bims-pideca Biomed News
on Class IA PI3K signalling in development and cancer
Issue of 2023‒11‒12
sixteen papers selected by
Ralitsa Radostinova Madsen, MRC-PPU
  1. Molecular characterization of 13 patients with PIK3CA-related overgrowth spectrum using a targeted deep sequencing approach.
  2. CBAP regulates the function of Akt-associated TSC protein complexes to modulate mTORC1 signaling.
  3. Flow cytometry-based quantification of genome editing efficiency in human cell lines using the L1CAM gene.
  4. Disrupting cellular memory to overcome drug resistance.
  5. A statistical framework for differential pseudotime analysis with multiple single-cell RNA-seq samples.
  6. Neurodevelopmental disorders and cancer networks share pathways, but differ in mechanisms, signaling strength, and outcome.
  7. The Reactome Pathway Knowledgebase 2024.
  8. Deubiquitinases in cancer.
  9. Scalable single-cell profiling of chromatin modifications with sciCUT&Tag.
  10. Preliminary results of the European multicentric phase III trial regarding sirolimus in slow-flow vascular malformations.
  11. Pharmacologic activation of a compensatory integrated stress response kinase promotes mitochondrial remodeling in PERK-deficient cells.
  12. Tissue-specific Cre driver mice to study vascular diseases.
  13. Selective Inhibition of mTORC1 Signaling Supports the Development and Maintenance of Pluripotency.
  14. Towards inclusive and sustainable scientific meetings.
  15. Editorial: Single cell dynamics and cell cycle length variation.
  16. RNA polymerase II dynamics and mRNA stability feedback scale mRNA amounts with cell size.
  1. Am J Med Genet A. 2023 Nov 10.
    Molecular characterization of 13 patients with PIK3CA-related overgrowth spectrum using a targeted deep sequencing approach.
    Care4Rare Canada Consortium
      Activating variants in the PIK3CA gene cause a heterogeneous spectrum of disorders that involve congenital or early-onset segmental/focal overgrowth, now referred to as PIK3CA-related overgrowth spectrum (PROS). Historically, the clinical diagnoses of patients with PROS included a range of distinct syndromes, including CLOVES syndrome, dysplastic megalencephaly, hemimegalencephaly, focal cortical dysplasia, Klippel-Trenaunay syndrome, CLAPO syndrome, fibroadipose hyperplasia or overgrowth, hemihyperplasia multiple lipomatosis, and megalencephaly capillary malformation-polymicrogyria (MCAP) syndrome. MCAP is a sporadic overgrowth disorder that exhibits core features of progressive megalencephaly, vascular malformations, distal limb malformations, cortical brain malformations, and connective tissue dysplasia. In 2012, our research group contributed to the identification of predominantly mosaic, gain-of-function variants in PIK3CA as an underlying genetic cause of the syndrome. Mosaic variants are technically more difficult to detect and require implementation of more sensitive sequencing technologies and less stringent variant calling algorithms. In this study, we demonstrated the utility of deep sequencing using the Illumina TruSight Oncology 500 (TSO500) sequencing panel in identifying variants with low allele fractions in a series of patients with PROS and suspected mosaicism: pathogenic, mosaic PIK3CA variants were identified in all 13 individuals, including 6 positive controls. This study highlights the importance of screening for low-level mosaic variants in PROS patients. The use of targeted panels with deep sequencing in clinical genetic testing laboratories would improve diagnostic yield and accuracy within this patient population.
    Keywords:  PIK3CA; PROS; mosaicism; overgrowth syndrome; rare genetic disease
    DOI:  https://doi.org/10.1002/ajmg.a.63466
  2. J Biol Chem. 2023 Nov 08. pii: S0021-9258(23)02483-3. [Epub ahead of print] 105455
    CBAP regulates the function of Akt-associated TSC protein complexes to modulate mTORC1 signaling.
