MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url&gt;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 xu-2018-3 human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Xu Z., Duan F., Lu H., Abdulkadhim Dragh M., Xia Y., Liang H., Hong L. lhong@mail.hust.edu.cn <lhong@mail.hust.edu.cn>; Cell death & disease 2018 imex curation Xu Z., Duan F., Lu H., Abdulkadhim Dragh M., Xia Y., Liang H., Hong L., Xu Z., Duan F., Lu H., Abdulkadhim Dragh M., Xia Y., Liang H., Hong L. Only protein-protein interactions imex curation xu-2018-1 human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. fluorescence imaging fluorescence microscopy fluorescence imaging tag fluorescence tag visualisation by fluorescence tag fluorescence Xu Z., Duan F., Lu H., Abdulkadhim Dragh M., Xia Y., Liang H., Hong L. lhong@mail.hust.edu.cn <lhong@mail.hust.edu.cn>; Cell death & disease 2018 imex curation Xu Z., Duan F., Lu H., Abdulkadhim Dragh M., Xia Y., Liang H., Hong L., Xu Z., Duan F., Lu H., Abdulkadhim Dragh M., Xia Y., Liang H., Hong L. Only protein-protein interactions imex curation xu-2018-2 human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Xu Z., Duan F., Lu H., Abdulkadhim Dragh M., Xia Y., Liang H., Hong L. lhong@mail.hust.edu.cn <lhong@mail.hust.edu.cn>; Cell death & disease 2018 imex curation Xu Z., Duan F., Lu H., Abdulkadhim Dragh M., Xia Y., Liang H., Hong L., Xu Z., Duan F., Lu H., Abdulkadhim Dragh M., Xia Y., Liang H., Hong L. Only protein-protein interactions imex curation xu-2018-5 human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. bifc bimolecular fluorescence complementation bifc tag fluorescence tag visualisation by fluorescence tag fluorescence Xu Z., Duan F., Lu H., Abdulkadhim Dragh M., Xia Y., Liang H., Hong L. lhong@mail.hust.edu.cn <lhong@mail.hust.edu.cn>; Cell death & disease 2018 imex curation Xu Z., Duan F., Lu H., Abdulkadhim Dragh M., Xia Y., Liang H., Hong L., Xu Z., Duan F., Lu H., Abdulkadhim Dragh M., Xia Y., Liang H., Hong L. Only protein-protein interactions imex curation rash_human GTPase HRas HRAS HRAS1 Transforming protein p21 p21ras H-Ras-1 c-H-ras Ha-Ras protein protein human Homo sapiens Human MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVVIDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVKDSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREIRQHKLRKLNPPDESGPGCMSCKCVLS ubia1_human UbiA prenyltransferase domain-containing protein 1 UBIAD1 TERE1 Transitional epithelial response protein 1 protein protein human Homo sapiens Human MAASQVLGEKINILSGETVKAGDRDPLGNDCPEQDRLPQRSWRQKCASYVLALRPWSFSASLTPVALGSALAYRSHGVLDPRLLVGCAVAVLAVHGAGNLVNTYYDFSKGIDHKKSDDRTLVDRILEPQDVVRFGVFLYTLGCVCAACLYYLSPLKLEHLALIYFGGLSGSFLYTGGIGFKYVALGDLIILITFGPLAVMFAYAIQVGSLAIFPLVYAIPLALSTEAILHSNNTRDMESDREAGIVTLAILIGPTFSYILYNTLLFLPYLVFSILATHCTISLALPLLTIPMAFSLERQFRSQAFNKLPQRTAKLNLLLGLFYVFGIILAPAGSLPKI hras-ubiad1-4 3531023 N/A 3531027 unspecified role unspecified role prey prey 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 false N/A 3531028 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false fig. 4c, fig. 3d 2-bromopalmitate ubiad1-hras-1 3531023 N/A 3531027 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P01112:p.Gly12Val mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false N/A 3531028 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell physical association physical association false false false suppl fig. 3d immunoprecipitation of UBIAD1 could pull-down H-Ras (Fig. 4c), and vice versa (Supplementary Fig. S3d, e). hras-ubiad1-5 3531023 N/A 3531027 unspecified role unspecified role bait bait 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 false binding region binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false N/A 3531028 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false fig. 5d hras-ubiad1-6 3531023 N/A 3531027 unspecified role unspecified role prey prey 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 false binding region binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false N/A 3531028 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false fig. 5e, fig.5c, suppl. fig.4d immunoprecipitation of UBIAD1 could pull-down H-Ras (Fig. 4c), and vice versa (Supplementary Fig. S3d, e). To determine the binding region of H-Ras for UBIAD1, a truncated H-Ras protein was fused with GFP. As shown in Fig. 5d and Supplementary Fig. S4d, the C-terminus of H-Ras interacted with UBIAD1. hras-ubiad1-1 3531024 N/A 3531027 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell rfp tag rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3531028 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed 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 false Q9Y5Z9:p.Asn102Ser mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false Q9Y5Z9:p.Arg54_Ser57delinsAlaAlaAlaAla mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false colocalization colocalization false false false fig.2a; suppl fig 2b, fig.7a The results showed that UBIAD1 increased the retention of H-Ras in the Golgi apparatus after treatment with cycloheximide, which blocks protein synthesis (Fig. 2a). Same colocalization was detected in T24 cell. hras-ubiad1-2 3531024 N/A 3531027 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell bfp tag fluorescent prot tag fluorescent protein tag fluorescent prot tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3531028 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed 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 false colocalization colocalization false false false fig.4a When UBIAD1 and H-Ras were overexpressed in HEK293T cells, both molecules colocalized in the Golgi apparatus (Fig. 4a). hras-ubiad1-7 3531024 N/A 3531028 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell bfp tag fluorescent prot tag fluorescent protein tag fluorescent prot tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3531027 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed 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 false colocalization colocalization false false false suppl. fig.6c statins were also able to disrupt the function of Ras because FPP is necessary for the post-translational modification of Ras (Supplementary Fig. S6c). Therefore, statins can block the function of both UBIAD1 and Ras, consistent with those of previous studies statins hras-ubiad1-3 3531025 N/A 3531027 unspecified role unspecified role fluorescence donor fluorescence donor over-expressed over expressed level over-expressed living cell living cell bfp tag fluorescent prot tag fluorescent protein tag fluorescent prot tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3531028 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed 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 false proximity proximity close-contact colocalization neighbourhood interaction false false false fig.4b, suppl. fig.3b To provide evidence that UBIAD1 forms a complex with H-Ras, a fluorescence resonance energy transfer (FRET) assay was employed to detect this molecular interaction. Significant FRET signals were detected with 405-515 microscope channels when HEK293T cells were cotransfected with UBIAD1 and H-Ras (Fig. 4b). hras-ubiad1-8 3531026 N/A 3531027 unspecified role unspecified role 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 n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 3531028 unspecified role unspecified role 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 c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false suppl. fig.3c