MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Novel interactions at the essential N-terminus of poly(A) polymerase that could regulate poly(A) addition in Saccharomyces cerevisiae. Novel interactions at the essential N-terminus of poly(A) polymerase that could regulate poly(A) addition in Saccharomyces cerevisiae. FEBS Lett. (0014-5793) 2012 Ezeokonkwo C., Ghazy MA., Zhelkovsky A., Yeh PC., Moore C. suggested by lperfetto in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Ezeokonkwo C., Ghazy MA., Zhelkovsky A., Yeh PC., Moore C. suggested by lperfetto FEBS Lett. (0014-5793) 2012 Novel interactions at the essential N-terminus of poly(A) polymerase that could regulate poly(A) addition in Saccharomyces cerevisiae. Novel interactions at the essential N-terminus of poly(A) polymerase that could regulate poly(A) addition in Saccharomyces cerevisiae. FEBS Lett. (0014-5793) 2012 Ezeokonkwo C., Ghazy MA., Zhelkovsky A., Yeh PC., Moore C. suggested by lperfetto yeast Saccharomyces cerevisiae 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 predetermined predetermined participant predetermined Ezeokonkwo C., Ghazy MA., Zhelkovsky A., Yeh PC., Moore C. suggested by lperfetto FEBS Lett. (0014-5793) 2012 Novel interactions at the essential N-terminus of poly(A) polymerase that could regulate poly(A) addition in Saccharomyces cerevisiae. Novel interactions at the essential N-terminus of poly(A) polymerase that could regulate poly(A) addition in Saccharomyces cerevisiae. FEBS Lett. (0014-5793) 2012 Ezeokonkwo C., Ghazy MA., Zhelkovsky A., Yeh PC., Moore C. suggested by lperfetto in vitro In vitro pull down pull down predetermined predetermined participant predetermined Ezeokonkwo C., Ghazy MA., Zhelkovsky A., Yeh PC., Moore C. suggested by lperfetto FEBS Lett. (0014-5793) 2012 nab6_yeast RNA-binding protein NAB6 YML117W YM8339.02 NAB6 Nucleic acid-binding protein 6 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSNSNSKKPVANYAYRQQQDYNGMNAMVGNPMMYHPVDFVNGAGQYGPSQHPAYYTNSPLPNIPPTPFDTAYGASLFPSHLLMGSPFVSSPNMQSGYNSARSSNLKRKAYSRPVSNHNGYNGNSNSNQNNTNNGMVTPSNYYRMGRNSFSRNNNSTRNVTHNNNKGCDTRNNSGRRTFARNNIFDDILPEMLLQRPFCINYKVLPTGDDAYRTRSLLIENVDHSIDLHSIVKNFVKSNTLESAYLIEGGKSDDSKDVETKNLSILISFLTKGDCLNFYNNILQRLSEFKTFLKSEALNLKFVCLNYDPKCLPTFIESEALTENAEEADITNGSTMISASLHHNIANKDATRSIIIEFKSPVEKSDLFKKKLQFLDRSKNKRYILESIDLVNTDVPSNQFPENYAVLTFLNISMAIEVLDYLKKYSKNLGISKCFYVSLAPLVVSSARSSVANIYEGKTSTHRLSVPSVTAGNNNDSNNNGNNNKSNMSGITTLNNNSSIGVSVYGHSNMSLTSLSSSVSLNEEIDMLATKLQGVELDGTYLEINYRDYQTPTIEEHSTHLSNVKISKTTENSRQFSQDIPSPLPLNEHMFMNDSNQSNGAIIPQQLIATPSPVSPNLQMNQRVLPNPITQSLEQNFNVSAKVASSMGSDIGNRTIYIGNINPRSKAEDICNVVRGGILQSIKYIPEKKICFVTFIEAPSAVQFYANSFIDPIVLHGNMLRVGWGHYSGPLPKLISLAVTIGASRNVYVSLPEFAFKEKFIHDPQYKKLHETLSLPDAEQLREDFSTYGDIEQINYLSDSHCCWINFMNISSAISLVEEMNKESTVQNESGEVTLKRATEEKFGGRYKGLLINYGKDRCGNINKNLIAGKNSRFYKKVKRPSYNIRLSKLEEKRRQNEIDEKEKAFDKPLNLESLGISLDAHKDNGGGETGTANNTGHENESELEAENENGNETGSFGGLGLAVASSDVKRATSDETDYEDIFNKSSGSSDSSSDVEVIMHSPSDPEYALKSQTLRSSSQTVINSKRPVKIEDEEEAVGMSQLNYRSSLRQAPPRAPSTLSYNHSKNNETPMQDIFTNGETANNRKKKRGSFARHRTIPGSDVMAQYLAQVQHSTFMYAANILGASAEDNTHPDE 