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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >G1 cyclin-dependent activation of p34CDC28 (Cdc28p) in vitro.
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G1 cyclin-dependent activation of p34CDC28 (Cdc28p) in vitro.

机译：G1细胞周期蛋白依赖性激活p34CDC28（Cdc28p）在体外。

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摘要

In Saccharomyces cerevisiae, transient accumulation of G1 cyclin/p34CDC28 (Cdc28p) complexes induces cells to traverse the cell cycle Start checkpoint and commit to a round of cell division. To investigate posttranslational controls that modulate Cdc28p activity during the G1 phase, we have reconstituted cyclin-dependent activation of Cdc28p in a cyclin-depleted G1 extract. A glutathione S-transferase-G1 cyclin chimera (GST-Cln2p) efficiently binds to and activates Cdc28p as a histone H1 kinase. Activation of Cdc28p by GST-Cln2p requires ATP, crude yeast cytosol, and the conserved Thr-169 residue that serves in other organisms as a substrate for phosphorylation by cyclin-dependent protein kinase-activating kinase. This assay may be useful for distinguishing genes that promote directly the posttranslational assembly of active Cln2p/Cdc28p kinase complexes from those that stimulate the accumulation of active complexes via a positive-feedback loop that governs synthesis of G1 cyclins.
机译：在酿酒酵母中，G1细胞周期蛋白/ p34CDC28（Cdc28p）复合物的瞬时积累会诱导细胞穿越细胞周期起始检查点并进行一轮细胞分裂。为了研究在G1阶段调节Cdc28p活性的翻译后对照，我们重建了细胞周期蛋白缺失的G1提取物中Cdc28p的细胞周期蛋白依赖性激活。谷胱甘肽S-转移酶-G1细胞周期蛋白嵌合体（GST-Cln2p）有效结合并激活Cdc28p作为组蛋白H1激酶。 GST-Cln2p激活Cdc28p需要ATP，粗酵母胞质溶胶和保守的Thr-169残基，该残基在其他生物体中充当细胞周期蛋白依赖性蛋白激酶激活激酶磷酸化的底物。该测定法可能有助于区分直接促进活性Cln2p / Cdc28p激酶复合物的翻译后组装的基因与那些通过控制G1细胞周期蛋白合成的正反馈环刺激活性复合物积累的基因。

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