    Wei-Ting Liao, Yun-Jung Chiang, Hsin-Fang Yang-Yen, Li-Chung Hsu, Zee-Fen Chang, Jeffrey Jy Yen.
      The Akt-Rheb-mTORC1 pathway plays a crucial role in regulating cell growth, but the mechanisms underlying the activation of Rheb-mTORC1 by Akt remain unclear. In our previous study, we found that CBAP was highly expressed in human T-ALL cells and primary tumors, and its deficiency led to reduced phosphorylation of TSC2/S6K1 signaling proteins, as well as impaired cell proliferation and leukemogenicity. We also demonstrated that CBAP was required for Akt-mediated TSC2 phosphorylation in vitro. In response to insulin, CBAP was also necessary for phosphorylation of TSC2/S6K1 and the dissociation of TSC2 from the lysosomal membrane. Here we report that CBAP interacts with AKT and TSC2, and knockout of CBAP or serum starvation leads to an increase in TSC1 in the Akt/TSC2 immunoprecipitation complexes. Lysosomal-anchored CBAP was found to override serum starvation and promote S6K1 and 4EBP1 phosphorylation and c-Myc expression in a TSC2-dependent manner. Additionally, recombinant CBAP inhibited the GAP activity of TSC2 complexes in vitro, leading to increased Rheb-GTP loading, likely due to the competition between TSC1 and CBAP for binding to the HBD domain of TSC2. Overexpression of the N26 region of CBAP, which is crucial for binding to TSC2, resulted in a decrease in mTORC1 signaling and an increase in TSC1 association with the TSC2/AKT complex, ultimately leading to increased GAP activity toward Rheb and impaired cell proliferation. Thus, we propose that CBAP can modulate the stability of TSC1-TSC2 as well as promote translocation of TSC1/TSC2 complexes away from lysosomes to regulate Rheb-mTORC1 signaling.
    Keywords:  Akt; Rheb; cell growth; mTORC1 activation; small GTPase; tumor cell biology
    DOI:  https://doi.org/10.1016/j.jbc.2023.105455
  3. PLoS One. 2023 ;18(11): e0294146
    Flow cytometry-based quantification of genome editing efficiency in human cell lines using the L1CAM gene.
    Muhammad Nazmul Hasan, Toshinori Hyodo, Mrityunjoy Biswas, Md Lutfur Rahman, Yuko Mihara, Sivasundaram Karnan, Akinobu Ota, Shinobu Tsuzuki, Yoshitaka Hosokawa, Hiroyuki Konishi.
      CRISPR/Cas9 is a powerful genome editing system that has remarkably facilitated gene knockout and targeted knock-in. To accelerate the practical use of CRISPR/Cas9, however, it remains crucial to improve the efficiency, precision, and specificity of genome editing, particularly targeted knock-in, achieved with this system. To improve genome editing efficiency, researchers should first have a molecular assay that allows sensitive monitoring of genome editing events with simple procedures. In the current study, we demonstrate that genome editing events occurring in L1CAM, an X-chromosome gene encoding a cell surface protein, can be readily monitored using flow cytometry (FCM) in multiple human cell lines including neuroblastoma cell lines. The abrogation of L1CAM was efficiently achieved using Cas9 nucleases which disrupt exons encoding the L1CAM extracellular domain, and was easily detected by FCM using anti-L1CAM antibodies. Notably, L1CAM-abrogated cells could be quantified by FCM in four days after transfection with a Cas9 nuclease, which is much faster than an established assay based on the PIGA gene. In addition, the L1CAM-based assay allowed us to measure the efficiency of targeted knock-in (correction of L1CAM mutations) accomplished through different strategies, including a Cas9 nuclease-mediated method, tandem paired nicking, and prime editing. Our L1CAM-based assay using FCM enables rapid and sensitive quantification of genome editing efficiencies and will thereby help researchers improve genome editing technologies.
    DOI:  https://doi.org/10.1371/journal.pone.0294146
  4. Nat Commun. 2023 Nov 06. 14(1): 7130
    Disrupting cellular memory to overcome drug resistance.
