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 Src-dependent autophagic degradation of Ret in FAK-signalling-defective cancer cells. Src-dependent autophagic degradation of Ret in FAK-signalling-defective cancer cells. EMBO Rep. (1469-221X) 2012 imex curation Sandilands E., Serrels B., Wilkinson S., Frame MC. e.sandilands@ed.ac.uk Only protein-protein interactions in vitro In vitro pull down pull down experimental particp experimental participant identification experimental particp Sandilands E., Serrels B., Wilkinson S., Frame MC. e.sandilands@ed.ac.uk EMBO Rep. (1469-221X) 2012 Only protein-protein interactions imex curation Src-dependent autophagic degradation of Ret in FAK-signalling-defective cancer cells. Src-dependent autophagic degradation of Ret in FAK-signalling-defective cancer cells. EMBO Rep. (1469-221X) 2012 imex curation Sandilands E., Serrels B., Wilkinson S., Frame MC. e.sandilands@ed.ac.uk Only protein-protein interactions mouse-scc-1 Mus musculus scc-1 scc-1 fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Sandilands E., Serrels B., Wilkinson S., Frame MC. e.sandilands@ed.ac.uk EMBO Rep. (1469-221X) 2012 Only protein-protein interactions imex curation Src-dependent autophagic degradation of Ret in FAK-signalling-defective cancer cells. Src-dependent autophagic degradation of Ret in FAK-signalling-defective cancer cells. EMBO Rep. (1469-221X) 2012 imex curation Sandilands E., Serrels B., Wilkinson S., Frame MC. e.sandilands@ed.ac.uk Only protein-protein interactions mouse-scc-1 Mus musculus scc-1 scc-1 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Sandilands E., Serrels B., Wilkinson S., Frame MC. e.sandilands@ed.ac.uk EMBO Rep. (1469-221X) 2012 Only protein-protein interactions imex curation src_mouse Neuronal proto-oncogene tyrosine-protein kinase Src Src pp60c-src Proto-oncogene c-Src protein protein mouse Mus musculus Mouse MGSNKSKPKDASQRRRSLEPSENVHGAGGAFPASQTPSKPASADGHRGPSAAFVPPAAEPKLFGGFNSSDTVTSPQRAGPLAGGVTTFVALYDYESRTETDLSFKKGERLQIVNNTEGDWWLAHSLSTGQTGYIPSNYVAPSDSIQAEEWYFGKITRRESERLLLNAENPRGTFLVRESETTKGAYCLSVSDFDNAKGLNVKHYKIRKLDSGGFYITSRTQFNSLQQLVAYYSKHADGLCHRLTTVCPTSKPQTQGLAKDAWEIPRESLRLEVKLGQGCFGEVWMGTWNGTTRVAIKTLKPGTMSPEAFLQEAQVMKKLRHEKLVQLYAVVSEEPIYIVTEYMNKGSLLDFLKGETGKYLRLPQLVDMSAQIASGMAYVERMNYVHRDLRAANILVGENLVCKVADFGLARLIEDNEYTARQGAKFPIKWTAPEAALYGRFTIKSDVWSFGILLTELTTKGRVPYPGMVNREVLDQVERGYRMPCPPECPESLHDLMCQCWRKEPEERPTFEYLQAFLEDYFTSTEPQYQPGENL DIP protein P05480 original sequence version: 163 DIP protein P05480 original sequence version: 99 DIP protein P05480 original sequence version: 128 DIP protein P05480 original sequence version: 136 DIP protein P05480 original sequence version: 176 22CB5C6BE7A061C0 paxi_mouse Paxillin Pxn protein protein mouse Mus musculus Mouse MDDLDALLADLESTTSHISKRPVFLSEEPPYSYPTGNHTYQEIAVPPPVPPPPSSEALNGTVLDPLDQWQPSGSRYAHQQPPSPLPVYSSSAKNSSASNTQDGVGSLCSRAGEEEHVYSFPNKQKSAEPSPTVMSSSLGSNLSELDRLLLELNAVQHSPPGFPADEAESSPPLPGALSPLYGIPENNTPLGGKAGPLVKEKPKRNGGRGLEDVRPSVESLLDELESSVPSPVPAITVNQGEMSSPQRVTSSQQQTRISASSATRELDELMASLSDFKMQGLEQRVDGERPWAAGWPPSSRQSSPEGQDEGGFMAQGKTGSSSPPGGLSKPGSQLDSMLGSLQSDLNKLGVATVAKGVCGACKKPIAGQVVTAMGKTWHPEHFVCTHCQEEIGSRNFFERDGQPYCEKDYHSLFSPRCYYCNGPILDKVVTALDRTWHPEHFFCAQCGAFFGPEGFHEKDGKAYCRKDYFDMFAPKCGGCARAILENYISALNTLWHPECFVCRECFTPFVNGSFFEHDGQPYCEVHYHERRGSLCSGCQKPITGRCITAMAKKFHPEHFVCAFCLKQLNKGTFKEQNDKPYCQSCFVKLFC B41A1892E6F60544 ret_mouse Proto-oncogene tyrosine-protein kinase receptor Ret Ret Proto-oncogene c-Ret protein protein mouse Mus musculus Mouse MAKATSGAAGLGLKLILLLPLLGEAPLGLYFSRDAYWERLYVDQPAGTPLLYVHALRDAPGEVPSFRLGQHLYGVYRTRLHENDWIRINETTGLLYLNQSLDHSSWEQLSIRNGGFPLLTIFLQVFLGSTAQREGECHWPGCTRVYFSFINDTFPNCSSFKAQDLCIPETAVSFRVRENRPPGTFYHFHMLPVQFLCPNISVKYSLLGGDSLPFRCDPDCLEVSTRWALDRELREKYVLEALCIVAGPGANKETVTLSFPVTVYDEDDSAPTFSGGVGTASAVVEFKRKEGTVVATLQVFDADVVPASGELVRRYTNTLLSGDSWAQQTFRVEHSPIETLVQVNNNSVRATMHNYKLILNRSLSISESRVLQLAVLVNDSDFQGPGAGGILVLHFNVSVLPVTLNLPRAYSFPVNKRARRYAQIGKVCVENCQEFSGVSIQYKLQPSSINCTALGVVTSPEDTSGTLFVNDTEALRRPECTKLQYTVVATDRQTRRQTQASLVVTVEGTSITEEVGCPKSCAVNKRRPECEECGGLGSPTGRCEWRQGDGKGITRNFSTCSPSTRTCPDGHCDAVESRDANICPQDCLRADIVGGHERGERQGIKAGYGICNCFPDEKKCFCEPEDSQGPLCDALCRTIITAALFSLIISILLSIFCVCHHHKHGHKPPIASAEMTFCRPAQGFPISYSSSGTRRPSLDSTENQVPVDSFKIPEDPKWEFPRKNLVLGKTLGEGEFGKVVKATAFRLKGRAGYTTVAVKMLKENASQSELRDLLSEFNLLKQVNHPHVIKLYGACSQDGPLLLIVEYAKYGSLRGFLRDSRKIGPAYVSGGGSRNSSSLDHPDERVLTMGDLISFAWQISRGMQYLAEMKLVHRDLAARNILVAEGRKMKISDFGLSRDVYEEDSYVKKSKGRIPVKWMAIESLFDHIYTTQSDVWSFGVLLWEIVTLGGNPYPGIPPERLFNLLKTGHRMERPDNCSEEMYRLMLQCWKQEPDKRPVFADISKDLEKMMVKSRDYLDLAASTPSDSLLYDDGLSEEETPLVDCNNAPLPRSLPSTWIENKLYGMSDPNWPGESPVPLTRADGTSTGFPRYANDSVYANWMVSPSAAKLMDTFDS