York J D, Majerus P W
Division of Hematology-Oncology, Washington University School of Medicine, St. Louis, Missouri 63110.
J Biol Chem. 1994 Mar 18;269(11):7847-50.
In the current study we have measured phosphatidylinositols during the cell cycle. HeLa cells were labeled with [3H]myoinositol to a steady state, synchronized to the G1/S boundary, and the levels of phosphatidylinositol (PtdIns) lipids were measured at various times after release from the block. The levels of total cellular PtdIns, PtdIns(4)P, and PtdIns(4,5)P2 relative to total cellular phospholipid did not vary throughout the cell cycle. We then isolated nuclei from synchronized cells using a non-detergent method and found that the levels of nuclear PtdIns lipids decreased by over 50% at 2 and 4 h after release from the G1/S boundary (S-phase of the cell cycle) and returned to the original levels by 9 h. Separation of individual inositol-containing nuclear lipids showed that PtdIns decreased by 50% while levels of PtdIns(4)P and PtdIns(4,5)P2 decreased by 66%. Levels of the cytoplasmic PtdIns lipids remained constant throughout this period. This experiment indicates that there is specific nuclear. PtdIns turnover that is activated during DNA synthesis.
在当前研究中,我们在细胞周期中测量了磷脂酰肌醇。将HeLa细胞用[3H]肌醇标记至稳态,同步至G1/S边界,并在从阻滞中释放后的不同时间测量磷脂酰肌醇(PtdIns)脂质的水平。相对于总细胞磷脂,总细胞PtdIns、PtdIns(4)P和PtdIns(4,5)P2的水平在整个细胞周期中没有变化。然后,我们使用非去污剂方法从同步化细胞中分离细胞核,发现从G1/S边界(细胞周期的S期)释放后2小时和4小时,核PtdIns脂质水平下降超过50%,并在9小时恢复到原始水平。对单个含肌醇的核脂质进行分离显示,PtdIns下降了50%,而PtdIns(4)P和PtdIns(4,5)P2的水平下降了66%。在此期间,细胞质PtdIns脂质水平保持恒定。该实验表明,在DNA合成过程中存在特定的核PtdIns周转被激活。