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Neurogenic Decisions Require a Cell Cycle Independent Function of The
RESEARCH ARTICLE Neurogenic decisions require a cell cycle independent function of the CDC25B phosphatase Fre´ de´ ric Bonnet1†, Angie Molina1†, Me´ lanie Roussat1, Manon Azais2, Sophie Bel-Vialar1, Jacques Gautrais2, Fabienne Pituello1*, Eric Agius1* 1Centre de Biologie du De´veloppement, Centre de Biologie Inte´grative, Universite´ de Toulouse, CNRS, UPS, Toulouse, France; 2Centre de Recherches sur la Cognition Animale, Centre de Biologie Inte´grative., Universite´ de Toulouse, CNRS, UPS, Toulouse, France Abstract A fundamental issue in developmental biology and in organ homeostasis is understanding the molecular mechanisms governing the balance between stem cell maintenance and differentiation into a specific lineage. Accumulating data suggest that cell cycle dynamics play a major role in the regulation of this balance. Here we show that the G2/M cell cycle regulator CDC25B phosphatase is required in mammals to finely tune neuronal production in the neural tube. We show that in chick neural progenitors, CDC25B activity favors fast nuclei departure from the apical surface in early G1, stimulates neurogenic divisions and promotes neuronal differentiation. We design a mathematical model showing that within a limited period of time, cell cycle length modifications cannot account for changes in the ratio of the mode of division. Using a CDC25B point mutation that cannot interact with CDK, we show that part of CDC25B activity is independent *For correspondence: of its action on the cell cycle. [email protected] (FP); [email protected] (EA) †These authors contributed equally to this work Introduction In multicellular organisms, managing the development, homeostasis and regeneration of tissues Competing interests: The requires the tight control of self-renewal and differentiation of stem/progenitor cells. -
Investigating the Role of Cdk11in Animal Cytokinesis
Investigating the Role of CDK11 in Animal Cytokinesis by Thomas Clifford Panagiotou A thesis submitted in conformity with the requirements for the degree of Master of Science Department of Molecular Genetics University of Toronto © Copyright by Thomas Clifford Panagiotou (2020) Investigating the Role of CDK11 in Animal Cytokinesis Thomas Clifford Panagiotou Master of Science Department of Molecular Genetics University of Toronto 2020 Abstract Finely tuned spatio-temporal regulation of cell division is required for genome stability. Cytokinesis constitutes the final stages of cell division, from chromosome segregation to the physical separation of cells, abscission. Abscission is tightly regulated to ensure it occurs after earlier cytokinetic events, like the maturation of the stem body, the regulatory platform for abscission. Active Aurora B kinase enforces the abscission checkpoint, which blocks abscission until chromosomes have been cleared from the cytokinetic machinery. Currently, it is unclear how this checkpoint is overcome. Here, I demonstrate that the cyclin-dependent kinase CDK11 is required for cytokinesis. Both inhibition and depletion of CDK11 block abscission. Furthermore, the mitosis-specific CDK11p58 kinase localizes to the stem body, where its kinase activity rescues the defects of CDK11 depletion and inhibition. These results suggest a model whereby CDK11p58 antagonizes Aurora B kinase to overcome the abscission checkpoint to allow for successful completion of cytokinesis. ii Acknowledgments I am very grateful for the support of my family and friends throughout my studies. I would also like to express my deep gratitude to Wilde Lab members, both past and present, for their advice and collaboration. In particular, I am very grateful to Matthew Renshaw, whose work comprises part of this thesis. -
Mitosis Vs. Meiosis
