Quantitative assessment of changes in cell growth, size and morphology during telomere-initiated cellular senescence in Saccharomyces cerevisiae

Ghanem, Neda Z; Malla, Shubha R.L.; Araki, Naoko; Lewis, L. Kevin

doi:10.1016/j.yexcr.2019.05.005

Cited by 3 publications

(6 citation statements)

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“…To test other components of NHEJ, rescue of yku70 cells that also had DNA Ligase IV ( DNL4 ) inactivated was assessed and found to be similar to that of yku70 single mutants ( Figure 7, A and B ). RAD52 is critical for the repair of DSBs by the HR pathway and is known to affect telomere shortening rates in cells undergoing senescence ( Lundblad and Blackburn 1993 ; Le et al 1999 ; Gatbonton et al 2006 ; Gao et al 2010 ; Becerra et al 2012 ; Teixeira 2013 ; Ghanem et al 2019 ). Growth of yku70 rad52 double mutants was strongly enhanced at 37°C when either Est2 or TLC1 RNA was overexpressed, though colony growth was modestly reduced compared to rescued yku70 single mutants ( Figure 7, A and B ).…”

Section: Resultsmentioning

confidence: 99%

“…For example, telomerase-deficient est2 or tlc1 mutants experience telomere shortening and undergo replicative senescence when grown in culture, dividing approximately 70 times before growth capability is lost. However, telomerase-deficient cells can only undergo approximately 40 cell divisions when RAD52 is coinactivated ( Lundblad and Blackburn 1993 ; Khadaroo et al 2009 ; Becerra et al 2012 ; Ghanem et al 2019 ). Our results suggest that a similar process occurs when yku70 cells undergo telomere shortening at 37°C, i.e.…”

Section: Discussionmentioning

confidence: 99%

“…This strain was propagated on synthetic galactose plates without uracil and then streaked to glucose complete plates to initiate and monitor senescence. Repetitive picking and streaking of individual colonies from one plate to another result in senescence (loss of growth capability) that is visible on the 4th streak if medium-sized colonies are picked each time and on the 5th streak plate when small colonies are picked ( Becerra et al 2012 ; Ghanem et al 2019 ). Counting cells within the small colonies used in this study indicated that each had undergone approximately 18–19 generations before being picked and streaked to a new plate.…”

Section: Methodsmentioning

confidence: 99%

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Suppression of telomere capping defects of Saccharomyces cerevisiae yku70 and yku80 mutants by telomerase

Holland

Sanderson

Titus

et al. 2021

G3 Genes|Genomes|Genetics

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The Ku complex performs multiple functions inside eukaryotic cells, including protection of chromosomal DNA ends from degradation and fusion events, recruitment of telomerase, and repair of double-strand breaks (DSBs). Inactivation of Ku complex genes YKU70 or YKU80 in cells of the yeast S. cerevisiae gives rise to mutants that exhibit shortened telomeres and temperature-sensitive growth. In this study we have investigated the mechanism by which overexpression of telomerase suppresses the temperature sensitivity of yku mutants. Viability of yku cells was restored by overexpression of the Est2 reverse transcriptase and TLC1 RNA template subunits of telomerase, but not the Est1 or Est3 proteins. Overexpression of other telomerase- and telomere-associated proteins (Cdc13, Stn1, Ten1, Rif1, Rif2, Sir3, Sir4) did not suppress the growth defects of yku70 cells. Mechanistic features of suppression were assessed using several TLC1 RNA deletion derivatives and Est2 enzyme mutants. Supraphysiological levels of three catalytically inactive reverse transcriptase mutants (Est2-D530A, Est2-D670A and Est2-D671A) suppressed the loss of viability as efficiently as the wildtype Est2 protein, without inducing cell senescence. Roles of proteins regulating telomere length were also determined. The results support a model in which chromosomes in yku mutants are stabilized via a replication-independent mechanism involving structural reinforcement of protective telomere cap structures.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Suppression of telomere capping defects of Saccharomyces cerevisiae yku70 and yku80 mutants by telomerase

Holland

Sanderson

Titus

et al. 2021

G3 Genes|Genomes|Genetics

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show abstract

“…It consists of the telomerase reverse transcriptase of telomerase RNA (TR), which serves as a template for synthesis (Greider and Blackburn, 1989 ; Feng et al, 1995 ; Lingner et al, 1997 ). Damage due to inactivation of telomerase results in a DNA damage checkpoint response, which leads to changes in size, morphology, and cellular senescence in Saccharomyces cerevisiae ( S. cerevisiae ) cells after ~60–70 cell cycles (Ghanem et al, 2019 ).…”

Section: General Aspects Of Agingmentioning

confidence: 99%

“…Besides rDNA stability and copy number, telomere length has an impact on yeast cell morphology. Telomere senescence resulted in an increase in the diameters of unbudded cells from 5.2 to 8.2 μm and of budded cells from 6.3 to 10.1 μm, indicating an increase of 58 and 60%, respectively (Ghanem et al, 2019 ). This change in cell diameter negatively influences the optimal ratio of DNA:cytoplasm, supporting the observed non-optimal cell function (Neurohr et al, 2019 ).…”

Section: Aging Effects In Yeast Cultures Under Industrial Processingmentioning

confidence: 99%

Understanding the Impact of Industrial Stress Conditions on Replicative Aging in Saccharomyces cerevisiae

Eigenfeld

Kerpes

Becker

2021

Front. Fungal Biol.

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In yeast, aging is widely understood as the decline of physiological function and the decreasing ability to adapt to environmental changes. Saccharomyces cerevisiae has become an important model organism for the investigation of these processes. Yeast is used in industrial processes (beer and wine production), and several stress conditions can influence its intracellular aging processes. The aim of this review is to summarize the current knowledge on applied stress conditions, such as osmotic pressure, primary metabolites (e.g., ethanol), low pH, oxidative stress, heat on aging indicators, age-related physiological changes, and yeast longevity. There is clear evidence that yeast cells are exposed to many stressors influencing viability and vitality, leading to an age-related shift in age distribution. Currently, there is a lack of rapid, non-invasive methods allowing the investigation of aspects of yeast aging in real time on a single-cell basis using the high-throughput approach. Methods such as micromanipulation, centrifugal elutriator, or biotinylation do not provide real-time information on age distributions in industrial processes. In contrast, innovative approaches, such as non-invasive fluorescence coupled flow cytometry intended for high-throughput measurements, could be promising for determining the replicative age of yeast cells in fermentation and its impact on industrial stress conditions.

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Deficiency in homologous recombination is associated with changes in cell cycling and morphology in Saccharomyces cerevisiae

Holland

Weis

England

et al. 2023

Experimental Cell Research

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Quantitative assessment of changes in cell growth, size and morphology during telomere-initiated cellular senescence in Saccharomyces cerevisiae

Cited by 3 publications

References 73 publications

Suppression of telomere capping defects of Saccharomyces cerevisiae yku70 and yku80 mutants by telomerase

Suppression of telomere capping defects of Saccharomyces cerevisiae yku70 and yku80 mutants by telomerase

Understanding the Impact of Industrial Stress Conditions on Replicative Aging in Saccharomyces cerevisiae

Deficiency in homologous recombination is associated with changes in cell cycling and morphology in Saccharomyces cerevisiae

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