합성 스캐폴드 단백질을 이용한 MAPK 신호전달의 재구성과 인위적 조작

Abstract

Acting as a hub for eukaryotic cell signaling, scaffold proteins are attractive targets for engineering and manipulating signaling circuits. The ultimate experimental challenge to this premise is to reconstruct a signaling pathway using a rationally designed scaffold with modular architecture. Here, we designed synthetic scaffolds to reorganize the MAPK pathway involved in mating in yeast. The synthetic scaffolds composed of modular interaction domains recruited three member kinases to the plasma membrane and were capable of generating the signaling responses by functioning as logic gates of signaling circuits. The modular architecture of the synthetic scaffold assembly allowed the artificial manipulation of signaling behaviors, including the modulation of signaling strength and the incorporation of a negative control. Furthermore, this study demonstrated that three tiers of the MAPK pathway exhibited decreasing positional plasticity from the top (MAPKKK) to the bottom (MAPK) tier, suggesting that novel pathway connectivities could be generated. This synthetic approach with designer scaffolds should allow the rational manipulation or engineering of signaling pathways and provide insight into the functional mechanisms of scaffold proteins and scaffold-dependent pathway evolution.