DOCSLIB.ORG

Influence of ATM Function on Telomere Metabolism

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Influence of ATM Function on Telomere Metabolism Oncogene (1997) 15, 2659 ± 2665 1997 Stockton Press All rights reserved 0950 ± 9232/97 $12.00 In¯uence of ATM function on telomere metabolism Lubomir B Smilenov1, Susan E Morgan2, Wilfredo Mellado1, Satin G Sawant1, Michael B Kastan2 and Tej K Pandita1 1Center for Radiological Research, College of Physicians and Surgeons, Columbia University, New York, NY 10032; 2The Johns Hopkins Oncology Center, Baltimore, Maryland 21205, USA The ATM gene product, which is defective in the cancer- associations (TA) also called chromosome end associa- prone disorder ataxia telangiectasia, has been implicated tions, involve telomeres which are essential for the in mitogenic signal transduction, chromosome condensa- stability and complete replication of eukaryotic tion, meiotic recombination and cell cycle control. The chromosomes (Muller, 1938; McClintock, 1941). ATM gene has homology with the TEL1 gene of yeast, Telomeric associations have been linked to genomic mutations of which lead to shortened telomeres. To test instability and carcinogenicity (Pathak et al., 1988; the hypothesis that the ATM gene product is involved in Counter et al., 1992; Pandita et al., 1995, 1996). In telomere metabolism, we examined telomeric associa- contrast to the unstable ends generated by chromo- tions (TA), telomere length, and telomerase activity in some breakage, telomeres cap chromosomes, prevent human cells expressing either dominant-negative or degradation and end-to-end ligation. Mammalian complementing fragments of the ATM gene. The telomeres are composed of tandem arrays of phenotype of RKO colorectal tumor cells expressing (TTAGGG)n repeat sequences and are maintained by ATM fragments containing a leucine zipper (LZ) motif telomerase. Telomeres shorten as a function of age in mimics that of ataxia telangiectasia (A-T) cells. These cells derived from normal blood, skin and colonic transfected RKO cells relative to transfected controls mucosa (Hastie et al., 1990; Lindsey et al., 1991; had a higher frequency of cells with TA and shortened Allsopp et al., 1992;). As a result of this shortening, it telomeres, but no detectable change in telomerase is thought that critical genes at the ends of activity. In addition, the percentage of cells with TA chromosomes either become deleted or are activated, after gamma irradiation was higher in the transfected thus leading to cell death (Levy et al., 1992; Olovnick, RKO cells with dominant negative activity of the ATM 1992). gene, compared to control cells. SV40 transformed Recently, the gene that mediates the disease ataxia ®broblasts derived from an A-T patient and transfected telangiectasia has been cloned and has been designated with a complementing carboxyl terminal kinase region of ATM (AT, mutated) (Savitsky et al., 1995a). The the ATM gene had a reduced frequency of cells with TA, predicted product of the ATM gene open reading with no eect on the telomere length or telomerase frame is a large protein of 3056 amino acids (Savitsky activity. The present studies using isogenic cells with et al., 1995b). The protein shows similarity to several manipulated ATM function demonstrate a role for the yeast and mammalian proteins involved in meiotic ATM gene product in telomere metabolism. recombination and cell cycle progression, namely the products of MEC1 in budding yeast and rad3 in ®ssion Keywords: telomeric associations; telomere length; yeast, and the TOR proteins of yeast and mammals telomerase activity; A-T; LZ; PI-3 kinase domain (Savitsky et al., 1995b; Brush et al., 1996; Morrow et al., 1996). The ATM gene product shares the PI-3 kinase signature of a growing family of proteins involved in the control of cell cycle progression, Introduction processing of DNA damage and maintenance of genomic stability (Greenwell et al., 1995; Keith and Ataxia telangiectasia (A-T) is an autosomal disorder Schreiber, 1995; Hawley and Friend, 1996). The characterized by progressive neurological degeneration, presence of leucine zipper motif in the ATM protein premature aging, growth retardation, speci®c immuno- suggests possible dimerization of the protein or de®ciencies, telangiectasias, high sensitivity to ionizing interaction with additional proteins. Because of ATM radiation, genomic instability and cancer progression homology to TEL1 mutants of yeast (Greenwell et al., (Bridges and Harnden, 1982; Sedgwick and Border, 1995), it has been suggested that mutations in ATM 1991; Harnden, 1994). Cells derived from A-T patients gene could lead to defective telomere maintenance. To exhibit a variety of abnormalities in culture such as test the hypothesis that the ATM gene