The Saccharomyces cerevisiae checkpoint gene BUB1 encodes a novel protein kinase.
The Saccharomyces cerevisiae checkpoint gene BUB1 encodes a novel protein kinase.
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酿酒酵母检查点基因 BUB1 编码一种新型蛋白激酶。
DOI：
10.1128/mcb.14.12.8282-8291.1994
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发表时间：
1994
影响因子：
5.3
通讯作者：
Hoyt,MA
中科院分区：
文献类型：
--
作者：
Roberts,BT;Farr,KA;Hoyt,MA
正常的细胞增殖要求有丝分裂事件以仔细有序的方式发生。细胞利用检查点来阻止周期进展，直到某些先决步骤完成。为了探索检查点执行的机制，我们之前筛选了酿酒酵母突变体，这些突变体无法从苯并咪唑相关微管抑制剂的短暂治疗中恢复，因为它们无法抑制后续的细胞周期步骤。两个已鉴定的基因，BUB2 和BUB3，已被克隆和描述（M.A.Hoyt、L.Totis 和 B.T.Roberts，Cell 66：507-517，1991）。在这里，我们介绍了 BUB1 基因及其产物的特征。获得的遗传证据表明 Bub1 和 Bub3 的功能相互依赖，免疫沉淀实验证明了两者之间的物理关联。 BUB1 的序列分析揭示了与蛋白激酶相似的结构域。体外实验证实Bub1具有激酶活性； Bub1 能够自磷酸化并催化 Bub3 的磷酸化。此外，过量产生的 Bub1 被发现定位于细胞核。
Normal cell multiplication requires that the events of mitosis occur in a carefully ordered fashion. Cells employ checkpoints to prevent cycle progression until some prerequisite step has been completed. To explore the mechanisms of checkpoint enforcement, we previously screened for mutants ofSaccharomyces cerevisiaewhich are unable to recover from a transient treatment with a benzimidazole-related microtubule inhibitor because they fail to inhibit subsequent cell cycle steps. Two of the identified genes,BUB2andBUB3,have been cloned and described (M. A. Hoyt, L. Totis, and B. T. Roberts, Cell 66:507-517, 1991). Here we present the characterization of theBUB1gene and its product. Genetic evidence was obtained suggesting that Bub1 and Bub3 are mutually dependent for function, and immunoprecipitation experiments demonstrated a physical association between the two. Sequence analysis ofBUB1revealed a domain with similarity to protein kinases. In vitro experiments confirmed that Bub1 possesses kinase activity; Bub1 was able to autophosphorylate and to catalyze phosphorylation of Bub3. In addition, overproduced Bub1 was found to localize to the cell nucleus.