Genome concentration limits cell growth and modulates proteome composition in Escherichia coli DOI Creative Commons
Jarno Mäkelä, Alexandros Papagiannakis, Wei-Hsiang Lin

et al.

eLife, Journal Year: 2024, Volume and Issue: 13

Published: Dec. 23, 2024

Defining the cellular factors that drive growth rate and proteome composition is essential for understanding manipulating systems. In bacteria, ribosome concentration known to be a constraining factor of cell rate, while gene usually assumed not limiting. Here, using single-molecule tracking, quantitative single-cell microscopy, modeling, we show genome dilution in Escherichia coli cells arrested DNA replication limits total RNA polymerase activity within physiological sizes across tested nutrient conditions. This rapid-onset limitation on bulk transcription results sub-linear scaling active ribosomes with size sub-exponential growth. Such downstream effects translation are near-immediately detectable nutrient-rich medium, but delayed nutrient-poor conditions, presumably due buffering activities. sequencing tandem-mass-tag mass spectrometry experiments further reveal remodels relative abundance mRNAs proteins at global level. Altogether, our findings indicate chromosome limiting modulator transcriptome E. . Experiments Caulobacter crescentus comparison eukaryotic studies identify broadly conserved concentration-dependent principles expression.

Language: Английский

CDK4/6 inhibitor-mediated cell overgrowth triggers osmotic and replication stress to promote senescence DOI Creative Commons
Lisa Crozier, Reece Foy, Rozita Adib

et al.

Molecular Cell, Journal Year: 2023, Volume and Issue: 83(22), P. 4062 - 4077.e5

Published: Nov. 1, 2023

Abnormal increases in cell size are associated with senescence and cycle exit. The mechanisms by which overgrowth primes cells to withdraw from the remain unknown. We address this question using CDK4/6 inhibitors, arrest G0/G1 licensed treat advanced HR+/HER2- breast cancer. demonstrate that CDK4/6-inhibited overgrow during G0/G1, causing p38/p53/p21-dependent withdrawal. Cell withdrawal is triggered biphasic p21 induction. first wave caused osmotic stress, leading p38- size-dependent accumulation of p21. inhibitor washout results some entering S-phase. Overgrown experience replication resulting a second promotes G2 or subsequent G1. propose levels integrate signals overgrowth-triggered stresses determine fate. This model explains how hypertrophy can drive why inhibitors have long-lasting effects patients.

Language: Английский

Citations

44
Genome homeostasis defects drive enlarged cells into senescence DOI Creative Commons
Sandhya Manohar, Marianna E. Estrada, Federico Uliana

et al.

Molecular Cell, Journal Year: 2023, Volume and Issue: 83(22), P. 4032 - 4046.e6

Published: Nov. 1, 2023

Cellular senescence refers to an irreversible state of cell-cycle arrest and plays important roles in aging cancer biology. Because is associated with increased cell size, we used reversible arrests combined growth rate modulation study how excessive affects proliferation. We find that enlarged cells upregulate p21, which limits progression. Cells re-enter the cycle encounter replication stress well tolerated physiologically sized but causes severe DNA damage cells, ultimately resulting mitotic failure permanent withdrawal. demonstrate fail recruit 53BP1 other non-homologous end joining (NHEJ) machinery sites robustly initiate damage-dependent p53 signaling, rendering them highly sensitive genotoxic stress. propose impaired response primes for persistent replication-acquired damage, leading division exit.

Language: Английский

Citations

38
Oncogenic signals prime cancer cells for toxic cell overgrowth during a G1 cell cycle arrest DOI Creative Commons
Reece Foy, Lisa Crozier,

Aanchal Udaynath Pareri

et al.

