Abstract
Progression through the cell division cycle is orchestrated by a complex network of interacting genes and proteins. Some of these proteins are known to fluctuate periodically during the cell cycle, but a systematic study of the fluctuations of a broad sample of cell-cycle proteins has not been made until now. Using time-lapse fluorescence microscopy, we profiled 16 strains of budding yeast, each containing GFP fused to a single gene involved in cell cycle regulation. The dynamics of protein abundance and localization were characterized by extracting the amplitude, period, and other indicators from a series of images. Oscillations of protein abundance could clearly be identified for Cdc15, Clb2, Cln1, Cln2, Mcm1, Net1, Sic1, and Whi5. The period of oscillation of the fluorescently tagged proteins is generally in good agreement with the inter-bud time. The very strong oscillations of Net1 and Mcm1 expression are remarkable since little is known about the temporal expression of these genes. By collecting data from large samples of single cells, we quantified some aspects of cell-to-cell variability due presumably to intrinsic and extrinsic noise affecting the cell cycle.
Highlights
The cell division cycle is the sequence of events whereby a living cell replicates its components and divides them between two daughter cells, so that each daughter receives the information and machinery necessary to repeat the process
The unfiltered and filtered time-course of a typical cell for each of the 16 yeast strains are shown in Figure 2, along with a wild-type (WT) control cell not labeled with GFP
For instance the fluorescence signals collected on the Cln2 and Clb2 tagged strains are well correlated with the cell budding event
Summary
The cell division cycle is the sequence of events whereby a living cell replicates its components and divides them between two daughter cells, so that each daughter receives the information and machinery necessary to repeat the process. The sequence of events is very tightly controlled, the time taken to progress through each stage of the cell cycle may vary considerably from cell to cell. Previous research into the expression of genes controlling progression through the eukaryotic cell cycle has heavily relied on bulk measurements, such as western (and northern) blots and micro-arrays, on populations of cells that have been synchronized by some strong perturbation, for examples see the experimental data used in the development of the model of Chen et al [5]
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