Defining Emergence from Stationary Phase in Yeast

描述(申请人提供)：本研究的目的是了解酿酒酵母中静止细胞以及静止细胞和有丝分裂周期细胞之间的关系。在基因组学出现之前，在转录组水平上研究这些问题是不可能的。使用最近开发的快速采样技术，我们可以第一次对处于静止状态的细胞培养物和指数分裂的细胞培养物中的基因表达进行精细分辨率的时间过程研究。为了检查静止细胞及其与循环细胞的关系，我们将：1.)研究固定相培养在细胞大小、DNA含量和基因表达方面的异质性，2。在同步分裂单元中进行G1的快速采样，3.)从细胞退出静止期开始进行更长的时间进程，以及4.)开发和使用多元曲线解析方法作为强大的基因组分析工具。通过了解酵母中与静止有关的问题，我们可以对折磨人类的疾病有重要的见解，例如由静止的细菌病原体引起的慢性感染，以及在已经失去保持静止的能力的细胞中观察到的致癌转化。

DESCRIPTION (provided by applicant): The objective of the research proposed here is to understand quiescent cells, and the relationship between quiescent cells and mitotically cycling cells in the budding yeast Saccharomyces cerevisiae. Prior to the advent of genomics, it was not possible to study these questions at the level of the transcriptome. Using recently developed rapid-sampling technology; we can generate, for the first time, fine-resolution time course studies of gene expression in cell cultures exiting quiescence, and in exponentially dividing cell cultures. To examine quiescent cells and the relationship to cycling cells we will: 1.) Investigate the heterogeneity of stationary phase cultures with respect to cell size, DNA content, and gene expression, 2.) Conduct a rapid sampling of G1 in synchronized dividing cells, 3.) Conduct a longer time course from cells exiting stationary phase, and 4.) Develop and use mutivariate curve resolution methods as a powerful genomic analysis tool. By understanding the questions related to quiescence in yeast, we can gain significant insights into diseases that afflict humans such as the chronic infections caused by quiescent bacterial pathogens, and the carcinogenic transformation observed in cells that have lost the capacity to remain quiescent.