Molecular Cell, Journal Year: 2023, Volume and Issue: 83(22), P. 4047 - 4061.e6

Published: Nov. 1, 2023

CDK4/6 inhibitors are remarkable anti-cancer drugs that can arrest tumor cells in G1 and induce their senescence while causing only relatively mild toxicities healthy tissues. How they achieve this mechanistically is unclear. We show here specifically vulnerable to inhibition because during the arrest, oncogenic signals drive toxic cell overgrowth. This overgrowth causes permanent cycle withdrawal by either preventing progression from or inducing genotoxic damage subsequent S-phase mitosis. Inhibiting reverting converge onto mTOR rescue excessive growth, DNA damage, exit cancer cells. Conversely, non-transformed these phenotypes sensitize inhibition. Together, demonstrates growth a synthetic lethal combination exploited tumor-specific toxicity.

Language: Английский

Citations

34
Mechanisms of sensitivity and resistance to CDK4/CDK6 inhibitors in hormone receptor-positive breast cancer treatment DOI Creative Commons
Antonino Glaviano, Seth A. Wander, Richard D. Baird

et al.

Drug Resistance Updates, Journal Year: 2024, Volume and Issue: 76, P. 101103 - 101103

Published: June 25, 2024

Cell cycle dysregulation is a hallmark of cancer that promotes eccessive cell division. Cyclin-dependent kinase 4 (CDK4) and cyclin-dependent 6 (CDK6) are key molecules in the G1-to-S phase transition crucial for onset, survival, progression breast (BC). Small-molecule CDK4/CDK6 inhibitors (CDK4/6i) block phosphorylation tumor suppressor Rb thus restrain susceptible BC cells G1 phase. Three CDK4/6i approved first-line treatment patients with advanced/metastatic hormone receptor-positive (HR

Language: Английский

Citations

14
The search for CDK4/6 inhibitor biomarkers has been hampered by inappropriate proliferation assays DOI Creative Commons
Reece Foy,

Kah Xin Lew,

Adrian T. Saurin

et al.

npj Breast Cancer, Journal Year: 2024, Volume and Issue: 10(1)

Published: March 4, 2024

CDK4/6 inhibitors are effective at treating advanced HR+ /HER2- breast cancer, however biomarkers that can predict response urgently needed. We demonstrate here previous large-scale screens designed to identify which tumour types or genotypes most sensitive have misrepresented the responsive cell lines because of a reliance on metabolic proliferation assays. CDK4/6-inhibited cells arrest in G1 but continue grow size, thereby producing more mitochondria. show this growth obscures using ATP-based assays not if DNA-based used instead. Furthermore, lymphoma lines, previously identified as sensitive, simply appear respond best they fail overgrow during arrest. Similarly, inhibitor abemaciclib appears inhibit better than palbociclib it also restricts cellular overgrowth through off-target effects. DepMap analysis screening data reliable assay types, demonstrates palbociclib-sensitive Cyclin D1, CDK4 and CDK6 knockout/knockdown, whereas palbociclib-resistant E1, CDK2 SKP2 knockout/knockdown. Potential increased expression CCND1 RB1, reduced CCNE1 CDKN2A. Probing with similar from fails reveal these associations. Together, why inhibitors, any other anti-cancer drugs cycle permit continued growth, must now be re-screened against wide-range an appropriate assay. This would help inform clinical trials much needed response.

Language: Английский

Citations

9
Genome dilution by cell growth drives starvation-like proteome remodeling in mammalian and yeast cells DOI
Michael C. Lanz, Shuyuan Zhang, Matthew P. Swaffer

et al.