Mitosis vs. Meiosis In order for organisms to continue growing and/or replace cells that are dead or beyond repair, cells must replicate, or make identical copies of themselves. In order to do this and maintain the proper number of chromosomes, the cells of eukaryotes must undergo mitosis to divide up their DNA. The dividing of the DNA ensures that both the “old” cell (parent cell) and the “new” cells (daughter cells) have the same genetic makeup and both will be diploid, or containing the same number of chromosomes as the parent cell. For reproduction of an organism to occur, the original parent cell will undergo Meiosis to create 4 new daughter cells with a slightly different genetic makeup in order to ensure genetic diversity when fertilization occurs. The four daughter cells will be haploid, or containing half the number of chromosomes as the parent cell. The difference between the two processes is that mitosis occurs in non-reproductive cells, or somatic cells, and meiosis occurs in the cells that participate in sexual reproduction, or germ cells. The Somatic Cell Cycle (Mitosis) The somatic cell cycle consists of 3 phases: interphase, m phase, and cytokinesis. 1. Interphase: Interphase is considered the non-dividing phase of the cell cycle. It is not a part of the actual process of mitosis, but it readies the cell for mitosis. It is made up of 3 sub-phases: • G1 Phase: In G1, the cell is growing. In most organisms, the majority of the cell’s life span is spent in G1. • S Phase: In each human somatic cell, there are 23 pairs of chromosomes; one chromosome comes from the mother and one comes from the father. -
On Cellular Automaton Approaches to Modeling Biological Cells
ON CELLULAR AUTOMATON APPROACHES TO MODELING BIOLOGICAL CELLS MARK S. ALBER∗, MARIA A. KISKOWSKIy , JAMES A. GLAZIERz , AND YI JIANGx Abstract. We discuss two different types of Cellular Automata (CA): lattice-gas- based cellular automata (LGCA) and the cellular Potts model (CPM), and describe their applications in biological modeling. LGCA were originally developed for modeling ideal gases and fluids. We describe several extensions of the classical LGCA model to self-driven biological cells. In partic- ular, we review recent models for rippling in myxobacteria, cell aggregation, swarming, and limb bud formation. These LGCA-based models show the versatility of CA in modeling and their utility in addressing basic biological questions. The CPM is a more sophisticated CA, which describes individual cells as extended objects of variable shape. We review various extensions to the original Potts model and describe their application to morphogenesis; the development of a complex spatial struc- ture by a collection of cells. We focus on three phenomena: cell sorting in aggregates of embryonic chicken cells, morphological development of the slime mold Dictyostelium discoideum and avascular tumor growth. These models include intercellular and extra- cellular interactions, as well as cell growth and death. 1. Introduction. Cellular automata (CA) consist of discrete agents or particles, which occupy some or all sites of a regular lattice. These par- ticles have one or more internal state variables (which may be discrete or continuous) and a set of rules describing the evolution of their state and position (in older models, particles usually occupied all lattice sites, one particle per node, and did not move). -
Epithelial Cell Death Analysis of Cell Cycle by Flow Cytometry White Paper
Epithelial Cell Death Analysis of Cell Cycle by Flow Cytometry White Paper Authors: Savithri Balasubramanian, John Tigges, Vasilis Toxavidis, Heidi Mariani. Affiliation: Beth Israel Deaconess Medical Center Harvard Stem Cell Institute a Beckman Coulter Life Sciences: Epithelial Cell Death Analysis of Cell Cycle by Flow Cytometry PRINCIPAL OF TECHNIQUE Background: Cell cycle, or cell-division cycle, is the series of events that takes place in a cell leading to its division and duplication (replication). In cells without a nucleus (prokaryotic), cell cycle occurs via a process termed binary fission. In cells with a nucleus (eukaryotes), cell cycle can be divided in two brief periods: interphase—during which the cell grows, accumulating nutrients needed for mitosis and duplicating its DNA—and the mitosis (M) phase, during which the cell splits itself into two distinct cells, often called «daughter cells». Cell-division cycle is a vital process by which a single-celled fertilized egg develops into a mature organism, as well as the process by which hair, skin, blood cells, and some internal organs are renewed. Cell cycle consists of four distinct phases: G1 phase, S phase (synthesis), G2 phase (collectively known as interphase) and M phase (mitosis). M phase is itself composed of two tightly coupled processes: mitosis, in which the cell’s chromosomes are divided between the two daughter cells, and cytokinesis, in which the cell’s cytoplasm divides in half forming distinct cells. Activation of each phase is dependent on the proper progression and completion of the previous one. Cells that have temporarily or reversibly stopped dividing are said to have entered a state of quiescence called G0 phase. -
