Interdisciplinary Research Unit , National Institute for Basic Biology (NIBB), National Institutes of Natural Sciences (NINS), Okazaki, Aichi, 444-8585, Japan.
Basic Biology Program, SOKENDAI (The Graduate University for Advanced Studies), Hayama, Kanagawa, 240-0193, Japan.
J Cell Sci. 2024 Feb 15;137(4). doi: 10.1242/jcs.261625. Epub 2024 Feb 28.
Certain proteins assemble into diverse complex states, each having a distinct and unique function in the cell. Target of rapamycin (Tor) complex 1 (TORC1) plays a central role in signalling pathways that allow cells to respond to the environment, including nutritional status signalling. TORC1 is widely recognised for its association with various diseases. The budding yeast Saccharomyces cerevisiae has two types of TORC1, Tor1-containing TORC1 and Tor2-containing TORC1, which comprise different constituent proteins but are considered to have the same function. Here, we computationally modelled the relevant complex structures and then, based on the structures, rationally engineered a Tor2 mutant that could form Tor complex 2 (TORC2) but not TORC1, resulting in a redesign of the complex states. Functional analysis of the Tor2 mutant revealed that the two types of TORC1 induce different phenotypes, with changes observed in rapamycin, caffeine and pH dependencies of cell growth, as well as in replicative and chronological lifespan. These findings uncovered by a general approach with huge potential - model structure-based engineering - are expected to provide further insights into various fields such as molecular evolution and lifespan.
某些蛋白质组装成不同的复杂状态,每种状态在细胞中都具有独特的功能。雷帕霉素靶蛋白(TOR)复合物 1(TORC1)在信号通路中发挥着核心作用,使细胞能够对环境做出反应,包括营养状态信号。TORC1 广泛与各种疾病相关联。酿酒酵母有两种类型的 TORC1,即含有 Tor1 的 TORC1 和含有 Tor2 的 TORC1,它们包含不同的组成蛋白,但被认为具有相同的功能。在这里,我们通过计算建模了相关的复合物结构,然后基于这些结构,对 Tor2 突变体进行了合理的工程设计,使其能够形成 TORC2,但不能形成 TORC1,从而重新设计了复合物状态。对 Tor2 突变体的功能分析表明,两种类型的 TORC1 诱导不同的表型,观察到雷帕霉素、咖啡因和 pH 值依赖性细胞生长以及复制和生物钟寿命的变化。这种通过通用方法揭示的发现——基于模型结构的工程设计——有望为分子进化和寿命等各个领域提供进一步的见解。