PKA-regulated phosphorylation status of S149 and S321 sites of CDC25B inhibits mitosis of fertilized mouse eggs
- Author:
- No author available
- Journal Title:
- Acta Physiologica Sinica
- Issue:
- 1
- DOI:
- No doi available
- Key Word:
- 细胞分裂周期25B;蛋白激酶A;有丝分裂成熟促进因子;H-89;小鼠受精卵
Abstract： 在小鼠1-细胞期受精卵,蛋白激酶A (protein kinase A,PKA)可通过磷酸化细胞分裂周期25B (cell division cycle 25B,CDC25B)的321位Ser引起受精卵分裂阻滞.本文旨在研究PKA对CDC25B 149位点Ser的磷酸化对小鼠受精卵发育的影响,及其与321位点的关系.质粒pBSK-CDC25B-WT (野生型)、pBSK-CDC25B-S149A (149位Ser突变成Ala)、pBSK-CDC25B-S321A (321位Ser突变成Ala)、pBSK-CDC25B-S 149A/S321A (149位和321位Ser联合突变成Ala)体外转录成mRNAs;显微注射入小鼠S期受精卵中,在PKA抑制剂H-89预处理的M16培养基中培养,观察其对受精卵发育、有丝分裂成熟促进因子(maturation promoting factor,MPF)活性及CDC2-Tyrl5磷酸化状态的影响.结果显示,H-89呈剂量(0～50 μmol/L)依赖性的方式加速小鼠受精卵卵裂.然后选择卵裂率接近100％、H-89 (40 μmol/L)培养的小鼠受精卵,与对照组相比,CDC25B各种mRNAs注射组受精卵卵裂率明显增高,MPF活性提前达到高峰；CDC2-Tyrl5磷酸化状态和MPF活性变化相一致,以上结果说明,CDC25B蛋白的149位Ser也是PKA在体内调控CDC25B的重要靶点.因此在小鼠1-细胞期受精卵有丝分裂过程中,PKA对小鼠CDC25B蛋白S149位点与S321位点的磷酸化修饰可能是控制受精卵G2/M转换的重要方式.
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