    Guillaume Harmange, Raúl A Reyes Hueros, Dylan L Schaff, Benjamin Emert, Michael Saint-Antoine, Laura C Kim, Zijian Niu, Shivani Nellore, Mitchell E Fane, Gretchen M Alicea, Ashani T Weeraratna, M Celeste Simon, Abhyudai Singh, Sydney M Shaffer.
      Gene expression states persist for varying lengths of time at the single-cell level, a phenomenon known as gene expression memory. When cells switch states, losing memory of their prior state, this transition can occur in the absence of genetic changes. However, we lack robust methods to find regulators of memory or track state switching. Here, we develop a lineage tracing-based technique to quantify memory and identify cells that switch states. Applied to melanoma cells without therapy, we quantify long-lived fluctuations in gene expression that are predictive of later resistance to targeted therapy. We also identify the PI3K and TGF-β pathways as state switching modulators. We propose a pretreatment model, first applying a PI3K inhibitor to modulate gene expression states, then applying targeted therapy, which leads to less resistance than targeted therapy alone. Together, we present a method for finding modulators of gene expression memory and their associated cell fates.
    DOI:  https://doi.org/10.1038/s41467-023-41811-8
  5. Nat Commun. 2023 Nov 10. 14(1): 7286
    A statistical framework for differential pseudotime analysis with multiple single-cell RNA-seq samples.
    Wenpin Hou, Zhicheng Ji, Zeyu Chen, E John Wherry, Stephanie C Hicks, Hongkai Ji.
      Pseudotime analysis with single-cell RNA-sequencing (scRNA-seq) data has been widely used to study dynamic gene regulatory programs along continuous biological processes. While many methods have been developed to infer the pseudotemporal trajectories of cells within a biological sample, it remains a challenge to compare pseudotemporal patterns with multiple samples (or replicates) across different experimental conditions. Here, we introduce Lamian, a comprehensive and statistically-rigorous computational framework for differential multi-sample pseudotime analysis. Lamian can be used to identify changes in a biological process associated with sample covariates, such as different biological conditions while adjusting for batch effects, and to detect changes in gene expression, cell density, and topology of a pseudotemporal trajectory. Unlike existing methods that ignore sample variability, Lamian draws statistical inference after accounting for cross-sample variability and hence substantially reduces sample-specific false discoveries that are not generalizable to new samples. Using both real scRNA-seq and simulation data, including an analysis of differential immune response programs between COVID-19 patients with different disease severity levels, we demonstrate the advantages of Lamian in decoding cellular gene expression programs in continuous biological processes.
    DOI:  https://doi.org/10.1038/s41467-023-42841-y
  6. NPJ Genom Med. 2023 Nov 04. 8(1): 37
    Neurodevelopmental disorders and cancer networks share pathways, but differ in mechanisms, signaling strength, and outcome.
    Bengi Ruken Yavuz, M Kaan Arici, Habibe Cansu Demirel, Chung-Jung Tsai, Hyunbum Jang, Ruth Nussinov, Nurcan Tuncbag.
      Epidemiological studies suggest that individuals with neurodevelopmental disorders (NDDs) are more prone to develop certain types of cancer. Notably, however, the case statistics can be impacted by late discovery of cancer in individuals afflicted with NDDs, such as intellectual disorders, autism, and schizophrenia, which may bias the numbers. As to NDD-associated mutations, in most cases, they are germline while cancer mutations are sporadic, emerging during life. However, somatic mosaicism can spur NDDs, and cancer-related mutations can be germline. NDDs and cancer share proteins, pathways, and mutations. Here we ask (i) exactly which features they share, and (ii) how, despite their commonalities, they differ in clinical outcomes. To tackle these questions, we employed a statistical framework followed by network analysis. Our thorough exploration of the mutations, reconstructed disease-specific networks, pathways, and transcriptome levels and profiles of autism spectrum disorder (ASD) and cancers, point to signaling strength as the key factor: strong signaling promotes cell proliferation in cancer, and weaker (moderate) signaling impacts differentiation in ASD. Thus, we suggest that signaling strength, not activating mutations, can decide clinical outcome.
