulting in chromatin modulation Tagged YNG1 cells, grown in isotopically light media, and wild-type (untagged) cells, grown in heavy isotopic (d4-lysine) media, were mixed in a 1:1 ratio by cell weight and colysed. Protein A (PrA) component of the TAP tag was utilized for purification using IgG-coated Dynabeads. Analysis allowed identification of proteins that were either stably associated (peptides yielding an exclusively light labeled signature) or nonstably associated (1:1 ratio of light to heavy labeled peptides) with tagged Yng1. Proteins identified as containing near 100% h4-lysine (isotopically light) were regarded as true protein complex components, while protein identifications containing 50% h4-lysine (and therefore 50% d4-lysine) aas contaminants (I-DIRT analysis). ajtackett@uams.edu, alliscd@rockefeller.edu 29-Mar-2010: Contacted by JYOTI. jyoti Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast pull down pull down sequence tag sequence tag identification sequence tag Taverna SD., Ilin S., Rogers RS., Tanny JC., Lavender H., Li H., Baker L., Boyle J., Blair LP., Chait BT., Patel DJ., Aitchison JD., Tackett AJ., Allis CD. Chromatin - Epigenetic interactions resulting in chromatin modulation Tagged YNG1 cells, grown in isotopically light media, and wild-type (untagged) cells, grown in heavy isotopic (d4-lysine) media, were mixed in a 1:1 ratio by cell weight and colysed. Protein A (PrA) component of the TAP tag was utilized for purification using IgG-coated Dynabeads. Analysis allowed identification of proteins that were either stably associated (peptides yielding an exclusively light labeled signature) or nonstably associated (1:1 ratio of light to heavy labeled peptides) with tagged Yng1. Proteins identified as containing near 100% h4-lysine (isotopically light) were regarded as true protein complex components, while protein identifications containing 50% h4-lysine (and therefore 50% d4-lysine) aas contaminants (I-DIRT analysis). ajtackett@uams.edu, alliscd@rockefeller.edu Accepted 2010-APR-16 by JYOTI. 29-Mar-2010: Contacted by JYOTI. Mol. Cell (1097-2765) 2006 jyoti Only protein-protein interactions imex curation Yng1 PHD finger binding to H3 trimethylated at K4 promotes NuA3 HAT activity at K14 of H3 and transcription at a subset of targeted ORFs. Yng1 PHD finger binding to H3 trimethylated at K4 promotes NuA3 HAT activity at K14 of H3 and transcription at a subset of targeted ORFs. Mol. Cell (1097-2765) 2006 imex curation Taverna SD., Ilin S., Rogers RS., Tanny JC., Lavender H., Li H., Baker L., Boyle J., Blair LP., Chait BT., Patel DJ., Aitchison JD., Tackett AJ., Allis CD. Chromatin - Epigenetic interactions resulting in chromatin modulation Tagged YNG1 cells, grown in isotopically light media, and wild-type (untagged) cells, grown in heavy isotopic (d4-lysine) media, were mixed in a 1:1 ratio by cell weight and colysed. Protein A (PrA) component of the TAP tag was utilized for purification using IgG-coated Dynabeads. Analysis allowed identification of proteins that were either stably associated (peptides yielding an exclusively light labeled signature) or nonstably associated (1:1 ratio of light