product hypersensitivity to ionizing radiation, higher require- in¯uences telomere metabolism, the cell types listed in ments for serum growth factors and cytoskeletal Table 1 with dominant-negative, as well as comple- defects (Gatti et al., 1991; Meyn, 1995; Shiloh, 1995). menting activity for ATM function were utilized A-T patients have a high frequency of cells with (Morgan et al., 1997). Expression of the dominant- telomeric associations seen at metaphase and show negative ATM fragments in RKO cells leads to accelerated shortening of telomeres (Kojis et al., 1991; decreased clonogenic survival, increased chromosomal Pandita et al., 1995; Metcalf et al., 1996). Telomeric aberrations and radioresistant DNA synthesis after treatment with ionizing radiation (Morgan et al., 1997). The ATM fragment containing the kinase domain Correspondence: TK Pandita complemented radiosensitivity, the S-phase checkpoint, Received 10 June 1997; revised 23 July 1997; accepted 24 July 1997 irradiation-induced activation of c-Abl and reduced ATM gene and telomeres LB Smilenov et al 2660 Table 1 Description of cells with dominant negative or complementation activity of ATM gene Cell line Cell type Functional activity RKO Colorectal carcinoma cells RKOpBABEpuro RKO with vector only RKOFB2F3 RKO with LZ motif of ATM gene Dominant negative RKOFB2F7 RKO with LZ motif of ATM gene Dominant negative RKOFB2F12 RKO with LZ motif of ATM gene Dominant negative RKOENA/FB2F18 RKO with LZ motif with entire 5' coding region of ATM gene Dominant negative GM5849 SV40 immortalized A-T cells GM5849LXSNeo GM5849 with vector only GM5849P13K4 GM5849 with P13K domain of ATM gene Complementation GM5849P13K10 GM5849 with P13K domain of ATM gene Complementation chromosomal aberrations after treatment with gamma dominant-negative ATM fragment (Table 2). In order rays in SV40 transformed ®broblasts derived from an to determine whether loss of telomeric signals was due A-T patient (Baskaran et al., 1997; Morgan et al., to a reduction in telomere length, we next evaluated 1997). telomere length in these RKO cells at early passage as well as after several population doublings. The telomere length of the RKO cells at the time of Results and Discussion infection was 7.8 kb. The earliest passage transfected cells used for the determination of telomere length were Recently, telomeric associations have been reported as at about 31 population doublings. Telomere length was a consequence of mutations in the UbcD1 gene in determined by measuring the terminal restriction Drosophila melanogaster (Cenci et al., 1997), as well as fragment (TRF) in RsaI and HinfI digested DNA mutations in a telomerase template in Tetrahymena (Figure 3). Three out of the four RKO clones thermophila (Kirk et al., 1997). To determine whether (RKOFB2F3, RKOFB2F7 and RKOENA/FB2F18) ATM function in¯uences TA, we examined metaphase expressing the dominant-negative fragment had statis- spreads and found that RKO cells expressing tically signi®cant shorter mean TRF (Table 2). The dominant-negative fragments exhibited a 3 ± 6-fold rate of telomere shortening varied in the dierent cell higher frequency of cells with TA compared to lines from 80 ± 130 base pairs per population doubling. parental and control RKO cells (Figure 1a). This The one transfected clone (RKOFB2F12) which did abnormal telomeric association phenotype was mani- not have shorter telomeres was the same clone that had fested in both G1- and G2-phase cells as determined by an unaltered telomere signal. To determine the using the premature chromosome condensation techni- in¯uence of the pBABEpuro vector on telomere que (Figure 1a) (Pandita et al., 1995). Since gamma length, we determined the mean TRF of four clones irradiation triggers TA in A-T cells (Pandita and of RKOpBABEpuro. We found that there were no Hittelman, 1993), we also evaluated TA in these cells signi®cant dierences in mean TRF from the RKO after radiation treatment and found that the frequency control among dierent clones of RKOpBABEpuro. of cells with TA also increased in the RKO cells with This suggests that accelerated telomere shortening was the dominant-negative ATM fragments, compared to not due to the presence of the vector. Furthermore, control cells (Figure 1b). when TRF was evaluated with increased population Telomeric association frequencies were also evalu- doublings in the RKO cells expressing the dominant- ated in A-T cells expressing the complementing kinase negative fragments (RKOFB2F3), both a decrease in domain fragment. The kinase-complemented A-T cells telomere signal (data not shown) and a shortening of had a 2 ± 4-fold lower frequency of cells with TA than telomere length (loss of about 22 base pairs per the parental or control-transfected cells in metaphase, population doubling) was observed with the increase as well as in G1 and G2 (Figure 2a). Similarly, in population doubling numbers