    DOI:  https://doi.org/10.1038/s41525-023-00377-6
  7. Nucleic Acids Res. 2023 Nov 06. pii: gkad1025. [Epub ahead of print]
    The Reactome Pathway Knowledgebase 2024.
    Marija Milacic, Deidre Beavers, Patrick Conley, Chuqiao Gong, Marc Gillespie, Johannes Griss, Robin Haw, Bijay Jassal, Lisa Matthews, Bruce May, Robert Petryszak, Eliot Ragueneau, Karen Rothfels, Cristoffer Sevilla, Veronica Shamovsky, Ralf Stephan, Krishna Tiwari, Thawfeek Varusai, Joel Weiser, Adam Wright, Guanming Wu, Lincoln Stein, Henning Hermjakob, Peter D'Eustachio.
      The Reactome Knowledgebase (https://reactome.org), an Elixir and GCBR core biological data resource, provides manually curated molecular details of a broad range of normal and disease-related biological processes. Processes are annotated as an ordered network of molecular transformations in a single consistent data model. Reactome thus functions both as a digital archive of manually curated human biological processes and as a tool for discovering functional relationships in data such as gene expression profiles or somatic mutation catalogs from tumor cells. Here we review progress towards annotation of the entire human proteome, targeted annotation of disease-causing genetic variants of proteins and of small-molecule drugs in a pathway context, and towards supporting explicit annotation of cell- and tissue-specific pathways. Finally, we briefly discuss issues involved in making Reactome more fully interoperable with other related resources such as the Gene Ontology and maintaining the resulting community resource network.
    DOI:  https://doi.org/10.1093/nar/gkad1025
  8. Nat Rev Cancer. 2023 Nov 07.
    Deubiquitinases in cancer.
    Grant Dewson, Pieter J A Eichhorn, David Komander.
      Ubiquitination is an essential regulator of most, if not all, signalling pathways, and defects in cellular signalling are central to cancer initiation, progression and, eventually, metastasis. The attachment of ubiquitin signals by E3 ubiquitin ligases is directly opposed by the action of approximately 100 deubiquitinating enzymes (DUBs) in humans. Together, DUBs and E3 ligases coordinate ubiquitin signalling by providing selectivity for different substrates and/or ubiquitin signals. The balance between ubiquitination and deubiquitination is exquisitely controlled to ensure properly coordinated proteostasis and response to cellular stimuli and stressors. Not surprisingly, then, DUBs have been associated with all hallmarks of cancer. These relationships are often complex and multifaceted, highlighted by the implication of multiple DUBs in certain hallmarks and by the impact of individual DUBs on multiple cancer-associated pathways, sometimes with contrasting cancer-promoting and cancer-inhibiting activities, depending on context and tumour type. Although it is still understudied, the ever-growing knowledge of DUB function in cancer physiology will eventually identify DUBs that warrant specific inhibition or activation, both of which are now feasible. An integrated appreciation of the physiological consequences of DUB modulation in relevant cancer models will eventually lead to the identification of patient populations that will most likely benefit from DUB-targeted therapies.
    DOI:  https://doi.org/10.1038/s41568-023-00633-y
  9. Nat Protoc. 2023 Nov 07.
    Scalable single-cell profiling of chromatin modifications with sciCUT&Tag.
    Derek H Janssens, Jacob E Greene, Steven J Wu, Christine A Codomo, Samuel S Minot, Scott N Furlan, Kami Ahmad, Steven Henikoff.
