Differential scaling between G1 protein production and cell size dynamics promotes commitment to the cell division cycle in budding yeast

Litsios, Athanasios; Huberts, Daphne H. E. W.; Terpstra, Hanna M; Guerra, Paolo; Schmidt, Alexander; Buczak, Katarzyna; Papagiannakis, Alexandros; Rovetta, Mattia; Hekelaar, Johan; Hubmann, Georg; Exterkate, Marten; Milias-Argeitis, Andreas; Heinemann, Matthias

doi:10.1038/s41556-019-0413-3

Cited by 73 publications

(135 citation statements)

References 81 publications

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“…While our conclusions rely on data from a single cell line, L1210, the growth profiles of L1210s and adherent cells have been shown to be similar (3), suggesting 195 similar cell cycle dependent growth regulation across multiple cell types. Notably, our results do not exclude growth regulation by dilution effects in G1 stage of cell cycle, nor do our results exclude dilution/concentration effects regulating cell cycle progression or cell metabolism(4,(14)(15)(16)(17)(18)40). In fact, size-dependent dilution effects are likely to be responsible for cell size homeostasis, as our data show that cell size homeostasis is not achieved simply by coupling cell growth efficiency to the 200 absolute size of cells.…”

contrasting

confidence: 65%

“…Importantly, such transport limitations can exist even when cellular components scale isometrically with cell size. In a developmental setting, growth-influencing transport limitations could have a major impact on cell physiology, possibly explaining why most fast growing and proliferating cell types are small (<20 40 µm in diameter) (5,6,8). Transport limitations are also considered to result in allometric scaling of metabolism, a phenomenon where larger animals display lower metabolic and growth rates (10,11).…”

Section: Introductionmentioning

confidence: 99%

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Mass measurements of polyploid lymphocytes reveal that growth is not size limited but depends strongly on cell cycle

Kang

Ölçüm

et al. 2019

Preprint

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Cell size is believed to influence cell growth and metabolism. Consistently, several studies have revealed that large cells have lower mass accumulation rates per unit mass (i.e. growth efficiency) than intermediate sized cells in the same population. Size-dependent growth is commonly attributed to transport limitations, such as increased diffusion timescales and decreased surface-to-volume ratio. 5However, separating cell size and cell cycle dependent growth is challenging. To decouple and quantify cell size and cell cycle dependent growth effects we monitor growth efficiency of freely proliferating and cycling polyploid mouse lymphocytes with high resolution. To achieve this, we develop large-channel suspended microchannel resonators that allow us to monitor mass of single cells ranging from 40 pg (small diploid lymphocyte) to over 4000 pg, with a resolution ranging from 10 ~1% to ~0.05%. We find that mass increases exponentially with respect to time in early cell cycle but transitions to linear dependence during late S and G2 stages. This growth behavior repeats with every endomitotic cycle as cells grow in to polyploidy. Overall, growth efficiency changes 29% due to cell cycle. In contrast, growth efficiency did not change due to cell size over a 100-fold increase in cell mass during polyploidization. Consistently, growth efficiency remained constant when cell cycle was 15 arrested in G2. Thus, cell cycle is a primary determinant of growth efficiency and increasing cell size does not impose transport limitations that decrease growth efficiency in cultured mammalian cells. Significance statementCell size is believed to influence cell behavior through limited transport efficiency in larger cells, 20 which could decrease the growth rate of large cells. However, this has not been experimentally investigated due to a lack of non-invasive, high-precision growth quantification methods suitable for measuring large cells. Here, we have engineered large versions of microfluidic mass sensors called suspended microchannel resonators in order to study the growth of single mammalian cells that range 100-fold in mass. This revealed that the absolute size of a cell does not impose strict transport or other 25 limitations that would inhibit growth. In contrast to cell size, however, cell cycle has a relatively large influence on growth and our measurements allow us to decouple and quantify the growth effects caused by cell cycle and cell size.

