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Kukhtevich, I.

Publications and source records attributed to Kukhtevich, I..

4 recordsLinked to original sources

Swe1 delays cell cycle progression during impaired mitochondrial inheritance to promote mitochondrial homeostasis

Mitochondrial homeostasis is essential for cellular health. To maintain mitochondrial homeostasis in proliferating cells, mitochondrial inheritance to daughter cells must be tightly coordinated with growth and cell division. In budding yeast, mitochondrial inheritance is ensured by active transport into the growing bud. Although inheritance defects are well known to affect cell physiology, the regulatory mechanisms linking mitochondrial inheritance to cell cycle control remain unclear. Here, we use the inheritance defect of cells depleted of the mitochondrial inheritance adaptor Mmr1 to investigate this coordination. Time-resolved analysis of more than 8,000 budding events revealed that budding duration is directly linked to the time of mitochondrial inheritance. To understand the underlying mechanism, we performed a genetic interaction screen using the auxin-inducible degron library. Strains depleted of cell cycle regulators, particularly those involved in mitotic progression, were highly enriched in clones with synthetic growth defects. Among these, Swe1, the bud morphogenesis checkpoint kinase, emerged as a key candidate affecting budding duration in response to impaired mitochondrial inheritance. Co-depletion of Mmr1 and Swe1 strongly reduces the mitochondrial inheritance-dependent lengthening of budding duration, leading to a large fraction of daughter cells with insufficient mitochondrial content. Thus, Swe1-mediated mitotic delay contributes to slowing cell cycle progression in response to defective mitochondrial inheritance. We suggest calling this regulatory loop MIBA: mitochondrial inheritance-dependent budding adaptation. Together, our findings reveal a mechanism by which cells couple mitochondrial inheritance to cell cycle progression and highlight the importance of this coupling for mitochondrial homeostasis across the population.

cell biology↗

The origin of septin ring size control in budding yeast

The size of organelles and cellular structures needs to be tightly controlled and adjusted to the overall cell size to ensure cell function. A prominent example of a self-assembly process forming a structure whose size scales with cell size is Cdc42-driven cell polarization and subsequent septin ring formation in Saccharomyces cerevisiae. Despite extensive research, the mechanisms that determine Cdc42 cluster and septin ring size are still unclear. Combining computational modeling, live-cell imaging and genetic manipulations, we show here that positive feedback in the polarization pathway, together with the amount of polarity proteins increasing with cell size, can account for the increase of the Cdc42 cluster area with cell size, and as a consequence also the scaling of the septin ring diameter. We demonstrate that in bni1{Delta} cells, which have a larger septin ring compared to wild-type cells but a similarly sized Cdc42-GTP cluster, disruption of F-actin cable assembly and polarization toward the bud site results in diffuse exocytosis, causing septin ring enlargement. Furthermore, we find that in cells with impaired negative feedback in the polarization pathway, the Cdc42-GTP cluster area expands compared to wild-type cells, while the septin ring diameter remains mostly unchanged. This decoupling can be partially explained by a high recruitment rate of septin via polarity factors. Taken together, we provide insights into the origin of septin ring size control, in particular the scaling with cell volume, by integrating new experimental findings and mechanistic models of budding yeast polarization.

cell biology↗

Gal1 repression memory in budding yeast

Cells must continuously adapt to changing environments and, thus, have evolved mechanisms allowing them to respond to repeated stimuli. For example, faster gene induction upon a repeated stimulus aids adaptation - a process known as reinduction memory. However, whether such a memory exists for gene repression is unclear. Here, we studied gene repression across repeated carbon source shifts in over 2,500 single Saccharomyces cerevisiae cells. By monitoring the expression of a carbon source-responsive gene, galactokinase 1 (Gal1), and mathematical modeling, we discovered repression memory at the population and single-cell level. Using a repressor model to estimate single-cell repression parameters, we show that repression memory is due to a shortened repression delay, the estimated time gap between carbon source shift and Gal1 expression termination, upon the repeated carbon source shift. Additionally, we show that cells lacking Elp6 display a gain-of-repression-memory phenotype characterized by a stronger decrease in repression delay between two consecutive carbon source shifts. Collectively, our study provides the first quantitative description of repression memory in single cells.

systems biology↗

Longitudinal single cell transcriptomics reveals Krt8+ alveolar epithelial progenitors in lung regeneration

Lung injury activates quiescent stem and progenitor cells to regenerate alveolar structures. The sequence and coordination of transcriptional programs during this process has largely remained elusive. Using single cell RNA-seq, we first generated a whole-organ birds-eye view on cellular dynamics and cell-cell communication networks during mouse lung regeneration from [~]30,000 cells at six timepoints. We discovered an injury-specific progenitor cell state characterized by Krt8 in flat epithelial cells covering alveolar surfaces. The number of these cells peaked during fibrogenesis in independent mouse models, as well as in human acute lung injury and fibrosis. Krt8+ progenitors featured a highly distinct connectome of receptor-ligand pairs with endothelial cells, fibroblasts, and macrophages. To sky dive into epithelial differentiation dynamics, we sequenced >30,000 sorted epithelial cells at 18 timepoints and computationally derived cell state trajectories that were validated by lineage tracing genetic reporter mice. Airway stem cells within the club cell lineage and alveolar type-2 cells underwent transcriptional convergence onto the same Krt8+ progenitor cell state, which later resolved by terminal differentiation into alveolar type-1 cells. We derived distinct transcriptional regulators as key switch points in this process and show that induction of TNF-alpha/NFkappaB, p53, and hypoxia driven gene expression programs precede a Sox4, Ctnnb1, and Wwtr1 driven switch towards alveolar type-1 cell fate. We show that epithelial cell plasticity can induce non-gradual transdifferentiation, involving intermediate progenitor cell states that may persist and promote disease if checkpoint signals for terminal differentiation are perturbed.

systems biology↗