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 http://mint.bio.uniroma2.it/mint patel-2006-2 yeast Saccharomyces cerevisiae Baker's yeast two hybrid pooling two hybrid pooling approach two hybrid pooling nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Patel A., Cummings N., Batchelor M., Hill PJ., Dubois T., Mellits KH., Frankel G., Connerton I. ian.connerton@nottingham.ac.uk Cellular microbiology 2006 Only protein-protein interactions imex curation imex curation patel-2006-4 in vitro In vitro elisa enzyme linked immunosorbent assay ELISA ELISA elisa antibody detection identification by antibody antibody detection Patel A., Cummings N., Batchelor M., Hill PJ., Dubois T., Mellits KH., Frankel G., Connerton I. ian.connerton@nottingham.ac.uk Cellular microbiology 2006 Only protein-protein interactions imex curation imex curation patel-2006-5 human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Patel A., Cummings N., Batchelor M., Hill PJ., Dubois T., Mellits KH., Frankel G., Connerton I. ian.connerton@nottingham.ac.uk Cellular microbiology 2006 Only protein-protein interactions imex curation imex curation patel-2006-3 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 Patel A., Cummings N., Batchelor M., Hill PJ., Dubois T., Mellits KH., Frankel G., Connerton I. ian.connerton@nottingham.ac.uk Cellular microbiology 2006 Only protein-protein interactions imex curation imex curation patel-2006-6 human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Patel A., Cummings N., Batchelor M., Hill PJ., Dubois T., Mellits KH., Frankel G., Connerton I. ian.connerton@nottingham.ac.uk Cellular microbiology 2006 Only protein-protein interactions imex curation imex curation 1433b_human 14-3-3 protein beta/alpha YWHAB Protein kinase C inhibitor protein 1 Protein 1054 protein protein human Homo sapiens Human MTMDKSELVQKAKLAEQAERYDDMAAAMKAVTEQGHELSNEERNLLSVAYKNVVGARRSSWRVISSIEQKTERNEKKQQMGKEYREKIEAELQDICNDVLELLDKYLIPNATQPESKVFYLKMKGDYFRYLSEVASGDNKQTTVSNSQQAYQEAFEISKKEMQPTHPIRLGLALNFSVFYYEILNSPEKACSLAKTAFDEAIAELDTLNEESYKDSTLIMQLLRDNLTLWTSENQGDEGDAGEGEN tir_eco27 Translocated intimin receptor Tir tir Secreted effector protein Tir espE E2348C_3941 protein protein eco27 Escherichia coli O127:H6 (strain E2348/69 / EPEC) MPIGNLGNNVNGNHLIPPAPPLPSQTDGAARGGTGHLISSTGALGSRSLFSPLRNSMADSVDSRDIPGLPTNPSRLAAATSETCLLGGFEVLHDKGPLDILNTQIGPSAFRVEVQADGTHAAIGEKNGLEVSVTLSPQEWSSLQSIDTEGKNRFVFTGGRGGSGHPMVTVASDIAEARTKILAKLDPDNHGGRQPKDVDTRSVGVGSASGIDDGVVSETHTSTTNSSVRSDPKFWVSVGAIAAGLAGLAATGIAQALALTPEPDDPTTTDPDQAANAAESATKDQLTQEAFKNPENQKVNIDANGNAIPSGELKDDIVEQIAQQAKEAGEVARQQAVESNAQAQQRYEDQHARRQEELQLSSGIGYGLSSALIVAGGIGAGVTTALHRRNQPAEQTTTTTTHTVVQQQTGGNTPAQGGTDATRAEDASLNRRDSQGSVASTHWSDSSSEVVNPYAEVGGARNSLSAHQPEEHIYDEVAADPGYSVIQNFSGSGPVTGRLIGTPGQGIQSTYALLANSGGLRLGMGGLTSGGESAVSSVNAAPTPGPVRFV 1433z_human 14-3-3 protein zeta/delta YWHAZ Protein kinase C inhibitor protein 1 protein protein human Homo sapiens Human MDKNELVQKAKLAEQAERYDDMAACMKSVTEQGAELSNEERNLLSVAYKNVVGARRSSWRVVSSIEQKTEGAEKKQQMAREYREKIETELRDICNDVLSLLEKFLIPNASQAESKVFYLKMKGDYYRYLAEVAAGDDKKGIVDQSQQAYQEAFEISKKEMQPTHPIRLGLALNFSVFYYEILNSPEKACSLAKTAFDEAIAELDTLSEESYKDSTLIMQLLRDNLTLWTSDTQGDEAEAGEGGEN 1433t_human 14-3-3 protein theta YWHAQ 14-3-3 protein tau 14-3-3 protein T-cell Protein HS1 protein protein human Homo sapiens Human MEKTELIQKAKLAEQAERYDDMATCMKAVTEQGAELSNEERNLLSVAYKNVVGGRRSAWRVISSIEQKTDTSDKKLQLIKDYREKVESELRSICTTVLELLDKYLIANATNPESKVFYLKMKGDYFRYLAEVACGDDRKQTIDNSQGAYQEAFDISKKEMQPTHPIRLGLALNFSVFYYEILNNPELACTLAKTAFDEAIAELDTLNEDSYKDSTLIMQLLRDNLTLWTSDSAGEECDAAEGAEN k1c18_human Keratin, type I cytoskeletal 18 KRT18 CYK18 Keratin-18 PIG46 Cytokeratin-18 Cell proliferation-inducing gene 46 protein protein protein human Homo sapiens Human