{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T04:58:07Z","timestamp":1791349087797,"version":"4.3.1"},"reference-count":33,"publisher":"National Academy of Sciences","issue":"23","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[1996,11,12]]},"abstract":"<jats:p>\n                    Small ligand\u2013receptor interactions underlie many fundamental\n processes in biology and form the basis for pharmacological\n intervention of human diseases in medicine. We report herein a genetic\n system, named the yeast three-hybrid system, for detecting\n ligand\u2013receptor interactions\n                    <jats:italic>in vivo<\/jats:italic>\n                    . This system is\n adapted from the yeast two-hybrid system with which a third synthetic\n hybrid ligand is combined. The feasibility of this system was\n demonstrated using as the hybrid ligand a heterodimer of covalently\n linked dexamethasone and FK506. Yeast expressing fusion proteins of the\n hormone binding domain of the rat glucocorticoid receptor fused to the\n LexA DNA-binding domain and of FKBP12 fused to a transcriptional\n activation domain activated reporter genes when plated on medium\n containing the dexamethasone\u2013FK506 heterodimer. The reporter gene\n activation is completely abrogated in a competitive manner by the\n presence of excess FK506. Using this system, we screened a Jurkat cDNA\n library fused to the transcriptional activation domain in yeast\n expressing the hormone binding domain of rat glucocorticoid\n receptor\u2013LexA DNA binding domain fusion protein in the presence of\n dexamethasone\u2013FK506 heterodimer. We isolated overlapping clones of\n human FKBP12. These results demonstrate that the three-hybrid system\n can be used to discover receptors for small ligands and to screen for\n new ligands to known receptors.\n                  <\/jats:p>","DOI":"10.1073\/pnas.93.23.12817","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T10:34:16Z","timestamp":1027679656000},"page":"12817-12821","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":267,"title":["A three-hybrid system for detecting small ligand\u2013protein receptor\u2009interactions"],"prefix":"10.1073","volume":"93","author":[{"given":"Edward\u2009J.","family":"Licitra","sequence":"first","affiliation":[{"name":"Center for Cancer Research, and Departments of Biology\r and Chemistry, Massachusetts Institute of Technology,\r Cambridge, MA 02139"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun\u2009O.","family":"Liu","sequence":"additional","affiliation":[{"name":"Center for Cancer Research, and Departments of Biology\r and Chemistry, Massachusetts Institute of Technology,\r Cambridge, MA 02139"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[1996,11,12]]},"reference":[{"key":"e_1_3_2_1_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.ge.19.120185.001233"},{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.3283939"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1702904"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.116.5.1071"},{"key":"e_1_3_2_5_2","first-page":"1","volume":"46","author":"Jakoby W B","year":"1974","unstructured":"W B Jakoby, M Wilchek Methods Enzymol 46, 1\u2013196 (1974).","journal-title":"Methods Enzymol"},{"key":"e_1_3_2_6_2","volume-title":"The Pharmacological Basis of Therapeutics","author":"Hardman J G","year":"1996","unstructured":"J G Hardman, L E Limbird, P B Molinoff, R W Ruddon, A G 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