A5CF1EF45A640413 mlp3b_mouse Microtubule-associated proteins 1A/1B light chain 3B Map1lc3b Map1alc3 Map1lc3 Microtubule-associated protein 1 light chain 3 beta MAP1A/MAP1B light chain 3 B MAP1 light chain 3-like protein 2 Autophagy-related protein LC3 B Autophagy-related ubiquitin-like modifier LC3 B protein protein mouse Mus musculus Mouse MPSEKTFKQRRSFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLPVLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESERDEDGFLYMVYASQETFGTAMAV DIP protein Q9CQV6 original sequence version: 118 DIP protein Q9CQV6 original sequence version: 123 mint Probably involved in formation of autophagosomal vacuoles (autophagosomes) 520E29028163FA8D ret-src-1 2 Ret 6 immunostaining immunostaining experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell Src 4 immunostaining immunostaining experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization mint f2a f3b Interestingly, we detected almost complete co-localization of Ret with activated Src tyrosine kinase (PY416) in adhesions in FAKþ/þ cells (Fig 2A, upper panels, solid arrow) and in puncta in FAK / cells (Fig 2A, lower panels, broken arrows). We have previously identified indistinguishable Src-containing puncta as autophagosomes [10], using specific antibodies against autophagy-related proteins (Atg) such as LC3B, which co-localizes with Src in adhesions (Fig 2B, upper panel, solid arrow) and in autophagosomes (Fig 2B, lower panel, broken arrows) in the presence and absence of FAK, respectively. map1lc3b-src-2 2 LC3 B 7 immunostaining immunostaining experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell Src 4 immunostaining immunostaining experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization mint Interestingly, we detected almost complete co-localization of Ret with activated Src tyrosine kinase (PY416) in adhesions in FAKþ/þ cells (Fig 2A, upper panels, solid arrow) and in puncta in FAK / cells (Fig 2A, lower panels, broken arrows). We have previously identified indistinguishable Src-containing puncta as autophagosomes [10], using specific antibodies against autophagy-related proteins (Atg) such as LC3B, which co-localizes with Src in adhesions (Fig 2B, upper panel, solid arrow) and in autophagosomes (Fig 2B, lower panel, broken arrows) in the presence and absence of FAK, respectively. f2b pxn-ret-1 2 Paxillin 5 immunostaining immunostaining experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell Ret 6 immunostaining immunostaining experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell colocalization colocalization mint When these cells were treated with dasatinib, we also observed a striking relocalization of Ret from autophagosomes to paxillin-containing focal adhesions (Fig 4C, solid arrows, quantified in the right panel). To confirm that these Ret-positive structures were autophagosomes, we treated the cells with 3-methyladenine (3MA), an inhibitor that prevents the formation of autophagosomes [20]. In FAKþ/þ cells 3MA had no effect on the localization of Ret (supplementary Fig S3a online), but in FAK / cells Ret was restored to adhesions containing paxillin (supplementary Fig S3a online, quantified in supplementary Fig S3b online), and the lower molecular weight 80-kDa Ret species was diminished (supplementary Fig S3c online). f4b f4c sf3a sf3b map1lc3b-ret 1 LC3 B 7 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp 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 Ret 6 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association mint sf4d src-ret 3 Src 4 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Ret 6 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association f1a mint