      Cleavage under targets and tagmentation (CUT&Tag) is an antibody-directed in situ chromatin profiling strategy that is rapidly replacing immune precipitation-based methods, such as chromatin immunoprecipitation-sequencing. The efficiency of the method enables chromatin profiling in single cells but is limited by the numbers of cells that can be profiled. Here, we describe a combinatorial barcoding strategy for CUT&Tag that harnesses a nanowell dispenser for simple, high-resolution, high-throughput, single-cell chromatin profiling. In this single-cell combinatorial indexing CUT&Tag (sciCUT&Tag) protocol, lightly cross-linked nuclei are bound to magnetic beads and incubated with primary and secondary antibodies in bulk and then arrayed in a 96-well plate for a first round of cellular indexing by antibody-directed Tn5 tagmentation. The sample is then repooled, mixed and arrayed across 5,184 nanowells at a density of 12-24 nuclei per well for a second round of cellular indexing during PCR amplification of the sequencing-ready library. This protocol can be completed in 1.5 days by a research technician, and we illustrate the optimized protocol by profiling histone modifications associated with developmental gene repression (H3K27me3) as well as transcriptional activation (H3K4me1-2-3) in human peripheral blood mononuclear cells and use single-nucleotide polymorphisms to facilitate collision removal. We have also used sciCUT&Tag for simultaneous profiling of multiple chromatin epitopes in single cells. The reduced cost, improved resolution and scalability of sciCUT&Tag make it an attractive platform to profile chromatin features in single cells.
    DOI:  https://doi.org/10.1038/s41596-023-00905-9
  10. JCI Insight. 2023 Nov 08. pii: e173095. [Epub ahead of print]8(21):
    Preliminary results of the European multicentric phase III trial regarding sirolimus in slow-flow vascular malformations.
    Emmanuel Seront, An Van Damme, Catherine Legrand, Annouk Bisdorff-Bresson, Philippe Orcel, Thomas Funck-Brentano, Marie-Antoinette Sevestre, Anne Dompmartin, Isabelle Quere, Pascal Brouillard, Nicole Revencu, Martina De Bortoli, Frank Hammer, Philippe Clapuyt, Dana Dumitriu, Miikka Vikkula, Laurence M Boon.
      BACKGROUNDSlow-flow vascular malformations frequently harbor activating mutations in the PI3K/AKT/mTOR cascade. Phase II trials pinpointed sirolimus effectiveness as a drug therapy. Efficacy and safety of sirolimus thus need to be evaluated in large prospective phase III trials.METHODSThe Vascular Anomaly-Sirolimus-Europe (VASE) trial, initiated in 2016, is a large multicentric prospective phase III trial (EudraCT 2015-001703-32), which evaluates efficacy and safety of sirolimus for 2 years in pediatric and adult patients with symptomatic slow-flow vascular malformations. In this interim analysis, we studied all patients enrolled up to October 2021 who received sirolimus for 12 or more months or who prematurely stopped the treatment.RESULTSThirty-one pediatric and 101 adult patients were included in this analysis; 107 completed 12 or more months of sirolimus, including 61 who were treated for the whole 2-year period. Sirolimus resulted in a clinical improvement in 85% of patients. The efficacy appeared within the first month for the majority of them. Grade 3-4 adverse events were observed in 24 (18%) patients; all resolved after treatment interruption/arrest. Sirolimus increased feasibility of surgery or sclerotherapy in 20 (15%) patients initially deemed unsuitable for intervention. Among the 61 patients who completed the 2-year treatment, 33 (54%) reported a recurrence of symptoms after a median follow-up of 13 months after sirolimus arrest. While there was no difference in efficacy, clinical improvement was faster but subsided more rapidly in PIK3CA-mutated (n = 24) compared with TIE2-mutated (n = 19) patients.CONCLUSIONSirolimus has a high efficacy and good tolerance in treatment of slow-flow vascular malformations in children and adults.TRIAL REGISTRATIONClinicalTrials.gov NCT02638389 and EudraCT 2015-001703-32.FUNDINGThe Fonds de la Recherche Scientifique (FNRS grants T.0247.19, P.C005.22, T.0146.16, and P.C013.20), the Fund Generet managed by the King Baudouin Foundation (grant 2018-J1810250-211305), the Walloon Region through the FRFS-WELBIO strategic research programme (WELBIO-CR-2019C-06), the MSCA-ITN network V.A. Cure no. 814316, the Leducq Foundation Networks of Excellence Program grant "ReVAMP" (LFCR grant 21CVD03), the European Union's Horizon 2020 research and innovation programme under grant agreement no. 874708 (Theralymph), the Swiss National Science Foundation under the Sinergia project no. CRSII5_193694, and a Pierre M. fellowship.
