DIP Database of Interacting Proteins 611 Young Drive East; Los Angeles CA 90095; USA http://dip.doe-mbi.ucla.edu/ dip@mbi.ucla.edu Severe acute respiratory syndrome-associated coronavirus 3a protein forms an ion channel and modulates virus release. Severe acute respiratory syndrome-associated coronavirus 3a protein forms an ion channel and modulates virus release. Proceedings of the National Academy of Sciences of the United States of America 2006 imex curation Lu W., Zheng BJ., Xu K., Schwarz W., Du L., Wong CK., Chen J., Duan S., Deubel V., Sun B. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus bsun@sibs.ac.cn Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Lu W., Zheng BJ., Xu K., Schwarz W., Du L., Wong CK., Chen J., Duan S., Deubel V., Sun B. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus bsun@sibs.ac.cn Proceedings of the National Academy of Sciences of the United States of America 2006 Only protein-protein interactions imex curation Severe acute respiratory syndrome-associated coronavirus 3a protein forms an ion channel and modulates virus release. Severe acute respiratory syndrome-associated coronavirus 3a protein forms an ion channel and modulates virus release. Proceedings of the National Academy of Sciences of the United States of America 2006 imex curation Lu W., Zheng BJ., Xu K., Schwarz W., Du L., Wong CK., Chen J., Duan S., Deubel V., Sun B. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus bsun@sibs.ac.cn Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Lu W., Zheng BJ., Xu K., Schwarz W., Du L., Wong CK., Chen J., Duan S., Deubel V., Sun B. Virus - Publications including interactions involving viral proteins Coronavirus - Interactions investigated in the context of Coronavirus DIP original experiment: DIP-263490X bsun@sibs.ac.cn Proceedings of the National Academy of Sciences of the United States of America 2006 IMEx Severe acute respiratory syndrome-associated coronavirus 3a protein forms an ion channel and modulates virus release. ap3a_sars ORF3a protein Protein 3a Accessory protein 3a Protein U274 Protein X1 3a protein protein sars-cov Human SARS coronavirus SARS-CoV Severe acute respiratory syndrome coronavirus MDLFMRFFTLGSITAQPVKIDNASPASTVHATATIPLQASLPFGWLVIGVAFLAVFQSATKIIALNKRWQLALYKGFQFICNLLLLFVTIYSHLLLVAAGMEAQFLYLYALIYFLQCINACRIIMRCWLCWKCKSKNPLLYDANYFVCWHTHNYDYCIPYNSVTDTIVVTEGDGISTPKLKEDYQIGGYSEDRHSGVKDYVVVHGYFTEVYYQLESTQITTDTGIENATFFIFNKLVKDPPNVQIHTIDGSSGVANPAMDPIYDEPTTTTSVPL DIP protein P59632 original sequence version: 70 ap3a_cvhsa-1 2 3 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept over-expressed over expressed level over-expressed YFP-tag YFP-tag yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range P59632:p.Cys133Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain C A human Homo sapiens Human 3 unspecified role unspecified role fluorescence donor fluorescence donor over-expressed over expressed level over-expressed CFP-tag CFP-tag cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range P59632:p.Cys133Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain C A human Homo sapiens Human proximity proximity close-contact colocalization neighbourhood interaction Figure 3D ap3a_cvhsa-6 1 3 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed C-terminus YFP-tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range P59632:p.Cys81Ala 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 C A P59632:p.Cys117Ala 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 C A P59632:p.Cys121Ala 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 C A P59632:p.Cys127Ala 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 C A P59632:p.Cys130Ala 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 C A P59632:p.Cys133Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain C A P59632:p.Cys148Ala 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 C A P59632:p.Cys157Ala 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 C A 3 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed C-terminus YFP-tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range P59632:p.Cys81Ala 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 C A P59632:p.Cys117Ala 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 C A P59632:p.Cys121Ala 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 C A P59632:p.Cys127Ala 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 C A P59632:p.Cys130Ala 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 C A P59632:p.Cys133Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain C A P59632:p.Cys148Ala 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 C A P59632:p.Cys157Ala 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 C A association association Figure 3 A/C After treatment with DTT, the bands of 75 kDa and 150 kDa disappeared. DTT ap3a_cvhsa-ap3a_cvhsa-2 1 3 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed C-terminus YFP-tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range P59632:p.Cys81Ala 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 C A P59632:p.Cys117Ala 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 C A P59632:p.Cys121Ala 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 C A P59632:p.Cys127Ala 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 C A P59632:p.Cys130Ala 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 C A P59632:p.Cys133Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain C A P59632:p.Cys148Ala 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 C A P59632:p.Cys157Ala 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 C A 3 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed C-terminus YFP-tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range P59632:p.Cys81Ala 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 C A P59632:p.Cys117Ala 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 C A P59632:p.Cys121Ala 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 C A P59632:p.Cys127Ala 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 C A P59632:p.Cys130Ala 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 C A P59632:p.Cys133Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain C A P59632:p.Cys148Ala 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 C A P59632:p.Cys157Ala 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 C A association association Figure 3 A/C After treatment with DTT, the bands of 75 kDa and 150 kDa disappeared. DTT