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 DSCR1 interacts with FMRP and is required for spine morphogenesis and local protein synthesis. DSCR1 interacts with FMRP and is required for spine morphogenesis and local protein synthesis. EMBO J. (0261-4189) 2012 imex curation Wang W., Zhu JZ., Chang KT., Min KT. Only protein-protein interactions human-hek-10 Homo sapiens hek-10 hek-10 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Wang W., Zhu JZ., Chang KT., Min KT. EMBO J. (0261-4189) 2012 Only protein-protein interactions imex curation DSCR1 interacts with FMRP and is required for spine morphogenesis and local protein synthesis. DSCR1 interacts with FMRP and is required for spine morphogenesis and local protein synthesis. EMBO J. (0261-4189) 2012 imex curation Wang W., Zhu JZ., Chang KT., Min KT. Only protein-protein interactions mouse-brain_hippocampal_neuron Mus musculus, brain, hippocampal neurons brain-hippocampus_neuron Hippocampal neuron fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret predetermined predetermined participant predetermined Wang W., Zhu JZ., Chang KT., Min KT. EMBO J. (0261-4189) 2012 Only protein-protein interactions imex curation DSCR1 interacts with FMRP and is required for spine morphogenesis and local protein synthesis. DSCR1 interacts with FMRP and is required for spine morphogenesis and local protein synthesis. EMBO J. (0261-4189) 2012 imex curation Wang W., Zhu JZ., Chang KT., Min KT. Only protein-protein interactions human-hek-10 Homo sapiens hek-10 hek-10 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Wang W., Zhu JZ., Chang KT., Min KT. EMBO J. (0261-4189) 2012 Only protein-protein interactions imex curation DSCR1 interacts with FMRP and is required for spine morphogenesis and local protein synthesis. DSCR1 interacts with FMRP and is required for spine morphogenesis and local protein synthesis. EMBO J. (0261-4189) 2012 imex curation Wang W., Zhu JZ., Chang KT., Min KT. Only protein-protein interactions human-hek-10 Homo sapiens hek-10 hek-10 fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Wang W., Zhu JZ., Chang KT., Min KT. EMBO J. (0261-4189) 2012 Only protein-protein interactions imex curation fmr1_mouse Synaptic functional regulator FMR1 Fmr1 Fmr-1 Fragile X mental retardation protein 1 homolog protein protein mouse Mus musculus Mouse MEELVVEVRGSNGAFYKAFVKDVHEDSITVAFENNWQPERQIPFHDVRFPPPVGYNKDINESDEVEVYSRANEKEPCCWWLAKVRMIKGEFYVIEYAACDATYNEIVTIERLRSVNPNKPATKDTFHKIKLEVPEDLRQMCAKESAHKDFKKAVGAFSVTYDPENYQLVILSINEVTSKRAHMLIDMHFRSLRTKLSLILRNEEASKQLESSRQLASRFHEQFIVREDLMGLAIGTHGANIQQARKVPGVTAIDLDEDTCTFHIYGEDQDAVKKARSFLEFAEDVIQVPRNLVGKVIGKNGKLIQEIVDKSGVVRVRIEAENEKSVPQEEEIMPPSSLPSNNSRVGPNSSEEKKHLDTKENTHFSQPNSTKVQRVLVVSSIVAGGPQKPEPKAWQGMVPFVFVGTKDSIANATVLLDYHLNYLKEVDQLRLERLQIDEQLRQIGASSRPPPNRTDKEKGYVTDDGQGMGRGSRPYRNRGHGRRGPGYTSGTNSEASNASETESDHRDELSDWSLAPTEEERESFLRRGDGRRRRGGGRGQGGRGRGGGFKGNDDHSRTDNRPRNPREAKGRTADGSLQSASSEGSRLRTGKDRNQKKEKPDSVDGLQPLVNGVP 093DD90D589ED066 rcan1_mouse Calcipressin-1 Regulator of calcineurin 1 Down syndrome critical region protein 1 homolog Myocyte-enriched calcineurin-interacting protein 1 Rcan1 Dscr1 protein protein mouse Mus musculus Mouse MEDGVAGPRLGEVAEAVEARAPRRVTLRPFAPFSAAAEGDGGGGGDWSFIDCEMEEVDLQDLPSATIACHLDPRVFVDGLCRAKFESLFRTYDKDTTFQYFKSFKRVRINFSNPLSAADARLRLHKTEFLGKEMKLYFAQTLHIGSSHLAPPNPDKQFLISPPASPPVGWKQVEDATPVINYDLLYAISKLGPGEKYELHAATDTTPSVVVHVCESDQENEEEEEEMERMKRPKPKIIQTRRPEYTPIHLS 09F47C73D847FB2B fmr1_human Synaptic functional regulator FMR1 FMR1 Fragile X mental retardation protein 1 protein protein human Homo sapiens Human MEELVVEVRGSNGAFYKAFVKDVHEDSITVAFENNWQPDRQIPFHDVRFPPPVGYNKDINESDEVEVYSRANEKEPCCWWLAKVRMIKGEFYVIEYAACDATYNEIVTIERLRSVNPNKPATKDTFHKIKLDVPEDLRQMCAKEAAHKDFKKAVGAFSVTYDPENYQLVILSINEVTSKRAHMLIDMHFRSLRTKLSLIMRNEEASKQLESSRQLASRFHEQFIVREDLMGLAIGTHGANIQQARKVPGVTAIDLDEDTCTFHIYGEDQDAVKKARSFLEFAEDVIQVPRNLVGKVIGKNGKLIQEIVDKSGVVRVRIEAENEKNVPQEEEIMPPNSLPSNNSRVGPNAPEEKKHLDIKENSTHFSQPNSTKVQRVLVASSVVAGESQKPELKAWQGMVPFVFVGTKDSIANATVLLDYHLNYLKEVDQLRLERLQIDEQLRQIGASSRPPPNRTDKEKSYVTDDGQGMGRGSRPYRNRGHGRRGPGYTSGTNSEASNASETESDHRDELSDWSLAPTEEERESFLRRGDGRRRGGGGRGQGGRGRGGGFKGNDDHSRTDNRPRNPREAKGRTTDGSLQIRVDCNNERSVHTKTLQNTSSEGSRLRTGKDRNQKKEKPDSVDGQQPLVNGVP