47B0FF7C5B98A053 47B0FF7C5B98A053 pap_yeast Poly(A) polymerase YKR002W PAP1 Polynucleotide adenylyltransferase protein protein yeast Saccharomyces cerevisiae Baker's yeast MSSQKVFGITGPVSTVGATAAENKLNDSLIQELKKEGSFETEQETANRVQVLKILQELAQRFVYEVSKKKNMSDGMARDAGGKIFTYGSYRLGVHGPGSDIDTLVVVPKHVTREDFFTVFDSLLRERKELDEIAPVPDAFVPIIKIKFSGISIDLICARLDQPQVPLSLTLSDKNLLRNLDEKDLRALNGTRVTDEILELVPKPNVFRIALRAIKLWAQRRAVYANIFGFPGGVAWAMLVARICQLYPNACSAVILNRFFIILSEWNWPQPVILKPIEDGPLQVRVWNPKIYAQDRSHRMPVITPAYPSMCATHNITESTKKVILQEFVRGVQITNDIFSNKKSWANLFEKNDFFFRYKFYLEITAYTRGSDEQHLKWSGLVESKVRLLVMKLEVLAGIKIAHPFTKPFESSYCCPTEDDYEMIQDKYGSHKTETALNALKLVTDENKEEESIKDAPKAYLSTMYIGLDFNIENKKEKVDIHIPCTEFVNLCRSFNEDYGDHKVFNLALRFVKGYDLPDEVFDENEKRPSKKSKRKNLDARHETVKRSKSDAASGDNINGTTAAVDVN 759DE5210DC8D881 759DE5210DC8D881 cft1_yeast Protein CFT1 CFT1 Cleavage factor two protein 1 YHH1 YDR301W protein protein yeast Saccharomyces cerevisiae Baker's yeast MNVYDDVLDATVVSHSLATHFTTSDYEELLVVRTNILSVYRPTRDGKLYLTDEFKFHGLITDIGLIPQKDSPLSCLLLCTGVAKISILKFNTLTNSIDTLSLHYYEGKFKGKSLVELAKISTLRMDPGSSCALLFNNDIIAFLPFHVNKNDDDEEEEDEDENIDDSELIHSMNQKSQGTNTFNKRKRTKLGDKFTAPSVVLVASELYEGAKNIIDIQFLKNFTKPTIALLYQPKLVWAGNTTISKLPTQYVILTLNIQPAESATKIESTTIAFVKELPWDLHTIVPVSNGAIIVGTNELAFLDNTGVLQSTVLLNSFADKELQKTKIINNSSLEIMFREKNTTSIWIPSSKSKNGGSNNDETLLLMDLKSNIYYIQMEAEGRLLIKFDIFKLPIVNDLLKENSNPKCITRLNATNSNKNMDLFIGFGSGNALVLRLNNLKSTIETREAHNPSSGTNSLMDINDDDDEEMDDLYADEAPENGLTTNDSKGTVETVQPFDIELLSSLRNVGPITSLTVGKVSSIDDVVKGLPNPNKNEYSLVATSGNGSGSHLTVIQTSVQPEIELALKFISITQIWNLKIKGRDRYLITTDSTKSRSDIYESDNNFKLHKGGRLRRDATTVYISMFGEEKRIIQVTTNHLYLYDTHFRRLTTIKFDYEVIHVSVMDPYILVTVSRGDIKIFELEEKNKRKLLKVDLPEILNEMVITSGLILKSNMCNEFLIGLSKSQEEQLLFTFVTADNQIIFFTKDHNDRIFQLNGVDQLNESLYISTYQLGDEIVPDPSIKQVMINKLGHDNKEEYLTILTFGGEIYQYRKLPQRRSRFYRNVTRNDLAITGAPDNAYAKGVSSIERIMHYFPDYNGYSVIFVTGSVPYILIKEDDSTPKIFKFGNIPLVSVTPWSERSVMCVDDIKNARVYTLTTDNMYYGNKLPLKQIKISNVLDDYKTLQKLVYHERAQLFLVSYCKRVPYEALGEDGEKVIGYDENVPHAEGFQSGILLINPKSWKVIDKIDFPKNSVVNEMRSSMIQINSKTKRKREYIIAGVANATTEDTPPTGAFHIYDVIEVVPEPGKPDTNYKLKEIFQEEVSGTVSTVCEVSGRFMISQSQKVLVRDIQEDNSVIPVAFLDIPVFVTDSKSFGNLLIIGDAMQGFQFIGFDAEPYRMISLGRSMSKFQTMSLEFLVNGGDMYFAATDADRNVHVLKYAPDEPNSLSGQRLVHCSSFTLHSTNSCMMLLPRNEEFGSPQVPSFQNVGGQVDGSVFKIVPLSEEKYRRLYVIQQQIIDRELQLGGLNPRMERLANDFYQMGHSMRPMLDFNVIRRFCGLAIDRRKSIAQKAGRHAHFEAWRDIINIEFSMRSLCQGK BC50D3770E9E758E BC50D3770E9E758E fip1_yeast Pre-mRNA polyadenylation factor FIP1 J1911 YJR093C FIP1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSSSEDEDDKFLYGSDSELALPSSKRSRDDEADAGASSNPDIVKRQKFDSPVEETPATARDDRSDEDIYSDSSDDDSDSDLEVIISLGPDPTRLDAKLLDSYSTAATSSSKDVISVATDVSNTITKTSDERLITEGEANQGVTATTVKATESDGNVPKAMTGSIDLDKEGIFDSVGITTIDPEVLKEKPWRQPGANLSDYFNYGFNEFTWMEYLHRQEKLQQDYNPRRILMGLLSLQQQGKLNSANDTDSNLGNIIDNNNNVNNANMSNLNSNMGNSMSGTPNPPAPPMHPSFPPLPMFGSFPPFPMPGMMPPMNQQPNQNQNQNSK 