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Journal articles
A Conditionally Expressed Third Partner Stabilizes or Prevents the Formation of a Transcriptional Activator in a Three-hybrid System
Abstract : We describe a three-hybrid system that involves three polypeptides that allow or prevent the formation of the transcriptional activator. Beside the two-hybrid fusion proteins, the third partner is under the control of the Met25 promoter, which is positively regulated in medium lacking methionine. We document a situation where such a third partner promotes interaction between two proteins, one fused to a DNA-binding domain and the other fused to an activator domain. This is demonstrated for cdk7-MAT1 interaction stabilized by the presence of cyclin H; these three polypeptides are found either free or associated with the transcription/DNA repair factor TFIIH. We also document the capacity of our system to conditionally inhibit the interaction between two polypeptides that otherwise elicit a positive two-hybrid response. This is demonstrated for Ras-Raf interaction precluded by an excess of Raf. The presence of a methionine-regulated promoter provides an "on" or "off" switch for the formation of the transcriptional activator, thus also providing an excellent control to evaluate the activation or inhibition properties of the third partner.
Cited literature [29 references]
https://www.hal.inserm.fr/inserm-02438566
Contributor : Franck Tirode <>
Submitted on : Tuesday, January 14, 2020 - 11:51:31 AM
Last modification on : Thursday, June 17, 2021 - 3:16:03 PM
Long-term archiving on: : Wednesday, April 15, 2020 - 5:09:31 PM
Contributor : Franck Tirode <>
Submitted on : Tuesday, January 14, 2020 - 11:51:31 AM
Last modification on : Thursday, June 17, 2021 - 3:16:03 PM
Long-term archiving on: : Wednesday, April 15, 2020 - 5:09:31 PM