    Keywords:  Angiogenesis; Clinical Trials; Genetic diseases; Lymphomas
    DOI:  https://doi.org/10.1172/jci.insight.173095
  11. Cell Chem Biol. 2023 Oct 26. pii: S2451-9456(23)00367-7. [Epub ahead of print]
    Pharmacologic activation of a compensatory integrated stress response kinase promotes mitochondrial remodeling in PERK-deficient cells.
    Valerie Perea, Kelsey R Baron, Vivian Dolina, Giovanni Aviles, Grace Kim, Jessica D Rosarda, Xiaoyan Guo, Martin Kampmann, R Luke Wiseman.
      The integrated stress response (ISR) comprises the eIF2α kinases PERK, GCN2, HRI, and PKR, which induce translational and transcriptional signaling in response to diverse insults. Deficiencies in PERK signaling lead to mitochondrial dysfunction and contribute to the pathogenesis of numerous diseases. We define the potential for pharmacologic activation of compensatory eIF2α kinases to rescue ISR signaling and promote mitochondrial adaptation in PERK-deficient cells. We show that the HRI activator BtdCPU and GCN2 activator halofuginone promote ISR signaling and rescue ER stress sensitivity in PERK-deficient cells. However, BtdCPU induces mitochondrial depolarization, leading to mitochondrial fragmentation and activation of the OMA1-DELE1-HRI signaling axis. In contrast, halofuginone promotes mitochondrial elongation and adaptive mitochondrial respiration, mimicking regulation induced by PERK. This shows halofuginone can compensate for deficiencies in PERK signaling and promote adaptive mitochondrial remodeling, highlighting the potential for pharmacologic ISR activation to mitigate mitochondrial dysfunction and motivating the pursuit of highly selective ISR activators.
    Keywords:  ISR; UPR; integrated stress response; pharmacologic activator; stress-responsive signaling pathway; unfolded protein response
    DOI:  https://doi.org/10.1016/j.chembiol.2023.10.006
  12. Vascul Pharmacol. 2023 Nov 03. pii: S1537-1891(23)00101-5. [Epub ahead of print]153 107241
    Tissue-specific Cre driver mice to study vascular diseases.
    Qing Rex Lyu, Kailong Fu.
      Vascular diseases, including atherosclerosis and abdominal aneurysms, are the primary cause of mortality and morbidity among the elderly worldwide. The life quality of patients is significantly compromised due to inadequate therapeutic approaches and limited drug targets. To expand our comprehension of vascular diseases, gene knockout (KO) mice, especially conditional knockout (cKO) mice, are widely used for investigating gene function and mechanisms of action. The Cre-loxP system is the most common method for generating cKO mice. Numerous Cre driver mice have been established to study the main cell types that compose blood vessels, including endothelial cells, smooth muscle cells, and fibroblasts. Here, we first discuss the characteristics of each layer of the arterial wall. Next, we provide an overview of the representative Cre driver mice utilized for each of the major cell types in the vessel wall and their most recent applications in vascular biology. We then go over Cre toxicity and discuss the practical methods for minimizing Cre interference in experimental outcomes. Finally, we look into the future of tissue-specific Cre drivers by introducing the revolutionary single-cell RNA sequencing and dual recombinase system.
    Keywords:  Conditional knockout mice; Cre toxicity; Cre-loxP system; Endothelial cells; Smooth muscle cells; Vascular disease
    DOI:  https://doi.org/10.1016/j.vph.2023.107241
  13. Stem Cells. 2023 Nov 01. pii: sxad079. [Epub ahead of print]
    Selective Inhibition of mTORC1 Signaling Supports the Development and Maintenance of Pluripotency.
