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 yeast N(alpha)-acetyltransferase NatA is quantitatively anchored to the ribosome and interacts with nascent polypeptides. The yeast N(alpha)-acetyltransferase NatA is quantitatively anchored to the ribosome and interacts with nascent polypeptides. Mol. Cell. Biol. (0270-7306) 2003 imex curation Gautschi M., Just S., Mun A., Ross S., Rucknagel P., Dubaquie Y., Ehrenhofer-Murray A., Rospert S. Sabine.Rospert@biochemie.uni-freiburg.de Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast crosslink cross-linking study crosslink weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Gautschi M., Just S., Mun A., Ross S., Rucknagel P., Dubaquie Y., Ehrenhofer-Murray A., Rospert S. Performed with low-salt treated RNCs carrying 35S-labeled prepro alpha 87 as a nascent chain. Aliquots of the cross-link reactions were subjected to immunoprecipitation under denaturing conditions. Rospert@biochemie.uni-freiburg.de Sabine.Rospert@biochemie.uni-freiburg.de Accepted 2009-MAY-1 by ORCHARD. Mol. Cell. Biol. (0270-7306) 2003 orchard Only protein-protein interactions imex curation The yeast N(alpha)-acetyltransferase NatA is quantitatively anchored to the ribosome and interacts with nascent polypeptides. The yeast N(alpha)-acetyltransferase NatA is quantitatively anchored to the ribosome and interacts with nascent polypeptides. Mol. Cell. Biol. (0270-7306) 2003 imex curation Gautschi M., Just S., Mun A., Ross S., Rucknagel P., Dubaquie Y., Ehrenhofer-Murray A., Rospert S. Sabine.Rospert@biochemie.uni-freiburg.de Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Gautschi M., Just S., Mun A., Ross S., Rucknagel P., Dubaquie Y., Ehrenhofer-Murray A., Rospert S. Rospert@biochemie.uni-freiburg.de Sabine.Rospert@biochemie.uni-freiburg.de Accepted 2009-MAY-1 by ORCHARD. Mol. Cell. Biol. (0270-7306) 2003 Only protein-protein interactions imex curation The yeast N(alpha)-acetyltransferase NatA is quantitatively anchored to the ribosome and interacts with nascent polypeptides. The yeast N(alpha)-acetyltransferase NatA is quantitatively anchored to the ribosome and interacts with nascent polypeptides. Mol. Cell. Biol. (0270-7306) 2003 imex curation Gautschi M., Just S., Mun A., Ross S., Rucknagel P., Dubaquie Y., Ehrenhofer-Murray A., Rospert S. Sabine.Rospert@biochemie.uni-freiburg.de Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast crosslink cross-linking study crosslink weight by comassie molecular weight estimation by coomasie staining weight by comassie Gautschi M., Just S., Mun A., Ross S., Rucknagel P., Dubaquie Y., Ehrenhofer-Murray A., Rospert S. Crosslinking agent was Bis(Sulfosuccinimidyl) suberate (BS3), reaction was performed in the presence of high-salt ribosome-nascent chain complexes. Rospert@biochemie.uni-freiburg.de Sabine.Rospert@biochemie.uni-freiburg.de Accepted 2009-MAY-1 by ORCHARD. Mol. Cell. Biol. (0270-7306) 2003 orchard Only protein-protein interactions imex curation The yeast N(alpha)-acetyltransferase NatA is quantitatively anchored to the ribosome and interacts with nascent polypeptides. The yeast N(alpha)-acetyltransferase NatA is quantitatively anchored to the ribosome and interacts with nascent polypeptides. Mol. Cell. Biol. (0270-7306) 2003 imex curation Gautschi M., Just S., Mun A., Ross S., Rucknagel P., Dubaquie Y., Ehrenhofer-Murray A., Rospert S. Sabine.Rospert@biochemie.uni-freiburg.de Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast crosslink cross-linking study crosslink autoradiography autoradiography Gautschi M., Just S., Mun A., Ross S., Rucknagel P., Dubaquie Y., Ehrenhofer-Murray A., Rospert S. Crosslinking reagent Bis(Sulfosuccinimidyl) suberate (BS3), performed with low-salt-treated ribosome-nascent chain