to heavy labeled peptides) with tagged Yng1. Proteins identified as containing near 100% h4-lysine (isotopically light) were regarded as true protein complex components, while protein identifications containing 50% h4-lysine (and therefore 50% d4-lysine) aas contaminants (I-DIRT analysis). ajtackett@uams.edu, alliscd@rockefeller.edu 29-Mar-2010: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Taverna SD., Ilin S., Rogers RS., Tanny JC., Lavender H., Li H., Baker L., Boyle J., Blair LP., Chait BT., Patel DJ., Aitchison JD., Tackett AJ., Allis CD. Chromatin - Epigenetic interactions resulting in chromatin modulation Tagged YNG1 cells, grown in isotopically light media, and wild-type (untagged) cells, grown in heavy isotopic (d4-lysine) media, were mixed in a 1:1 ratio by cell weight and colysed. Protein A (PrA) component of the TAP tag was utilized for purification using IgG-coated Dynabeads. Analysis allowed identification of proteins that were either stably associated (peptides yielding an exclusively light labeled signature) or nonstably associated (1:1 ratio of light to heavy labeled peptides) with tagged Yng1. Proteins identified as containing near 100% h4-lysine (isotopically light) were regarded as true protein complex components, while protein identifications containing 50% h4-lysine (and therefore 50% d4-lysine) aas contaminants (I-DIRT analysis). ajtackett@uams.edu, alliscd@rockefeller.edu Accepted 2010-APR-16 by JYOTI. 29-Mar-2010: Contacted by JYOTI. Mol. Cell (1097-2765) 2006 jyoti Only protein-protein interactions imex curation nto1_yeast NuA3 HAT complex component NTO1 YPR031W NTO1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MNRGSLDDGPKLREEKHFQDFYPDLNADTLLPFIVPLVETKDNSTDTDSDDISNRNNREIGSVKSVQTKELIFKGRVTTEPLVLKKNEVEFQKCKITTNELKGKKNPYCVRFNESFISRYYHINKVRNRKSYKQQQKEFDGVEAPYFTKFSSKEAPNITISTSTKSAIQKFASISPNLVNFKPQYDMDEQDELYLHYLNKRYFKDQMSHEIFEILMTTLETEWFHIEKHIPSTNSLIARHNILRDCKNYELYGSDDGTGLSMDQACAVCLGTDSDNLNTIVFCDGCDIAVHQECYGIIFIPEGKWLCRRCMISKNNFATCLMCPSHTGAFKQTDTGSWVHNICALWLPELYFSNLHYMEPIEGVQNVSVSRWKLNCYICKKKMGACIQCFQRNCFTAYHVTCARRAGLYMSKGKCTIQELASNQFSQKYSVESFCHKHAPRGWQTSIEGINKARKYFSLLSTLQTETPQHNEANDRTNSKFNKTIWKTPNQTPVAPHVFAEILQKVVDFFGLANPPAGAFDICKYWSMKRELTGGTPLTACFENNSLGSLTEEQVQTRIDFANDQLEDLYRLKELTTLVKKRTQASNSLSRSRKKVFDIVKSPQKYLLKINVLDIFIKSEQFKALERLVTEPKLLVILEKCKHCDFDTVQIFKEEIMHFFEVLETLPGASRILQTVSSKAKEQVTNLIGLIEHVDIKKLLSRDFIINDDKIEERPWSGPVIMEEEGLSDAEELSAGEHRMLKLILNSG 7D6796553BCFA06D 7D6796553BCFA06D yng1_yeast Protein YNG1 ING1 homolog 1 YNG1 YOR29-15 YOR064C protein protein yeast Saccharomyces cerevisiae Baker's yeast MEHLANENSDSDIRYSFLSTLDHLPCELIRSLRLMQTIDLFKNEEDEPGMERACRDLLLVATYINDLVDDQIHFLKQHKKELEIQKSVTKNFNSSLENIKSKLTLEEPGAYKEPKLLLKINLKKAKSRERKESITSPTIGINQGDVTEGNNNQEEVYCFCRNVSYGPMVACDNPACPFEWFHYGCVGLKQAPKGKWYCSKDCKEIANQRSKSKRQKRRK 5232A71839E2FDA4 5232A71839E2FDA4 eaf6_yeast Chromatin modification-related protein EAF6 YJR082C J1854 ESA1-associated factor 6 EAF6 protein protein yeast Saccharomyces cerevisiae Baker's yeast MTDELKSYEALKAELKKSLQDRREQEDTFDNLQQEIYDKETEYFSHNSNNNHSGHGGAHGSKSHYSGNIIKGFDTFSKSHHSHADSAFNNNDRIFSLSSATYVKQQHGQSQND 396029BD06F31E6C h3_yeast Histone H3 SIN2 YNL031C N2749 HHT1 