DIP protein Q06787 original sequence version: 68 F853D6C82E3489B9 fmr1-rcan1 1 FMRP 7 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 DSCR1 6 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f4a homomint mint To examine whether DSCR1 binds to FMRP, we immunoprecipitated FMRP with an FMRP-specific antibody and checked for the presence of DSCR1 via western blot with a DSCR1-specific antibody. We found no interaction when the immunoprecipitation was performed with the FMRP antibody (Figure 4A). Since FMRP is also present as phosphorylated protein in neurons (Ceman et al, 2003; Narayanan et al, 2007), we next checked to see if DSCR1 can interact with phospho-FMRP. Figure 4A shows that DSCR1 is brought down by an antibody specific for phospho-FMRP. Note that we were not able to detect DSCR1 interaction when performing immunoprecipitation with the FMRP antibody because the FMRP antibody preferentially binds to non-phosphorylated FMRP, and with significantly less binding to phosphorylated FMRP (Supplementary Figures S8A and B). fmr1-rcan1-1 3 DSCR1 6 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell FMRP 5 western blot western blot Immuno blot Immuno blot experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tandem tag tandem tag tandem tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P35922:p.Ser499Ala mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S A Tue Jul 12 10:21:21 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 19:02:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser499ala' Tue Jun 30 17:00:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser499Ala' physical association physical association homomint mint Next, to verify the interaction between DSCR1 and phospho-FMRP, we mutated the phosphorylation site on FMRP (serine residue at 499 to alanine). HEK cells transfected with either Flag- His-FMRP or Flag-His-FMRP(S499 to A) were used to perform immunoprecipitation with antibody against the Flag tag. We found that in cells transfected with Flag-His-FMRP, phosphorylated FMRP and DSCR1 were detected in the immuoprecipitates, but not in cells transfected with Flag-His- FMRP(S499 to A) (Figure 4B; Supplementary Figure S8C). f4b sf8c fmr1-rcan1-2 4 DSCR1 6 tag fluorescence tag visualisation by fluorescence tag fluorescence experimental particp experimental participant identification experimental particp electron donor electron donor neutral component neutral component over-expressed over expressed level over-expressed living cell living cell gfp tag gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined FMRP 5 tag fluorescence tag visualisation by fluorescence tag fluorescence experimental particp experimental participant identification experimental particp electron acceptor electron acceptor neutral component neutral component 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 P35922:p.Ser499Ala mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S A Tue Jul 12 10:18:27 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 16:44:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser499ala' Tue Jun 30 19:40:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser499Ala' proximity proximity close-contact colocalization neighbourhood interaction f4d homomint mint fmr1-rcan1-3 2 FMRP 5 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp electron donor electron donor neutral component neutral component living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level DSCR1 6 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp electron acceptor electron acceptor neutral component neutral component living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level proximity proximity close-contact colocalization neighbourhood interaction f4c homomint mint We used phospho-FMRP antibody detected with secondary antibody conjugated with Alexa 488 as donor and DSCR1 antibody detected with a secondary antibody conjugated with Alexa 555 as acceptor. Following photo-bleaching of the FRET acceptor, protein interaction is indicated by an increase in donor fluorescence. We found strong FRET signals when DSCR1 and phospho-FMRP are colocalized in dendritic spines.