MSFTTRSTFSTNYRSLGSVQAPSYGARPVSSAASVYAGAGGSGSRISVSRSTSFRGGMGSGGLATGIAGGLAGMGGIQNEKETMQSLNDRLASYLDRVRSLETENRRLESKIREHLEKKGPQVRDWSHYFKIIEDLRAQIFANTVDNARIVLQIDNARLAADDFRVKYETELAMRQSVENDIHGLRKVIDDTNITRLQLETEIEALKEELLFMKKNHEEEVKGLQAQIASSGLTVEVDAPKSQDLAKIMADIRAQYDELARKNREELDKYWSQQIEESTTVVTTQSAEVGAAETTLTELRRTVQSLEIDLDSMRNLKASLENSLREVEARYALQMEQLNGILLHLESELAQTRAEGQRQAQEYEALLNIKVKLEAEIATYRRLLEDGEDFNLGDALDSSNSMQTIQKTTTRRIVDGKVVSETNDTKVLRH tir-ywhab-1 3637461 N/A 3637466 unspecified role unspecified role prey prey cdna library cdna library N/A 3637467 predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed physical association physical association false false false table 1 fig. 1 tir-ywhaz-1 3637461 N/A 3637468 unspecified role unspecified role prey prey cdna library cdna library N/A 3637467 predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed physical association physical association false false false table 1 fig. 1 tir-ywhaq-1 3637461 N/A 3637469 unspecified role unspecified role prey prey cdna library cdna library N/A 3637467 predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed physical association physical association false false false table 1 fig. 1 ywhaq-tir-1 3637462 N/A 3637469 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified ecoli Escherichia coli (strain K12) N/A 3637467 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 false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false fig. 2b ywhaq-tir-2 3637462 N/A 3637466 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified ecoli Escherichia coli (strain K12) N/A 3637467 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 false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false fig. 2b By comparison at saturation of 14‐3‐3τ with Tir, the 14‐3‐3β isoform exhibited only 10% of the binding ability of the former. tir-krt18-2 3637463 N/A 3637470 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3637469 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3637467 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep infection infection association association false false false fig 3 The 14‐3‐3τ and cytokeratin 18 proteins were observed to co‐precipitate with Tir in the wild‐type EPEC‐infected membrane and cytoplasmic extracts but were absent in the ΔTir control. tir-krt18-3 3637463 N/A 3637469 unspecified role unspecified role bait bait subcellular prep subcellular preparation subcellular prep physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3637467 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep infection infection physical association physical association false false false fig 3 tir-ywhaq-2 3637464 N/A 3637469 unspecified role unspecified role prey prey cdna library cdna library in performing these experiments we noted that pGAD‐GH constructs missing the 3′ non‐coding region present in cDNA33 exhibited lower levels of β‐galactosidase activity in the yeast two‐hybrid assay irrespective of the codons at the beginning of the 14‐3‐3 reading frame (PJ69‐4A pICC10 + pNC701 and pICC10 + pNC720 of Fig. 1A). N/A 3637467 predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed B7UM99:p.Ser51Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false B7UM99:p.Ser434Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false B7UM99:p.Ser465Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false table 1 fig. 1 None of these Tir subfragments showed β‐galactosidase activities above that of the host or single plasmid‐bearing control strains (Fig. 1B). In yeast, the complete Tir molecule is required for recognition by 14‐3‐3τ. tir-ywhaq-3 3637465 N/A 3637467 unspecified role unspecified role neutral component neutral component infection infection fixed cell fixed cell N/A 3637469 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization false false false fig. 4 14‐3‐3τ was found throughout the host cell but was observed to accumulate adjacent to Tir localized at the extremity of the actin‐stained pedestal. The tips of the pedestals represent the interface of the bacterial attachment site and the location of Tir as it interacts with intimin