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 Overexpression of inactive tetherin delGPI mutant inhibits HIV-1 Vpu-mediated antagonism of endogenous tetherin. Overexpression of inactive tetherin delGPI mutant inhibits HIV-1 Vpu-mediated antagonism of endogenous tetherin. FEBS Lett. (0014-5793) 2013 Lv M., Wang J., Zhu Y., Wang X., Zuo T., Liu D., Zhang J., Wu J., Zhang H., Kong W., Yu X. 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 Lv M., Wang J., Zhu Y., Wang X., Zuo T., Liu D., Zhang J., Wu J., Zhang H., Kong W., Yu X. FEBS Lett. (0014-5793) 2013 Overexpression of inactive tetherin delGPI mutant inhibits HIV-1 Vpu-mediated antagonism of endogenous tetherin. Overexpression of inactive tetherin delGPI mutant inhibits HIV-1 Vpu-mediated antagonism of endogenous tetherin. FEBS Lett. (0014-5793) 2013 Lv M., Wang J., Zhu Y., Wang X., Zuo T., Liu D., Zhang J., Wu J., Zhang H., Kong W., Yu X. 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 anti tag immunost anti tag immunostaining anti tag immunost Lv M., Wang J., Zhu Y., Wang X., Zuo T., Liu D., Zhang J., Wu J., Zhang H., Kong W., Yu X. FEBS Lett. (0014-5793) 2013 bst2_human Bone marrow stromal antigen 2 BST2 Tetherin HM1.24 antigen protein protein human Homo sapiens Human MASTSYDYCRVPMEDGDKRCKLLLGIGILVLLIIVILGVPLIIFTIKANSEACRDGLRAVMECRNVTHLLQQELTEAQKGFQDVEAQAATCNHTVMALMASLDAEKAQGQKKVEELEGEITTLNHKLQDASAEVERLRRENQVLSVRIADKKYYPSSQDSSSAAAPQLLIVLLGLSALLQ DIP protein Q10589 original sequence version: 87 DIP protein Q10589 original sequence version: 93 DIP protein Q10589 original sequence version: 92 CAF52340D69061EE vpu_hv1br Protein Vpu vpu U ORF protein Viral protein U protein protein hv1br Human immunodeficiency virus type 1 group M subtype B (isolate BRU/LAI) HIV-1 MQPIQIAIAALVVAIIIAIVVWSIVIIEYRKILRQRKIDRLIDRLIERAEDSGNESEGEISALVEMGVEMGHHAPWDIDDL mint homomint B9F588F7C6654B51 bst2-vpu 1 Vpu 4 anti tag western anti tag western blot anti tag western experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Tetherin 3 anti tag western anti tag western blot anti tag western experimental particp experimental participant identification experimental particp 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f2b homomint mint Next, the interaction of delGPI tetherin with Vpu was verified by immunoprecipitation. 293T cells were co-transfected with the tetherin delGPI or tetherin AGMTM delGPI plasmid along with the Vpu-cmyc plasmid. Cells were lysed and incubated with HA-beads to precipitate HA-tagged tetherin. As shown in Fig. 2B, expression levels of Vpu in the three samples were comparable, and those of delGPI and AGMTM delGPI tetherin were similar as well (pre-immunoprecipitation lysates). Vpu was nearly absent in the mock control immunoprecipitation sample. Meanwhile, Vpu was pulled down with tetherin delGPI. However, when tetherin AGMTM delGPI was precipitated, the Vpu band was much weaker than that with tetherin delGPI. This result confirmed that the tetherin delGPI mutant maintained Vpu binding activity. The tetherin mutant lacking the GPI anchor (delGPI) was generated by replacing codon 161 with a termination codon. The human tetherin gene with the TM sequence replaced by the synthetic AGMTM sequence bst2-vpu-1 2 Vpu 4 anti tag immunost anti tag immunostaining anti tag immunost experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Tetherin 3 anti tag immunost anti tag immunostaining anti tag immunost experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization f2a homomint mint As shown in Fig. 2A, the distribution of tetherin delGPI presented as dense punctas with a tendency to spread outward from the perinuclear area. This pattern indicates that the protein mainly accumulated within the recycling endosome, which is largely consistent with wild-type tetherin [14]. Furthermore, the distribution patterns of tetherin delGPI and Vpu partially overlapped, indicating a certain degree of co-localization. The tetherin AGMTM delGPI showed quite a similar pattern, indicating that replacement of its TM domain may result in loss of sensitivity to 1 Vpu but not from the gross change in subcellular localization.