Supplementary MaterialsS1 Fig: Characterization of temperature- or optogenetically-induced isotropic growth

Supplementary MaterialsS1 Fig: Characterization of temperature- or optogenetically-induced isotropic growth. G1 (i.e., Fig E, F, and G in S1 Fig). (I) Fluorescence of exogenously-expressed PhyB-mCherry-Tom7 under control of an ADH1 promoter was measured in cells of indicated volumes. Cells were binned by mother volume in 200-m increments. The average volume within each bin is usually plotted. N = 300 cells. Error bars, SD. r, Pearsons correlation coefficient. (J) Growth rates of single cells at 37C. Cells were shifted from 25C to 37C 45 min prior to the start of the experiment to allow for Cdk1 disruption.(TIF) pone.0209301.s001.TIF (1020K) GUID:?EB195097-97B7-4567-AFB9-29AFE459B420 S2 Fig: Volume SPDB-DM4 measurements of daughter cells. (A) Representative optoBem1 daughter cells from experiments in Fig 4C and 4D. Only the daughters of daughters were measured for each generation. (B) Histograms depicting cell volume distributions for indicated timepoints in Fig 3A.(TIF) pone.0209301.s002.TIF (169K) GUID:?0C3B2D77-710A-4F20-8D9C-A58618EA2F87 S3 Fig: Growth measurements of SPDB-DM4 yeast strains. (A) opto-Bem1 cells were illuminated for 6C8 h with red light (to generate giant yeast), then switched to IR light (allowing giant yeast to bud and divide). Similarly, cells were incubated at 37C for 8 h (to generate giant yeast), then shifted to 25C (allowing giant yeast to bud and divide). All cells were imaged every SPDB-DM4 5C10 min for ~8 h. Exogenously-expressed Cdc10-GFP was used to mark septin rings (green) and measure cell cycle progression. Panels depict representative opto-Bem1 cells. Budding duration, difference between the time of division (e.g., septin ring disappearance at 01:45) and time of birth (e.g., septin ring appearance at 00:30). Mother volume was measured at the time of daughter cell birth (e.g., yellow arrow) and daughter volume (i.e. only the former bud compartment) was measured at cytokinesis (e.g., blue arrow). Time, HH:MM. (B) Doubling occasions of indicated strains in liquid culture at 25C during log-phase growth.(TIF) pone.0209301.s003.TIF (456K) GUID:?DED4C531-21EA-4963-BD06-FCDD1CDD003E S1 Supporting Information: (PDF) pone.0209301.s004.pdf (78K) GUID:?DB4E3719-4E2D-4A76-A3BA-45FC65625A31 Data Availability StatementAll relevant data are within the manuscript Rabbit Polyclonal to SFRS17A and its Supporting Information file. Abstract Cell populations across nearly all forms of life generally maintain a characteristic cell type-dependent size, but how size control is usually achieved has been a long-standing question. The G1/S boundary of the cell cycle serves as a major point of size control, and mechanisms operating here restrict SPDB-DM4 passage of cells to Start if they are too small. In contrast, it is less clear how size is usually regulated post-Start, during S/G2/M. To gain further insight into post-Start size control, we prepared budding yeast that can be reversibly blocked from bud initiation. While blocked, cells continue to grow isotropically, increasing their volume by more than an order of magnitude over unperturbed cells. Upon release from their block, giant mothers reenter the cell cycle and their progeny rapidly return to the original unperturbed size. We found this behavior to be consistent with a size-invariant timer specifying the duration of S/G2/M. These results indicate that yeast use at least two distinct mechanisms at different cell cycle phases to ensure size homeostasis. Introduction Cell size correlates strongly with key aspects of cell physiology, including organelle abundance [1,2] and DNA ploidy [3]. Maintenance of uniform size may also underlie the efficient functioning of tissues and organs [4]. While cells employ diverse strategies to.