Zhu Jin, Heinecke Dominic, Mulla Wahid A, Bradford William D, Rubinstein Boris, Box Andrew, Haug Jeffrey S, Li Rong
Stowers Institute for Medical Research, 1000 East 50th Street, Kansas City, Missouri 64110.
Stowers Institute for Medical Research, 1000 East 50th Street, Kansas City, Missouri 64110 Department of Molecular and Integrative Physiology, University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, Kansas 66160.
G3 (Bethesda). 2015 Mar 30;5(6):1043-56. doi: 10.1534/g3.115.017913.
Errors in mitosis are a primary cause of chromosome instability (CIN), generating aneuploid progeny cells. Whereas a variety of factors can influence CIN, under most conditions mitotic errors are rare events that have been difficult to measure accurately. Here we report a green fluorescent protein-based quantitative chromosome transmission fidelity (qCTF) assay in budding yeast that allows sensitive and quantitative detection of CIN and can be easily adapted to high-throughput analysis. Using the qCTF assay, we performed genome-wide quantitative profiling of genes that affect CIN in a dosage-dependent manner and identified genes that elevate CIN when either increased (icCIN) or decreased in copy number (dcCIN). Unexpectedly, qCTF screening also revealed genes whose change in copy number quantitatively suppress CIN, suggesting that the basal error rate of the wild-type genome is not minimized, but rather, may have evolved toward an optimal level that balances both stability and low-level karyotype variation for evolutionary adaptation.
有丝分裂错误是染色体不稳定（CIN）的主要原因，会产生非整倍体子代细胞。虽然多种因素可影响CIN，但在大多数情况下，有丝分裂错误是罕见事件，难以准确测量。在此，我们报告了一种基于绿色荧光蛋白的出芽酵母定量染色体传递保真度（qCTF）检测方法，该方法能够灵敏且定量地检测CIN，并且可轻松适用于高通量分析。使用qCTF检测方法，我们对以剂量依赖方式影响CIN的基因进行了全基因组定量分析，鉴定出在拷贝数增加（icCIN）或减少（dcCIN）时会升高CIN的基因。出乎意料的是，qCTF筛选还揭示了拷贝数变化能定量抑制CIN的基因，这表明野生型基因组的基础错误率并未降至最低，而是可能已朝着一个最佳水平进化，该水平在稳定性和低水平核型变异之间取得平衡，以实现进化适应。