471CA2B0CDF99D0A 471CA2B0CDF99D0A pta1_yeast Pre-tRNA-processing protein PTA1 PTA1 FUN39 YAL043C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSSAEMEQLLQAKTLAMHNNPTEMLPKVLETTASMYHNGNLSKLKLPLAKFFTQLVLDVVSMDSPIANTERPFIAAQYLPLLLAMAQSTADVLVYKNIVLIMCASYPLVLDLVAKTSNQEMFDQLCMLKKFVLSHWRTAYPLRATVDDETDVEQWLAQIDQNIGVKLATIKFISEVVLSQTKSPSGNEINSSTIPDNHPVLNKPALESEAKRLLDMLLNYLIEEQYMVSSVFIGIINSLSFVIKRRPQTTIRILSGLLRFNVDAKFPLEGKSDLNYKLSKRFVERAYKNFVQFGLKNQIITKSLSSGSGSSIYSKLTKISQTLHVIGEETKSKGILNFDPSKGNSKKTLSRQDKLKYISLWKRQLSALLSTLGVSTKTPTPVSAPATGSSTENMLDQLKILQKYTLNKASHQGNTFFNNSPKPISNTYSSVYSLMNSSNSNQDVTQLPNDILIKLSTEAILQMDSTKLITGLSIVASRYTDLMNTYINSVPSSSSSKRKSDDDDDGNDNEEVGNDGPTANSKKIKMETEPLAEEPEEPEDDDRMQKMLQEEESAQEISGDANKSTSAIKEIAPPFEPDSLTQDEKLKYLSKLTKKLFELSGRQDTTRAKSSSSSSILLDDDDSSSWLHVLIRLVTRGIEAQEASDLIREELLGFFIQDFEQRVSLIIEWLNEEWFFQTSLHQDPSNYKKWSLRVLESLGPFLENKHRRFFIRLMSELPSLQSDHLEALKPICLDPARSSLGFQTLKFLIMFRPPVQDTVRDLLHQLKQEDEGLHKQCDSLLDRLK E2AB1F468B7BC8D0 E2AB1F468B7BC8D0 sub1_yeast RNA polymerase II transcriptional coactivator SUB1 SUB1 TSP1 YMR039C YM9532.04C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSYYNRYRNKRRSDNGGGNLSNSNNNNGGMPSGLSASDAIFDLGKNKRVTVRQFRNINLIDIREYYLDSSTGEMKPGKKGISLTEDLYDELLKHRLNIDEALRRLGSKRPKTKMVRLLSDDEYEDDNNNDSTNNDKDKNGKDKNSPKKRREDKSKASNESHDLEPRSKKKKPAPPTLLPHEENIQNAEREANATLIIPGQAGRKQQEERKQKEKEEAEEAKAKAVAEQEKEAKAKEKIAEPEPEPVPTLQAKKEDIVSNINESKDANSSDEEFAQSLEAEMNKAEDDISEEE 1E278C3EF2D569C8 pap1-pta1 2 PAP1 5 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain PTA1 8 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association f1b mint pap1-fip1-7 2 Fip1 7 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined PAP1 5 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f1b homomint mint fip1-pap1-1 2 Fip1 7 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined PAP1 5 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain physical association physical association f4b homomint mint nab6-pap1 2 PAP1 5 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Nab6 4 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain c-terminal c-terminal position c-term carboxy-terminus c-terminus c-term c-terminus carboxy-terminus c-terminal physical association physical association f4b mint pta1-pap1-1 3 PTA1 8 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified gst tag 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 binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain PAP1 5 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified his tag 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 binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction f2a mint We also assessed physical contact using recombinant GST-Pta1 and His6- tagged Pap1 or deltaN-Pap1. Pap1 interacted with full-length Pta1 and the 1-300 amino acid fragment of Pta1, but not with the 1-275 fragment