    Jin Koo Kim, Luis G Villa-Diaz, Thomas L Saunders, Ruiz P Saul, Suraj Timilsina, Fei Liu, Yuji Mishina, Paul H Krebsbach.
      Insight into the molecular mechanisms governing the development and maintenance of pluripotency is important for understanding early development and the use of stem cells in regenerative medicine. We demonstrate the selective inhibition of mTORC1 signaling is important for developing the inner cell mass (ICM) and the self-renewal of human embryonic stem cells. S6K suppressed the expression and function of pluripotency-related transcription factors (PTFs) OCT4, SOX2, and KLF4 through phosphorylation and ubiquitin proteasome-mediated protein degradation, indicating that S6K inhibition is required for pluripotency. PTFs inhibited mTOR signaling. The phosphorylation of S6 was decreased in PTF-positive cells of the ICM in embryos. Activation of mTORC1 signaling blocked ICM formation and the selective inhibition of S6K by rapamycin increased the ICM size in mouse blastocysts. Thus, selective inhibition of mTORC1 signaling supports the development and maintenance of pluripotency.
    Keywords:  4E-BP1; Human pluripotent stem cells (hPSCs); S6; S6K; inner cell mass (ICM); pluripotency-related transcription factors (PTFs)
    DOI:  https://doi.org/10.1093/stmcls/sxad079
  14. Nat Cell Biol. 2023 Nov;25(11): 1557-1560
    Towards inclusive and sustainable scientific meetings.
    Silke Blair Chalmers, Suzanne Madgwick, Bethan Lloyd-Lewis, Urszula Lucja McClurg, Salah Elias, Peter Andersen, Ralitsa Madsen, Vaishnavi Ananthanarayanan, Felicity Mae Davis.
      
    DOI:  https://doi.org/10.1038/s41556-023-01222-9
  15. Front Cell Dev Biol. 2023 ;11 1321316
    Editorial: Single cell dynamics and cell cycle length variation.
    Dirk Loeffler.
      
    Keywords:  cell cycle; cell division; cell fate decisions; cilia; hematopoietic stem cells; pluripotent stem cells; single-cell dynamics; transcription factor condensates
    DOI:  https://doi.org/10.3389/fcell.2023.1321316
  16. Cell. 2023 Nov 01. pii: S0092-8674(23)01128-5. [Epub ahead of print]
    RNA polymerase II dynamics and mRNA stability feedback scale mRNA amounts with cell size.
    Matthew P Swaffer, Georgi K Marinov, Huan Zheng, Lucas Fuentes Valenzuela, Crystal Yee Tsui, Andrew W Jones, Jessica Greenwood, Anshul Kundaje, William J Greenleaf, Rodrigo Reyes-Lamothe, Jan M Skotheim.
      A fundamental feature of cellular growth is that total protein and RNA amounts increase with cell size to keep concentrations approximately constant. A key component of this is that global transcription rates increase in larger cells. Here, we identify RNA polymerase II (RNAPII) as the limiting factor scaling mRNA transcription with cell size in budding yeast, as transcription is highly sensitive to the dosage of RNAPII but not to other components of the transcriptional machinery. Our experiments support a dynamic equilibrium model where global RNAPII transcription at a given size is set by the mass action recruitment kinetics of unengaged nucleoplasmic RNAPII to the genome. However, this only drives a sub-linear increase in transcription with size, which is then partially compensated for by a decrease in mRNA decay rates as cells enlarge. Thus, limiting RNAPII and feedback on mRNA stability work in concert to scale mRNA amounts with cell size.
    Keywords:  RNA; RNA biology; RNA polymerase II; cell biology; cell size; size scaling; systems biology; transcription
    DOI:  https://doi.org/10.1016/j.cell.2023.10.012
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