Proteomic and Bioinformatic Investigation of Altered Pathways in Neuroglobin-Deficient Breast Cancer Cells
molecules Article Proteomic and Bioinformatic Investigation of Altered Pathways in Neuroglobin-Deficient Breast Cancer Cells Michele Costanzo 1,2 , Marco Fiocchetti 3 , Paolo Ascenzi 3, Maria Marino 3 , Marianna Caterino 1,2,* and Margherita Ruoppolo 1,2,* 1 Department of Molecular Medicine and Medical Biotechnology, School of Medicine, University of Naples Federico II, 80131 Naples, Italy; [email protected] 2 CEINGE—Biotecnologie Avanzate S.C.Ar.L., 80145 Naples, Italy 3 Department of Science, University Roma Tre, 00146 Rome, Italy; marco.fi[email protected] (M.F.); [email protected] (P.A.); [email protected] (M.M.) * Correspondence: [email protected] (M.C.); [email protected] (M.R.) Abstract: Neuroglobin (NGB) is a myoglobin-like monomeric globin that is involved in several processes, displaying a pivotal redox-dependent protective role in neuronal and extra-neuronal cells. NGB remarkably exerts its function upon upregulation by NGB inducers, such as 17β-estradiol (E2) and H2O2. However, the molecular bases of NGB’s functions remain undefined, mainly in non- neuronal cancer cells. Human MCF-7 breast cancer cells with a knocked-out (KO) NGB gene obtained using CRISPR/Cas9 technology were analyzed using shotgun label-free quantitative proteomics in comparison with control cells. The differential proteomics experiments were also performed after treatment with E2, H2O2, and E2 + H2O2. All the runs acquired using liquid chromatography–tandem mass spectrometry were elaborated within the same MaxQuant analysis, leading to the quantification Citation: Costanzo, M.; Fiocchetti, of 1872 proteins in the global proteomic dataset. Then, a differentially regulated protein dataset was M.; Ascenzi, P.; Marino, M.; Caterino, M.; Ruoppolo, M. -
Role of Cyclin-Dependent Kinase 1 in Translational Regulation in the M-Phase
cells Review Role of Cyclin-Dependent Kinase 1 in Translational Regulation in the M-Phase Jaroslav Kalous *, Denisa Jansová and Andrej Šušor Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Rumburska 89, 27721 Libechov, Czech Republic; [email protected] (D.J.); [email protected] (A.Š.) * Correspondence: [email protected] Received: 28 April 2020; Accepted: 24 June 2020; Published: 27 June 2020 Abstract: Cyclin dependent kinase 1 (CDK1) has been primarily identified as a key cell cycle regulator in both mitosis and meiosis. Recently, an extramitotic function of CDK1 emerged when evidence was found that CDK1 is involved in many cellular events that are essential for cell proliferation and survival. In this review we summarize the involvement of CDK1 in the initiation and elongation steps of protein synthesis in the cell. During its activation, CDK1 influences the initiation of protein synthesis, promotes the activity of specific translational initiation factors and affects the functioning of a subset of elongation factors. Our review provides insights into gene expression regulation during the transcriptionally silent M-phase and describes quantitative and qualitative translational changes based on the extramitotic role of the cell cycle master regulator CDK1 to optimize temporal synthesis of proteins to sustain the division-related processes: mitosis and cytokinesis. Keywords: CDK1; 4E-BP1; mTOR; mRNA; translation; M-phase 1. Introduction 1.1. Cyclin Dependent Kinase 1 (CDK1) Is a Subunit of the M Phase-Promoting Factor (MPF) CDK1, a serine/threonine kinase, is a catalytic subunit of the M phase-promoting factor (MPF) complex which is essential for cell cycle control during the G1-S and G2-M phase transitions of eukaryotic cells. -
Interaction of Cdc2 and Rum1 Regulates Start and S-Phase in Fission Yeast
