Drug Discovery Research Center (DDRC), Translational Health Science and Technology Institute (THSTI), NCR Biotech Science Cluster, Faridabad, 121001, India.
Department of Zoology, University of Delhi, Delhi, 110007, India.
Sci Rep. 2018 Jan 22;8(1):1303. doi: 10.1038/s41598-018-19689-0.
Cell growth and proliferation are two diverse processes yet always linked. Akt1, a serine/threonine kinase, is a multi-functional protein implicated in regulation of cell growth, survival and proliferation. Though it has a role in G1/S progression, the manner by which Akt1 controls cell cycle and blends cell growth with proliferation is not well explored. In this study, we characterize the Akt1 interactome as the cell cycle progresses from G0 to G1/S and G2 phase. For this, Akt1-overexpressing HEK293 cells were subjected to AP-MS. To distinguish between individual cell cycle stages, cells were cultured in the light, medium and heavy labelled SILAC media. We obtained 213 interacting partners of Akt1 from these studies. GO classification revealed that a significant number of proteins fall into functional classes related to cell growth or cell cycle processes. Of these, 32 proteins showed varying association with Akt1 in different cell cycle stages. Further analyses uncovered a subset of proteins showing counteracting effects so as to tune stage-specific progression through the cycle. Thus, our study provides some novel perspectives on Akt1-mediated regulation of the cell cycle and offers the framework for a detailed resolution of the downstream cellular mechanisms that are mediated by this kinase.
细胞生长和增殖是两个不同的过程，但总是相关联的。Akt1 是一种丝氨酸/苏氨酸激酶，是一种多功能蛋白，参与调节细胞生长、存活和增殖。尽管它在 G1/S 进展中发挥作用，但 Akt1 控制细胞周期并将细胞生长与增殖融合的方式尚未得到充分探索。在这项研究中，我们在细胞从 G0 到 G1/S 和 G2 期进展的过程中，对 Akt1 相互作用组进行了特征描述。为此，我们用 Akt1 过表达的 HEK293 细胞进行了 AP-MS 实验。为了区分各个细胞周期阶段，我们将细胞在轻、中、重标记的 SILAC 培养基中培养。从这些研究中，我们获得了 213 个 Akt1 的相互作用伙伴。GO 分类表明，大量蛋白质属于与细胞生长或细胞周期过程相关的功能类别。其中，32 种蛋白质在不同的细胞周期阶段与 Akt1 的结合程度不同。进一步的分析揭示了一组具有拮抗作用的蛋白质，以调节通过周期的特定阶段进展。因此，我们的研究为 Akt1 介导的细胞周期调控提供了一些新的视角，并为详细解析该激酶介导的下游细胞机制提供了框架。