Nature Structural & Molecular Biology, Journal Year: 2024, Volume and Issue: unknown

Published: July 24, 2024

Language: Английский

Citations

9
Structural basis of Cdk7 activation by dual T-loop phosphorylation DOI Creative Commons
Robert Düster, K. Anand, Sophie C. Binder

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: Aug. 3, 2024

Abstract Cyclin-dependent kinase 7 (Cdk7) is required in cell-cycle and transcriptional regulation owing to its function as both a CDK-activating (CAK) part of transcription factor TFIIH. Cdk7 forms active complexes by associating with Cyclin H Mat1, regulated two phosphorylations the activation segment (T loop): canonical activating modification at T170 another S164. Here we report crystal structure human Cdk7/Cyclin H/Mat1 complex containing T-loop phosphorylations. Whereas pT170 coordinates basic residues conserved other CDKs, pS164 nucleates an arginine network unique ternary complex, involving all three subunits. We identify differential dependencies activity substrate recognition on individual CAK unaffected phosphorylation, whereas towards non-CDK substrates increased several-fold phosphorylation. Moreover, dual phosphorylation stimulates multisite RNA polymerase II (RNAPII) carboxy-terminal domain (CTD) SPT5 repeat (CTR) region. In cells, two-step process wherein S164 precedes, may prime, Thus, can regulate through multiple mechanisms, supporting tripartite formation possibly influencing processivity, while enhances key substrates.

Language: Английский

Citations

9
Delineation of proteome changes driven by cell size and growth rate DOI Creative Commons
Evgeny Zatulovskiy, Michael C. Lanz, Shuyuan Zhang

et al.

Frontiers in Cell and Developmental Biology, Journal Year: 2022, Volume and Issue: 10

Published: Sept. 5, 2022

Increasing cell size drives changes to the proteome, which affects physiology. As increases, some proteins become more concentrated while others are diluted. a result, state of continuously with increasing size. In addition these proteomic changes, large cells have lower growth rate (protein synthesis per unit volume). That both cell's proteome and change suggests they may be interdependent. To test this, we used quantitative mass spectrometry measure how in response mTOR inhibitor rapamycin, decreases cellular has only minimal effect on We found that inhibition, cell, remodel similar ways. This many effects mediated by size-dependent slowdown rate. For example, previously reported expression senescence markers could reflect declining rather than its

Language: Английский

Citations

23
Eukaryotic cell size regulation and its implications for cellular function and dysfunction DOI
Yagya Chadha, Arohi Khurana, Kurt M. Schmoller

et al.

Physiological Reviews, Journal Year: 2024, Volume and Issue: 104(4), P. 1679 - 1717

Published: June 20, 2024

Depending on cell type, environmental inputs, and disease, the cells in human body can have widely different sizes. In recent years, it has become clear that size is a major regulator of function. However, we are only beginning to understand how optimization function determines given cell’s optimal size. Here, review currently known control strategies eukaryotic intricate link intracellular biomolecular scaling, organelle homeostasis, cycle progression. We detail size-dependent regulation early development impact differentiation. Given importance for normal cellular physiology, must account changing conditions. describe sense stimuli, such as nutrient availability, accordingly adapt their by regulating growth Moreover, discuss correlation pathological states with misregulation long time this was considered downstream consequence dysfunction. newer studies reveal reversed causality, misregulated leading pathophysiological phenotypes senescence aging. summary, highlight important roles dysfunction, which could implications both diagnostics treatment clinic.

Language: Английский

Citations

5
Cellular enlargement - A new hallmark of aging? DOI Creative Commons

Daniel Davies,

Kim van den Handel,

Soham Bharadwaj

et al.

Frontiers in Cell and Developmental Biology, Journal Year: 2022, Volume and Issue: 10

Published: Nov. 10, 2022

Years of important research has revealed that cells heavily invest in regulating their size. Nevertheless, it remained unclear why accurate size control is so important. Our recent study using hematopoietic stem (HSCs) vivo indicates cellular enlargement causally associated with aging. Here, we present an overview these findings and implications. Furthermore, performed a broad literature analysis to evaluate the potential as new aging hallmark examine its connection previously described hallmarks. Finally, highlight interesting work presenting correlation between cell age-related diseases. Taken together, found mounting evidence linking Therefore, encourage researchers from seemingly unrelated areas take fresh look at data perspective

Language: Английский

Citations

19