Journal of Cell Science 108, 3285-3294 (1995) 3285 Printed in Great Britain © The Company of Biologists Limited 1995 JCS8905 Interaction of cdc2 and rum1 regulates Start and S-phase in fission yeast Karim Labib1,2,3, Sergio Moreno3 and Paul Nurse1,2,* 1ICRF Cell Cycle Laboratory, Department of Biochemistry, University of Oxford, Oxford, OX1 1QU, UK 2ICRF, PO Box 123, Lincolns Inn Fields, London WC2A 3PX, UK 3Instituto de Microbiologia-Bioquimica, CSIC/Universidad de Salamanca, Edificio Departamental, Campus Miguel de Unamuno, 37007 Salamanca, Spain *Author for correspondence SUMMARY The p34cdc2 kinase is essential for progression past Start in rum1, can disrupt the dependency of S-phase upon mitosis, the G1 phase of the fission yeast cell cycle, and also acts in resulting in an extra round of S-phase in the absence of G2 to promote mitotic entry. Whilst very little is known mitosis. We show that cdc2 and rum1 interact in this about the G1 function of cdc2, the rum1 gene has recently process, and describe dominant cdc2 mutants causing been shown to encode an important regulator of Start in multiple rounds of S-phase in the absence of mitosis. We fission yeast, and a model for rum1 function suggests that suggest that interaction of rum1 and cdc2 regulates Start, it inhibits p34cdc2 activity. Here we present genetic data and this interaction is important for the regulation of S- cdc2 suggesting that rum1 maintains p34 in a pre-Start G1 phase within the cell cycle. form, inhibiting its activity until the cell achieves the critical mass required for Start, and find that in the absence of rum1 p34cdc2 has increased Start activity in vivo. -
Recruitment of Leukemic Cells from G 0 Phase of the Cell Cycle By
Leukemia (2003) 17, 2049–2059 & 2003 Nature Publishing Group All rights reserved 0887-6924/03 $25.00 www.nature.com/leu CORRESPONDENCE Recruitment of leukemic cells from G0 phase of the cell cycle by interferons results in conversion of resistance to daunorubicin Leukemia (2003) 17, 2049–2051. doi:10.1038/sj.leu.2403085 were not affected by daunorubicin-induced cell death. To analyze whether a similar cell cycle-specific sensitivity to Ara-C TO THE EDITOR and daunorubicin was observed in other leukemic cell lines, we analyzed four acute lymphoblastic leukemia cell lines and one Although the majority of patients with acute myeloid leukemia CML blast crisis cell line, which were generated in our (AML) responds to initial treatment, relapse of the disease occurs laboratory by culturing of leukemic blasts from five different in a significant percentage of these patients.1 The cell cycle patients in serum-free medium at high cell concentrations until status of leukemic cells may play an important role in the spontaneous sustained proliferation of the leukemic cells response to treatment of leukemic cells. Especially, the broadly occurred. These cell lines cytogenetically and phenotypically used cytotoxic agent Cytarabine (Ara-C) has been demonstrated resembled the primary leukemia. Figure 1b shows the median to exert its action via intercalation into the DNA of cells cell cycle distribution within these cell lines after incubation for À6 specifically in the S phase of the cell cycle, and sensitivity to this 48 h in medium alone, or in medium containing 1 Â 10 M Ara- agent is therefore described to be specific for cells in active cell C or daunorubicin. -
The Cell Cycle & Mitosis
The Cell Cycle & Mitosis Cell Growth The Cell Cycle is G1 phase ___________________________________ _______________________________ During the Cell Cycle, a cell ___________________________________ ___________________________________ Anaphase Cell Division ___________________________________ Mitosis M phase M ___________________________________ S phase replication DNA Interphase Interphase is ___________________________ ___________________________________ G2 phase Interphase is divided into three phases: ___, ___, & ___ G1 Phase S Phase G2 Phase The G1 phase is a period of The S phase replicates During the G2 phase, many of activity in which cells _______ ________________and the organelles and molecules ____________________ synthesizes _______ molecules. required for ____________ __________ Cells will When DNA replication is ___________________ _______________ and completed, _____________ When G2 is completed, the cell is synthesize new ___________ ____________________ ready to enter the ____________________ ____________________ ____________________ ____________________ ____________________ Mitosis are divided into four phases: _____________, ______________, _____________, & _____________ Below are cells in two different phases of the cell cycle, fill in the blanks using the word bank: Chromatin Nuclear Envelope Chromosome Sister Chromatids Nucleolus Spinder Fiber Centrosome Centrioles 5.._________ 1.