Feiler H S, Jacobs T W
Department of Plant Biology, University of Illinois, Urbana 61801.
Proc Natl Acad Sci U S A. 1990 Jul;87(14):5397-401. doi: 10.1073/pnas.87.14.5397.
The mitotic cell cycle of yeast and animal cells is regulated by the cdc2 gene and its product, the p34 protein kinase, and by other components of the MPF or histone H1 kinase complex. We present evidence that cdc2, p34, and a histone H1 kinase also exist in higher plants. Protein extracts from 10 plant species surveyed display a 34-kDa component recognized by a monoclonal antibody directed against an evolutionarily conserved epitope of fission yeast p34. Nondenatured protein extracts of mitotic Pisum sativum (garden pea) tissues were fractionated by gel filtration, electrophoretically separated under denaturing conditions, and immunoblotted. p34 crossreactive material was apparent in both low and high molecular mass fractions, indicating that pea p34 occurs as both a monomer and as part of a high molecular mass complex. Histone H1 kinase activity was found predominantly in the higher molecular mass fractions, those to which the least phosphorylated form of pea p34 was confined. We also report the cloning of the pea homologue of cdc2 by polymerase chain reaction. DNA sequence analysis reveals perfect conservation of the hallmark "PSTAIR" sequence motif found in all cdc2 gene products analyzed to date.
酵母和动物细胞的有丝分裂细胞周期受cdc2基因及其产物p34蛋白激酶以及MPF或组蛋白H1激酶复合物的其他成分调控。我们提供的证据表明,cdc2、p34和组蛋白H1激酶也存在于高等植物中。对10种被调查植物物种的蛋白质提取物进行检测,结果显示有一种34 kDa的成分,可被一种针对裂殖酵母p34进化保守表位的单克隆抗体识别。对有丝分裂期豌豆组织的非变性蛋白质提取物进行凝胶过滤分级分离,在变性条件下进行电泳分离并进行免疫印迹分析。在低分子量和高分子量组分中均可见p34交叉反应物质,这表明豌豆p34既以单体形式存在,也作为高分子量复合物的一部分存在。组蛋白H1激酶活性主要存在于高分子量组分中,即豌豆p34磷酸化程度最低的形式所在的组分。我们还报告了通过聚合酶链反应克隆豌豆cdc2同源物的情况。DNA序列分析显示,在迄今为止分析的所有cdc2基因产物中发现的标志性“PSTAIR”序列基序完全保守。