YBR010W YBR0201 HHT2 protein protein yeast Saccharomyces cerevisiae Baker's yeast MARTKQTARKSTGGKAPRKQLASKAARKSAPSTGGVKKPHRYKPGTVALREIRRFQKSTELLIRKLPFQRLVREIAQDFKTDLRFQSSAIGALQESVEAYLVSLFEDTNLAAIHAKRVTIQKKDIKLARRLRGERS A6115FEB480AC67A A6115FEB480AC67A taf14_yeast Transcription initiation factor TFIID subunit 14 ANC1 CST10 YPL129W TAF14 SWP29 TAF30 TFG3 TBP-associated factor 14 TBP-associated factor 30 kDa Transcription initiation factor TFIIF 30 kDa subunit Transcription factor G 30 kDa subunit SWI/SNF chromatin-remodeling complex subunit TAF14 SWI/SNF complex subunit TAF14 SWI/SNF complex 29 kDa subunit Actin non-complementing mutant 1 Chromosome stability protein 10 protein protein yeast Saccharomyces cerevisiae Baker's yeast MVATVKRTIRIKTQQHILPEVPPVENFPVRQWSIEIVLLDDEGKEIPATIFDKVIYHLHPTFANPNRTFTDPPFRIEEQGWGGFPLDISVFLLEKAGERKIPHDLNFLQESYEVEHVIQIPLNKPLLTEELAKSGSTEETTANTGTIGKRRTTTNTTAEPKAKRAKTGSASTVKGSVDLEKLAFGLTKLNEDDLVGVVQMVTDNKTPEMNVTNNVEEGEFIIDLYSLPEGLLKSLWDYVKKNTE 60EA5EC2474B1B9C 60EA5EC2474B1B9C sas3_yeast Histone acetyltransferase SAS3 SAS3 YBL052C YBL0515 YBL0507 Something about silencing protein 3 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSLTANDESPKPKKNALLKNLEIDDLIHSQFVRSDTNGHRTTRRLFNSDASISHRIRGSVRSDKGLNKIKKGLISQQSKLASENSSQNIVNRDNKMGAVSFPIIEPNIEVSEELKVRIKYDSIKFFNFERLISKSSVIAPLVNKNITSSGPLIGFQRRVNRLKQTWDLATENMEYPYSSDNTPFRDNDSWQWYVPYGGTIKKMKDFSTKRTLPTWEDKIKFLTFLENSKSATYINGNVSLCNHNETDQENEDRKKRKGKVPRIKNKVWFSQIEYIVLRNYEIKPWYTSPFPEHINQNKMVFICEFCLKYMTSRYTFYRHQLKCLTFKPPGNEIYRDGKLSVWEIDGRENVLYCQNLCLLAKCFINSKTLYYDVEPFIFYILTEREDTENHPYQNAAKFHFVGYFSKEKFNSNDYNLSCILTLPIYQRKGYGQFLMEFSYLLSRKESKFGTPEKPLSDLGLLTYRTFWKIKCAEVLLKLRDSARRRSNNKNEDTFQQVSLNDIAKLTGMIPTDVVFGLEQLQVLYRHKTRSLSSLDDFNYIIKIDSWNRIENIYKTWSSKNYPRVKYDKLLWEPIILGPSFGINGMMNLEPTALADEALTNETMAPVISNNTHIENYNNSRAHNKRRRRRRRSSEHKTSKLHVNNIIEPEVPATDFFEDTVSSLTEYMCDYKNTNNDRLIYQAEKRVLESIHDRKGIPRSKFSTETHWELCFTIKNSETPLGNHAARRNDTGISSLEQDEVENDVDTELYVGENAKEDEDEDEDFTLDDDIEDEQISEENDEEEDTYEEDSDDDEDGKRKGQEQDENDIESHIRKERVRKRRKITLIEDDEE ACF5B1B225CB4A71 yng1-eaf6-1 5 9 unspecified role unspecified role prey prey 11 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 12 unspecified role unspecified role prey prey 8 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 We inferred from the I-DIRT analysis that the stable NuA3 complex contains five proteins: Sas3 (98 kDa), Nto1 (86 kDa), Taf30 (27 kDa), Yng1 (25 kDa), and Eaf6 (13 kDa) (Figure 1B) Fig. 1B h3-yng1-1 3 8 unspecified role unspecified role prey prey Q08465:p.Trp180Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain W E Wed May 02 17:41:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp180glu' Tue Jun 30 18:52:57 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp180Glu' region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 10 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain lys-5 (required) n6me3+lys N6,N6,N6-trimethyl-L-lysine (5S)-5-amino-5-carboxy-N,N,N-trimethylpentan-1-aminium 2-amino-6-(trimethylammonio)hexanoic acid 5-azanyl-5-carboxy-N,N,N-trimethylpentanazanium MOD_RES N6,N6,N6-trimethyllysine N(zeta)-trimethyllysine N6,N6,N6-trimethyl-L-lysine N6,N6,N6-trimethylated L-lysine