__________ v 6..__________ 2.__________ 7.__________ 3.__________ 8..__________ 4.__________ v The Cell Cycle & Mitosis Microscope Lab: -
P53 Phosphorylation and Association with Murine Double Minute 2, C-Jun
[CANCER RESEARCH 60, 896–900, February 15, 2000] Advances in Brief p53 Phosphorylation and Association with Murine Double Minute 2, c-Jun ARF NH2-Terminal Kinase, p14 , and p300/CBP during the Cell Cycle and after Exposure to Ultraviolet Irradiation1 Thomas Buschmann, Victor Adler, Ekaterina Matusevich, Serge Y. Fuchs, and Ze’ev Ronai2 Ruttenberg Cancer Center, Mount Sinai School of Medicine, New York, New York 10029 Abstract though the mechanisms underlying the ability of p53 to elicit such opposing effects are yet to be identified, independent studies point to p53 phosphorylation and association with proteins is implicated in its a different set of p53 regulators and effectors that are affected by p53 stability and activity. We have compared the association of DNA-bound in each of these scenarios. and overall pools of p53 with murine double minute 2 (Mdm2), c-Jun NH -terminal kinase (JNK), p300/CBP, and p14ARF during cell cycle In studying the regulation of p53 stability, we demonstrated previ- 2 ously that, in Swiss 3T3 cells, JNK and Mdm2 target p53 degradation progression. Whereas DNA-bound p53 associates with JNK at G0-G1 and in different phases of the cell cycle (7). In this study, we have with Mdm2 and p300 during S and G2-M phases, the general pool of p53 was found in complex with JNK and Mdm2 almost throughout the cell compared the association of subpopulations of p53 with proteins cycle. Phosphorylation of p53 at serines 9, 15, and 20 is at the highest levels implicated in stability and activity of p53 and monitored the pattern of at G1 and at serines 37 and 392 during G2-M phase. -
Label the Parts of the Cell Cycle Diagram and Briefly Describe What Is Happening: a B C D E F G H I J 1) Chromosomes Move To
Label the parts of the cell cycle diagram and briefly describe what is happening: A Interphase - cell is growing and preparing for division. B G1 - growth and normal cell function C S - DNA replication D G2 - growth, preparation for division, duplicate organelles E Prophase - nuclear envelope dissolves, mitotic apparatus set up, DNA condenses. F Metaphase - chromosome line up in center G Anaphase - sister chromatids separate and are pulled to poles of cell. H Telophase - nuclei reform, DNA relaxes into chromatin, cleaveage furrow or cell plate forms I Mitosis - nuclear division J Cytokinesis - the rest of the cell divides 1) Chromosomes move to the middle of the cell during what phase? 2) What are sister chromatids? 3) What holds the chromatids together? 4) When do the sister chromatids separate? 5) During which phase do chromosomes first become visible? 6) During which phase does the cleavage furrow start forming? 7) What is another name for mitosis? 8) What is the structure that breaks the spindle fiber into 2? 9) What makes up the mitotic apparatus? 10) Complete the table by checking the correct column for each statement. Statement Interphase Mitosis Cell growth occurs Nuclear division occurs Chromosomes are finishing moving into separate daughter cells. Normal functions occur Chromosomes are duplicated DNA synthesis occurs Cytoplasm divides immediately after this period Mitochondria and other organelles are made. The Animal Cell Cycle – Phases are out of order 11) Which cell is in metaphase? 12) Cells A and F show an early and late stage of the same phase of mitosis. What phase is it? 13) In cell A, what is the structure labeled X? 14) In cell F, what is the structure labeled Y? 15) Which cell is not in a phase of mitosis? 16) A new membrane is forming in B.