of Pta1 (Fig. 2A). deltaN-Pap1 did not interact with any forms of Pta1, but bound to Fip1 (Fig. 2B), indicating that unfolding of the protein does not explain a lack of interaction. These findings extend the two-hybrid results by showing a direct interaction between Pap1 and Pta1 mediated by amino acids 1-300 of Pta1 and the N-terminus of Pap1 pap1-fip1-8 3 PAP1 5 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified gst tag 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 binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Fip1 7 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction f2b homomint mint deltaN-Pap1 did not interact with any forms of Pta1, but bound to Fip1 (Fig. 2B). cft1-pap1-1 3 PAP1 5 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified his tag 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 binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Cft1 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified gst tag 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 direct interaction direct interaction f3 homomint mint Thus, like its mammalian homologue, Cft1 interacts with Pap1, but in yeast, this interaction is localized to the Pap1 N-terminus. pap1-nab6 3 PAP1 5 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain gst tag 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 Nab6 4 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction f4a mint Sub1 and Nab6 interact with the Pap1 N-terminus. pap1-sub1 3 PAP1 5 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified gst tag 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 binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Sub1 9 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified tandem tag tandem tag tandem tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction f4a homomint mint Sub1 and Nab6 interact with the Pap1 N-terminus. To find other proteins that interact with the Pap1 N-terminus, a column coupled to a peptide comprising the first 18 amino acids of Pap1 was used to pull down proteins from yeast nuclear extract. Sub1, a protein implicated in both transcription and 3\' end processing [20], and references therein), was one of the proteins identified from mass spectroscopy analysis of bound proteins. Sub1 was not found on a control column using a peptide from a different yeast protein, Npl3. Pull-downs showed that Sub1 interacts directly with full-length Pap1-GST, and this interaction is lost in the absence of the Pap1 N-terminus (Fig. 4A). pap1-fip1-9 1 Fip1 7 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain PAP1 5 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified his tag 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 binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction f2b homomint mint deltaN-Pap1 did not interact with any forms of Pta1, but bound to Fip1 (Fig. 2B).