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contrasting

confidence: 65%

Section: Introductionmentioning

confidence: 99%

Mass measurements of polyploid lymphocytes reveal that growth is not size limited but depends strongly on cell cycle

Kang

Ölçüm

et al. 2019

Preprint

View full text Add to dashboard Cite

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“…Proteomic data also unveiled an increase in protein synthesis activity before cell division, in G1, which could be supported by the increase in amino acids that was observed in the same phase. Litsios et al (28), using microfluidics and time-lapse microscopy in single cells of S. cerevisiae, found that cells similarly demonstrate a pulse in protein production rate during G1. We also find that central carbon metabolism, as shown by an increase in protein abundance, increased after cell division was committed to, most notably with glycolytic pathway enzymes increasing in relative abundance in G2/M.…”

Section: Discussionmentioning

confidence: 99%

Building blocks are synthesized on demand during the yeast cell cycle

Campbell

Westholm

Kasvandik

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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For cells to replicate, a sufficient supply of biosynthetic precursors is needed, necessitating the concerted action of metabolism and protein synthesis during progressive phases of cell division. A global understanding of which biosynthetic processes are involved and how they are temporally regulated during replication is, however, currently lacking. Here, quantitative multiomics analysis is used to generate a holistic view of the eukaryal cell cycle, using the budding yeastSaccharomyces cerevisiae. Protein synthesis and central carbon pathways such as glycolysis and amino acid metabolism are shown to synchronize their respective abundance profiles with division, with pathway-specific changes in metabolite abundance also being reflected by a relative increase in mitochondrial volume, as shown by quantitative fluorescence microscopy. These results show biosynthetic precursor production to be temporally regulated to meet phase-specific demands of eukaryal cell division.

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“…Attempts to measure Cln3 levels have been limited by its rapid degradation, causing authors to use stabilized Cln3 mutants rather than WT Cln3 (11,12). To address this concern, a recent study used a self-cleaving linker to allow a fluorescent protein to report on Cln3 translation without affecting Cln3 function or accumulation (45). These authors observed that WT cells experience a pulse of overall protein synthesis in late G1, leading to an increase in Cln3 concentration which drives cells through Start.…”

Section: Discussionmentioning

confidence: 99%

“…Previous studies have focused on the roles of Cln3, as an activator, and Whi5, as an inhibitor, in coupling passage through Start to cell size control (9,11,12,44,45). To explore whether the details of Cln3 accumulation influence cell size control, we constructed strains where the endogenous copy of CLN3 was replaced by a galactose-inducible version.…”

Section: Cell Size Control Does Not Depend On the Dynamics Of Cln3 Exmentioning

confidence: 99%

Cell size regulation in budding yeast does not depend on linear accumulation of Whi5

Barber

Amir

Murray

2020

Preprint

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Cells must couple cell cycle progress to their growth rate to restrict the spread of cell sizes present throughout a population. Linear, rather than exponential, accumulation of Whi5, was proposed to provide this coordination by causing a higher Whi5 concentration in cells born at smaller size. We tested this model using the inducible GAL1 promoter to make the Whi5 concentration independent of cell size. At an expression level that equalizes the mean cell size with that of wild-type cells, the size distributions of cells with galactose-induced Whi5 expression and wild-type cells are indistinguishable. Fluorescence microscopy confirms that the endogenous and GAL1 promoters produce different relationships between Whi5 concentration and cell volume without diminishing size control in the G1 phase. We also expressed Cln3 from the GAL1 promoter, finding that the spread in cell sizes for an asynchronous population is unaffected by this perturbation. Our findings contradict the previously proposed model for cell size control in budding yeast and demonstrate the need for a molecular mechanism that explains how cell size controls passage through Start. ClassificationBiological Sciences, Cell Biology. KeywordsWhi5, Start, cell size control, budding yeast, single-cell time-lapse microscopy. Author ContributionsFB performed the experiments, data analysis and simulations. All authors designed the experiments and wrote the manuscript. Significance StatementDespite decades of research, the question of how single cells regulate their size remains unclear. Here we demonstrate that a widely supported molecular model for the fundamental origin of size control in budding yeast is inconsistent with a set of experiments testing the model's key prediction. We therefore conclude that the problem of cell size control in budding yeast remains unsolved. This work highlights the need for rigorous testing of future models of size control in order to make progress on this fundamental question.

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Differential scaling between G1 protein production and cell size dynamics promotes commitment to the cell division cycle in budding yeast

Abstract: Differential scaling between G1 protein production and cell size dynamics promotes commitment to the cell division cycle in budding yeast.

Cited by 73 publications

References 81 publications

Mass measurements of polyploid lymphocytes reveal that growth is not size limited but depends strongly on cell cycle

Mass measurements of polyploid lymphocytes reveal that growth is not size limited but depends strongly on cell cycle

Building blocks are synthesized on demand during the yeast cell cycle

Cell size regulation in budding yeast does not depend on linear accumulation of Whi5

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