complexes. Aliquots of the cross-link reactions were subjected to immunoprecipitation under denaturing conditions. Rospert@biochemie.uni-freiburg.de Sabine.Rospert@biochemie.uni-freiburg.de Accepted 2009-MAY-1 by ORCHARD. Mol. Cell. Biol. (0270-7306) 2003 orchard Only protein-protein interactions imex curation The yeast N(alpha)-acetyltransferase NatA is quantitatively anchored to the ribosome and interacts with nascent polypeptides. The yeast N(alpha)-acetyltransferase NatA is quantitatively anchored to the ribosome and interacts with nascent polypeptides. Mol. Cell. Biol. (0270-7306) 2003 imex curation Gautschi M., Just S., Mun A., Ross S., Rucknagel P., Dubaquie Y., Ehrenhofer-Murray A., Rospert S. Sabine.Rospert@biochemie.uni-freiburg.de Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast cosedimentation cosedimentation western blot western blot Immuno blot Immuno blot Gautschi M., Just S., Mun A., Ross S., Rucknagel P., Dubaquie Y., Ehrenhofer-Murray A., Rospert S. Aliquots were separated on 10% Tris-Tricine gels. Ard1p and Nat5p were initially identified by N-terminal Edman degradation, Nat1p by mass spectrometry of a tryptic digest. Rospert@biochemie.uni-freiburg.de Sabine.Rospert@biochemie.uni-freiburg.de Accepted 2009-MAY-1 by ORCHARD. Mol. Cell. Biol. (0270-7306) 2003 orchard Only protein-protein interactions imex curation nat1_yeast N-terminal acetyltransferase A complex subunit NAT1 NAT1 AAA1 YDL040C D2720 Amino-terminal, alpha-amino, acetyltransferase 1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSRKRSTKPKPAAKIALKKENDQFLEALKLYEGKQYKKSLKLLDAILKKDGSHVDSLALKGLDLYSVGEKDDAASYVANAIRKIEGASASPICCHVLGIYMRNTKEYKESIKWFTAALNNGSTNKQIYRDLATLQSQIGDFKNALVSRKKYWEAFLGYRANWTSLAVAQDVNGERQQAINTLSQFEKLAEGKISDSEKYEHSECLMYKNDIMYKAASDNQDKLQNVLKHLNDIEPCVFDKFGLLERKATIYMKLGQLKDASIVYRTLIKRNPDNFKYYKLLEVSLGIQGDNKLKKALYGKLEQFYPRCEPPKFIPLTFLQDKEELSKKLREYVLPQLERGVPATFSNVKPLYQRRKSKVSPLLEKIVLDYLSGLDPTQDPIPFIWTNYYLSQHFLFLKDFPKAQEYIDAALDHTPTLVEFYILKARILKHLGLMDTAAGILEEGRQLDLQDRFINCKTVKYFLRANNIDKAVEVASLFTKNDDSVNGIKDLHLVEASWFIVEQAEAYYRLYLDRKKKLDDLASLKKEVESDKSEQIANDIKENQWLVRKYKGLALKRFNAIPKFYKQFEDDQLDFHSYCMRKGTPRAYLEMLEWGKALYTKPMYVRAMKEASKLYFQMHDDRLKRKSDSLDENSDEIQNNGQNSSSQKKKAKKEAAAMNKRKETEAKSVAAYPSDQDNDVFGEKLIETSTPMEDFATEFYNNYSMQVREDERDYILDFEFNYRIGKLALCFASLNKFAKRFGTTSGLFGSMAIVLLHATRNDTPFDPILKKVVTKSLEKEYSENFPLNEISNNSFDWLNFYQEKFGKNDINGLLFLYRYRDDVPIGSSNLKEMIISSLSPLEPHSQNEILQYYL DCFB870D049F91E6 DCFB870D049F91E6 rl4a_yeast 60S ribosomal protein L4-A RPL4A RPL2 RPL2A YBR031W YBR0315 L2 YL2 RP2 Large ribosomal subunit protein uL4-A protein protein yeast Saccharomyces cerevisiae Baker's yeast MSRPQVTVHSLTGEATANALPLPAVFSAPIRPDIVHTVFTSVNKNKRQAYAVSEKAGHQTSAESWGTGRAVARIPRVGGGGTGRSGQGAFGNMCRGGRMFAPTKTWRKWNVKVNHNEKRYATASAIAATAVASLVLARGHRVEKIPEIPLVVSTDLESIQKTKEAVAALKAVGAHSDLLKVLKSKKLRAGKGKYRNRRWTQRRGPLVVYAEDNGIVKALRNVPGVETANVASLNLLQLAPGAHLGRFVIWTEAAFTKLDQVWGSETVASSKVGYTLPSHIISTSDVTRIINSSEIQSAIRPAGQATQKRTHVLKKNPLKNKQVLLRLNPYAKVFAAEKLGSKKAEKTGTKPAAVFTETLKHD 37D0942A10AAE89F 37D0942A10AAE89F ard1_yeast N-terminal acetyltransferase A complex catalytic subunit ARD1 ARD1 Arrest-defective protein 1 YHR013C protein protein yeast Saccharomyces cerevisiae Baker's yeast MPINIRRATINDIICMQNANLHNLPENYMMKYYMYHILSWPEASFVATTTTLDCEDSDEQDENDKLELTLDGTNDGRTIKLDPTYLAPGEKLVGYVLVKMNDDPDQQNEPPNGHITSLSVMRTYRRMGIAENLMRQALFALREVHQAEYVSLHVRQSNRAALHLYRDTLAFEVLSIEKSYYQDGEDAYAMKKVLKLEELQISNFTHRRLKENEEKLEDDLESDLLEDIIKQGVNDIIV AEB02BA5012D1137 AEB02BA5012D1137 nat5_yeast N-alpha-acetyltransferase NAT5 NAT5 YOR253W protein protein yeast Saccharomyces cerevisiae Baker's yeast MGRDICTLDNVYANNLGMLTKLAHVTVPNLYQDAFFSALFAEDSLVAKNKKPSSKKDVHFTQMAYYSEIPVGGLVAKLVPKKQNELSLKGIQIEFLGVLPNYRHKSIGSKLLKFAEDKCSECHQHNVFVYLPAVDDLTKQWFIAHGFEQVGETVNNFIKGVNGDEQDAILLKKHIS 4F09DC597A690BA0 