UniProt UniProt http://www.uniprot.org nishikawa-2003-1 Nep98p is a component of the yeast spindle pole body and essential for nuclear division and fusion. 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 Nishikawa S., Terazawa Y., Nakayama T., Hirata A., Makio T., Endo T. endo@biochem.chem.nagoya-u.ac.jp Yeast two-hybrid library. J. Biol. Chem. 278 (9938-9943) 2003 Only protein-protein interactions imex curation nishikawa-2003-2 Nep98p is a component of the yeast spindle pole body and essential for nuclear division and fusion. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast crosslink cross-linking study crosslink weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Nishikawa S., Terazawa Y., Nakayama T., Hirata A., Makio T., Endo T. Cells expressing Nep98p and Jem1p-3HA were converted to spheroplasts, labeled with 35S-amino acids, and proteins were crosslinked with DSP. After extraction, a first round of immunoprecipitation with the anti-Nep98p antibodies was performed. The immunoprecipitates were treated with dithiothreitol to cleave a disulfide bond of the crosslinker and were subjected to the second round of immunoprecipitation with the anti-Nep98p antibodies or the anti-HA monoclonal antibody to confirm the interaction. endo@biochem.chem.nagoya-u.ac.jp Yeast two-hybrid library. J. Biol. Chem. 278 (9938-9943) 2003 Only protein-protein interactions imex curation jem1_yeast DnaJ-like chaperone JEM1 YJL073W J1083 HRC558 DnaJ-like protein of the ER membrane 1 JEM1 KAR8 protein protein yeast Saccharomyces cerevisiae Baker's yeast MILISGYCLLVYSVILPVLISASKLCDLAELQRLNKNLKVDTESLPKYQWIAGQLEQNCMTADPASENMSDVIQLANQIYYKIGLIQLSNDQHLRAINTFEKIVFNETYKGSFGKLAEKRLQELYVDFGMWDKVHQKDDQYAKYLSLNETIRNKISSKDVSVEEDISELLRITPYDVNVLSTHIDVLFHKLAEEIDVSLAAAIILDYETILDKHLASLSIDTRLSIHYVISVLQTFVLNSDASFNIRKCLSIDMDYDKCKKLSLTISKLNKVNPSKRQILDPATYAFENKKFRSWDRIIEFYLKDKKPFITPMKILNKDTNFKNNYFFLEEIIKQLIEDVQLSRPLAKNLFEDPPITDGFVKPKSYYHTDYLVYIDSILCQASSMSPDVKRAKLAAPFCKKSLRHSLTLETWKHYQDAKSEQKPLPETVLSDVWNSNPHLLMYMVNSILNKSRSKPHSQFKKQLYDQINKFFQDNGLSESTNPYVMKNFRLLQKQLQTYKEHKHRNFNQQYFQQQQQQQQHQRHQAPPAAPNYDPKKDYYKILGVSPSASSKEIRKAYLNLTKKYHPDKIKANHNDKQESIHETMSQINEAYETLSDDDKRKEYDLSRSNPRRNTFPQGPRQNNMFKNPGSGFPFGNGFKMNFGL mps3_yeast Spindle pole body assembly component MPS3 J1310 J1315 YJL018W YJL019W MPS3 NEP98 Monopolar spindle protein 3 98 kDa nuclear envelope protein protein protein yeast Saccharomyces cerevisiae Baker's yeast MNNSNEHRREEAGAANEQMPYNKAVKSAYADVLKDKMNREQEISLRAIKKGIYTDGGETDNYDMDKENDSAYEMFKKNLDFPLDQHNDDDDDDPYIEDNGQETDGYSDEDYTDEADKSFIEDSDSDSYDLESNSDFEENLESSGEAKKLKWRTYIFYGGLFFVFYFFGSFLMTTVKNNDLESHSSGATSSPGKSFSNLQKQVNHLYSELSKRDEKHSSELDKTVKIIVSQFEKNIKRLLPSNLVNFENDINSLTKQVETISTSMSELQRRNHKFTVENVTQWQDQLVKQLDTHLPQEIPVVINNSSSLLIIPELHNYLSALISDVIESPGIGTAGSAESRWEYDLNRYVKEILSNELQYIDKDYFIQEMNRRLQSNKQEIWEEITNRLETQQQQQQQQVQQDYSNVPQQYSSILMKRLIHQIYNSNQHQWEDDLDFATYVQGTKLLNHLTSPTWRQGSGVQPIELLTDSKQSSSTYWQCENEPGCSWAIRFKTPLYLTKISYMHGRFTNNLHIMNSAPRLISLYVKLSQTKEIKALQTLANQYGFGQHHKRDRNYIKIAKFEYRLTDSRIRQQMYLPPWFIQLKPLVRSIVFQVDENYGNKKFISLRKFIINGVTPQDLQIIENNEFPVLLGDTPEYGVTQNTDEGKRKVLLSKPPYASSSTSTKFHPASNVPSFGQDELDQ jem1-nep98 Jem1p and Nep98p interact in a yeast two-hybrid assay 681639 N/A 681641 unspecified role unspecified role bait bait N/A 681642 unspecified role unspecified role prey prey region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false No figure jem1-nep98-1 Jem1p and Nep98p interact in a cross-linking assay 681640 N/A 681642 unspecified role unspecified role neutral component neutral component N/A 681641 unspecified role unspecified role neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig. 2C