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酿酒酵母菌落生长与衰老:伴随基因表达变化的双相生长

Saccharomyces cerevisiae colony growth and ageing: biphasic growth accompanied by changes in gene expression.

作者信息

Meunier J R, Choder M

机构信息

Department of Cell Biology, Weizmann Institute of Science, Rehovot 76100, Israel.

出版信息

Yeast. 1999 Sep 15;15(12):1159-69. doi: 10.1002/(SICI)1097-0061(19990915)15:12<1159::AID-YEA441>3.0.CO;2-D.

Abstract

Although colony growth and morphology are central tools in yeast genetics, little is known about the cell physiology and how it changes during the colony growth and ageing. Here we show that the growth of a well-separated Saccharomyces cerevisiae colony is biphasic; a rapid growth phase is followed by a sharp transition to a slower growth phase. In the first growth phase ( approximately 24 cell divisions) most, if not all, cells divide at a rate similar to that in liquid medium and exhibit morphological, biochemical and genetic characteristics of cells engaged in the cell cycle. During the second growth phase, cells in the centre of a colony gradually enter stationary phase, so that later in this phase the growth occurs predominantly at the periphery. Unlike the biphasic growth in rich liquid media containing a fermentable carbon source, in which the first growth is fueled by fermentation and the second by aerobic metabolism, the two phases of the colony growth can be fueled either exclusively by fermentation or exclusively by aerobic metabolism. We also describe a novel technique for in situ estimation of the transcriptional status in the colony cells, which was used to monitor transcription dynamics during the colony development. Using this technique and standard methods to determine mRNA levels, we show that the transition between the first and second growth phases is accompanied by a global change in the pattern of transcription: transcription of most genes is repressed while that of some genes is induced.

摘要

尽管菌落生长和形态是酵母遗传学中的核心工具,但对于细胞生理学以及它在菌落生长和老化过程中如何变化却知之甚少。在这里我们表明,一个分离良好的酿酒酵母菌落的生长是双相的;快速生长阶段之后会急剧转变为较慢的生长阶段。在第一个生长阶段(大约24次细胞分裂),大多数(如果不是全部)细胞以与液体培养基中相似的速率分裂,并表现出参与细胞周期的细胞的形态、生化和遗传特征。在第二个生长阶段,菌落中心的细胞逐渐进入静止期,因此在这个阶段后期,生长主要发生在周边。与含有可发酵碳源的丰富液体培养基中的双相生长不同,在这种培养基中,第一次生长由发酵提供能量,第二次由有氧代谢提供能量,菌落生长的两个阶段可以完全由发酵或完全由有氧代谢提供能量。我们还描述了一种用于原位估计菌落细胞转录状态的新技术,该技术用于监测菌落发育过程中的转录动态。使用这种技术和标准方法来确定mRNA水平,我们表明第一和第二生长阶段之间的转变伴随着转录模式的全局变化:大多数基因被抑制,而一些基因被诱导。

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