mdhm_yeast Malate dehydrogenase, mitochondrial YKL085W MDH1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MLSRVAKRAFSSTVANPYKVTVLGAGGGIGQPLSLLLKLNHKVTDLRLYDLKGAKGVATDLSHIPTNSVVKGFTPEEPDGLNNALKDTDMVLIPAGVPRKPGMTRDDLFAINASIVRDLAAATAESAPNAAILVISNPVNSTVPIVAQVLKNKGVYNPKKLFGVTTLDSIRAARFISEVENTDPTQERVNVIGGHSGITIIPLISQTNHKLMSDDKRHELIHRIQFGGDEVVKAKNGAGSATLSMAHAGAKFANAVLSGFKGERDVIEPSFVDSPLFKSEGIEFFASPVTLGPDGIEKIHPIGELSSEEEEMLQKCKETLKKNIEKGVNFVASK 8645C4CDFB9857C8 8645C4CDFB9857C8 ssbp1_yeast Single-stranded nucleic acid-binding protein YHL034C SBP1 SSBR1 SSB1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSAEIEEATNAVNNLSINDSEQQPRAPTHKTVIDPEDTIFIGNVAHECTEDDLKQLFVEEFGDEVSVEIPIKEHTDGHIPASKHALVKFPTKIDFDNIKENYDTKVVKDREIHIKRARTPGQMQRGGFRGRGGFRGRGGFRGGFRGGYRGGFRGRGNFRGRGGARGGFNGQKREKIPLDQMERSKDTLYINNVPFKATKEEVAEFFGTDADSISLPMRKMRDQHTGRIFTSDSANRGMAFVTFSGENVDIEAKAEEFKGKVFGDRELTVDVAVIRPENDEEEIEQETGSEEKQE FF740B206DDE5EC8 FF740B206DDE5EC8 mfal1_yeast Mating factor alpha-1 YPL187W MF(ALPHA)1 MF-ALPHA-1 MFAL1 Alpha-1 mating pheromone protein protein yeast Saccharomyces cerevisiae Baker's yeast MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYLDLEGDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLDKREAEAWHWLQLKPGQPMYKREAEAEAWHWLQLKPGQPMYKREADAEAWHWLQLKPGQPMYKREADAEAWHWLQLKPGQPMY 826ACB932DE3D128 nat1-mfal1 1 6 unspecified role unspecified role bait bait 12 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 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 direct interaction direct interaction Fig 1c The 120-kDa cross-link product was quantitatively immunoprecipitated with anti-nat1 under denaturing conditions. nat1-ard1-1 2 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait association association Fig 2a Anti-nat1 antibodies immunoprecipitated Nat1p and coimmunoprecipitated Ard1p and Nat5p, with efficiencies of 90 to 100% under native conditions (Fig. 2A). ard1-nat1-3 2 6 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey association association Fig 2a The immunoprecipitation efficiency of Ard1p with anti-ard1 was approximately 70%, and Nat1p and Nat5p were coimmunoprecipitated with similar efficiency (Fig. 2A). rpl4a-nat1-1 4 6 unspecified role unspecified role prey prey region cross linker cross linker crosslinker crosslinker undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 7 unspecified role unspecified role bait bait P10664:p.Ser2Lys S2K mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain S K Wed May 02 20:33:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser2lys' Tue Jun 30 17:20:39 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser2Lys' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast direct interaction direct interaction Fig 4a Rpl4A-S(2)K efficiently formed a cross-link to Nat1p (Fig. 4A). mdh1-nat1 4 10 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction Figure 4c Nat1p only formed a cross-link to an Mdh1p longer than 100 amino acids (Fig. 4C). mdh1-ssb1 4 11 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction Figure 4c NAC and Ssb1/2p showed interaction with an Mdh1p of at least 45 amino acids. nat1-ard1-nat5 3 6 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component direct interaction direct interaction Figure 1b The finding that Nat1p, Ard1p, and Nat5p were present in equimolar amounts [...] suggests that the complex is trimeric. nat1-nat5-ard1 5 9 unspecified role unspecified role neutral component neutral component 6 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization Figure 2b At higher potassium acetate concentrations, all three subunits were simultaneously released from the ribosomal fraction (Fig. 2B). The result suggests that the bulk of NatA is associated with ribosomes at physiological salt concentrations.