N6,N6,N6-trimethyllysin-N6-ium N6,N6,N6-trimethyllysine cation epsilon-trimethyllysine N-trimethylation (of lysine) N6Me3+Lys Trimethyl tri-Methylation certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association Fig. 2C Interaction observed in nto1 and sas3 -/- cells h3-yng1-2 3 9 unspecified role unspecified role prey prey region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role prey prey region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 11 unspecified role unspecified role prey prey region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 10 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain lys-5 (required) n6me3+lys N6,N6,N6-trimethyl-L-lysine (5S)-5-amino-5-carboxy-N,N,N-trimethylpentan-1-aminium 2-amino-6-(trimethylammonio)hexanoic acid 5-azanyl-5-carboxy-N,N,N-trimethylpentanazanium MOD_RES N6,N6,N6-trimethyllysine N(zeta)-trimethyllysine N6,N6,N6-trimethyl-L-lysine N6,N6,N6-trimethylated L-lysine N6,N6,N6-trimethyllysin-N6-ium N6,N6,N6-trimethyllysine cation epsilon-trimethyllysine N-trimethylation (of lysine) N6Me3+Lys Trimethyl tri-Methylation certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 8 unspecified role unspecified role prey prey region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 12 unspecified role unspecified role prey prey region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Fig. 2A Although this experiment was done as a series of bait/prey pairs, subsequent knockout experiments showed only Yng1 interacts directly with the peptide so the data has been represented as a one bait many prey, to show a truer representation. Low levels of binding were observed with the dimethylated peptide. h3-yng1-3 4 10 unspecified role unspecified role prey prey lys-5 (required) n6me3+lys N6,N6,N6-trimethyl-L-lysine (5S)-5-amino-5-carboxy-N,N,N-trimethylpentan-1-aminium 2-amino-6-(trimethylammonio)hexanoic acid 5-azanyl-5-carboxy-N,N,N-trimethylpentanazanium MOD_RES N6,N6,N6-trimethyllysine N(zeta)-trimethyllysine N6,N6,N6-trimethyl-L-lysine N6,N6,N6-trimethylated L-lysine N6,N6,N6-trimethyllysin-N6-ium N6,N6,N6-trimethyllysine cation epsilon-trimethyllysine N-trimethylation (of lysine) N6Me3+Lys Trimethyl tri-Methylation certain certain sequence position certain certain certain sequence position certain yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 8 unspecified role unspecified role bait bait n-terminal 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 Q08465:p.Trp180Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain W E Wed May 02 20:20:19 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp180glu' Tue Jun 30 20:05:35 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp180Glu' phd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 3A Yng1 Binds Directly to H3 K4me3 through Its Single PHD Finger A lower level of binding was observed to dimethylated Histone H3. h3-yng1-4 6 10 unspecified role unspecified role neutral component neutral component lys-5 (required) n6me3+lys N6,N6,N6-trimethyl-L-lysine (5S)-5-amino-5-carboxy-N,N,N-trimethylpentan-1-aminium 2-amino-6-(trimethylammonio)hexanoic acid 5-azanyl-5-carboxy-N,N,N-trimethylpentanazanium MOD_RES N6,N6,N6-trimethyllysine N(zeta)-trimethyllysine N6,N6,N6-trimethyl-L-lysine N6,N6,N6-trimethylated L-lysine N6,N6,N6-trimethyllysin-N6-ium N6,N6,N6-trimethyllysine cation epsilon-trimethyllysine N-trimethylation (of lysine) N6Me3+Lys Trimethyl tri-Methylation certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 8 unspecified role unspecified role neutral component neutral component region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined phd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 1H isotope label isotope label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 56 53 direct interaction direct interaction Figs. 3B & 4 Supp. Fig. 2 Yng1 Binds Directly to H3 K4me3 through Its Single PHD Finger A lower level of binding was observed to dimethylated Histone H3. h3-yng1-5 1 8 unspecified role unspecified role neutral component neutral component region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined phd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 1H isotope label isotope label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 10 unspecified role unspecified role neutral component neutral component lys-5 (required) n6me3+lys N6,N6,N6-trimethyl-L-lysine (5S)-5-amino-5-carboxy-N,N,N-trimethylpentan-1-aminium 2-amino-6-(trimethylammonio)hexanoic acid 5-azanyl-5-carboxy-N,N,N-trimethylpentanazanium MOD_RES N6,N6,N6-trimethyllysine N(zeta)-trimethyllysine N6,N6,N6-trimethyl-L-lysine N6,N6,N6-trimethylated L-lysine N6,N6,N6-trimethyllysin-N6-ium N6,N6,N6-trimethyllysine cation epsilon-trimethyllysine N-trimethylation (of lysine) N6Me3+Lys Trimethyl tri-Methylation certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Figs. 3C, 3D Yng1 Binds Directly to H3 K4me3 through Its Single PHD Finger KD H3K5(Me)3=9.1E-6M, KD H3K5(Me)2=2.1E-5M, KD H3K5(Me)1=5.1E-5M Temp= room temperature, pH 7.5. yng1-eaf6-2 2 10 enzyme target enzyme target substrate substrate neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain lys-5 n6me3+lys N6,N6,N6-trimethyl-L-lysine (5S)-5-amino-5-carboxy-N,N,N-trimethylpentan-1-aminium 2-amino-6-(trimethylammonio)hexanoic acid 5-azanyl-5-carboxy-N,N,N-trimethylpentanazanium MOD_RES N6,N6,N6-trimethyllysine N(zeta)-trimethyllysine N6,N6,N6-trimethyl-L-lysine N6,N6,N6-trimethylated L-lysine N6,N6,N6-trimethyllysin-N6-ium N6,N6,N6-trimethyllysine cation epsilon-trimethyllysine N-trimethylation (of lysine) N6Me3+Lys Trimethyl tri-Methylation certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 8 unspecified role unspecified role ancillary ancillary 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 12 enzyme enzyme neutral component neutral component yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 11 unspecified role unspecified role ancillary ancillary yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 7 unspecified role unspecified role ancillary ancillary yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 9 unspecified role unspecified role ancillary ancillary yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast acetylation acetylation reaction acetylation NuA3 exhibits enhanced HAT activity upon trimethylation of H3K4. Fig. 5