Load more

Recommended publications
  • Snapshot: Inositol Phosphates Ace J
    SnapShot: Inositol Phosphates Ace J. Hatch and John D. York HHMI, Pharmacology and Cancer Biology, Biochemistry, Duke University, Durham, NC 27710, USA PLC-dependent IP code GPCR RTK O O O O O O 5-PP-IP4 IP4 5-IP7 O O O O O O PIP2 O IP6K O IP6K O VIP1 O O O 2 O ITPK1 O 13 O PLC 2 O O O O O O O O 4 6 13 O 5 IP3 IPMK IP4 IPMK IP5 IPK1 IP6 1,5-IP8 4 6 O O 5 O O O O O O O O ENZYMES O O O O O O YEAST MAMMALIAN IP3K VIP1 IP6K IPMK PLC1 PLCβ, γ, δ, ε, ζ, η O - IP3KA, B, C - ITPK1 (IP56K) O O O O O O O O IPK2(ARG82) IPMK (IPK2) IP4 IP3 IP4 1-IP7 IPK1 IPK1 (IP5K) INPP5 ITPK1 KCS1 IP6K1, 2, 3 O O O O O O VIP1 VIP1, 2 (PPIP5K1, 2) O O Ion channels Phosphate sensing Transcription Cl- Abundant phosphate MCM1 ARG80 CIC3 P PLASMA MEMBRANE - Pho80 Cl channel Pho4 Kinase Kinase Assembly Pho85 independent CYTOPLASM activity 2 O PIP2 Pho81 13 CYTOPLASM NUCLEUS IPK2 ARG81 4 6 Phosphate starvation MCM1-ArgR O 5 O complex O O IP4 O O O O O O O 1-IP7 Kinase Activation dependent IP3 O O Transcription O O O activated Pho80 IP4 O X Pho4 O O Pho85 Kinase activity IP receptor blocked O 3 ENDOPLASMIC Pho81 RETICULUM Ca2+ CYTOPLASM NUCLEUS NUCLEUS mRNA export and translation Insulin secretion and AKT Embryonic development Translation termination Effects of IP kinase deficiency O IPMK (IPK2): Multiple defects, death by embryonic day 10 (mice) O O Insulin IPK1: Cillia are shortened and immotile IP6 AKT resistance causing patterning defects (zebrash) O O Multiple defects, death by Ribosome O embryonic day 8.5 (mice) GleI eRF1 Insulin GSK3β Dbp5 ITPK1 (IP56K): Neural tube
    [Show full text]
  • Ceramide Kinase Is Upregulated in Metastatic Breast Cancer Cells and Contributes to Migration and Invasion by Activation of PI 3-Kinase and Akt
    International Journal of Molecular Sciences Article Ceramide Kinase Is Upregulated in Metastatic Breast Cancer Cells and Contributes to Migration and Invasion by Activation of PI 3-Kinase and Akt 1,2, 1, 2 2 Stephanie Schwalm y, Martin Erhardt y, Isolde Römer , Josef Pfeilschifter , Uwe Zangemeister-Wittke 1,* and Andrea Huwiler 1,* 1 Institute of Pharmacology, University of Bern, Inselspital, INO-F, CH-3010 Bern, Switzerland; [email protected] (S.S.); [email protected] (M.E.) 2 Institute of General Pharmacology and Toxicology, University Hospital Frankfurt am Main, Goethe-University, Theodor-Stern Kai 7, D-60590 Frankfurt am Main, Germany; [email protected] (I.R.); [email protected] (J.P.) * Correspondence: [email protected] (U.Z.-W.); [email protected] (A.H.); Tel.: +41-31-6323214 (A.H.) These authors contributed equally to this work. y Received: 29 November 2019; Accepted: 12 February 2020; Published: 19 February 2020 Abstract: Ceramide kinase (CerK) is a lipid kinase that converts the proapoptotic ceramide to ceramide 1-phosphate, which has been proposed to have pro-malignant properties and regulate cell responses such as proliferation, migration, and inflammation. We used the parental human breast cancer cell line MDA-MB-231 and two single cell progenies derived from lung and bone metastasis upon injection of the parental cells into immuno-deficient mice. The lung and the bone metastatic cell lines showed a marked upregulation of CerK mRNA and activity when compared to the parental cell line. The metastatic cells also had increased migratory and invasive activity, which was dose-dependently reduced by the selective CerK inhibitor NVP-231.
    [Show full text]
  • Inhibition of Spinach Phosphoribulokinase by DL-Glyceraldehyde by ANTONI R
    Biochem. J. (1976) 153, 613-619 613 Printed in Great Britain Inhibition of Spinach Phosphoribulokinase by DL-Glyceraldehyde By ANTONI R. SLABAS and DAVID A. WALKER Department ofBotany, University ofSheffield, Sheffield S1O 2TN, U.K. (Received 10 September 1975) Spinach chloroplast phosphoribulokinase is inhibited by DL-glyceraldehyde. The in- hibition is non-competitive with respect to ribulose 5-phosphate (Ki 19mM) and ATP (Ki 20mM). The inhibition is discussed in relation to a previously reported inhibition of CO2 assimilation in intact and envelope-free chloroplasts by DL-glyceraldehyde. It is concluded that the inhibition of phosphoribulokinase is insufficient to account for the inhibition, by DL-glyceraldehyde, of 02 evolution with ribose 5-phosphate as substrate and that a further site of inhibition is also present in this system. DL-Glyceraldehyde inhibits photosynthetic carbon glycylglycine buffer, pH7.4, in a total volume of assiniilation by intact chloroplasts and by the re- 13 ml. The reaction was terminated by the addition constituted chloroplast system (Stokes & Walker, of 2ml of 50 % (w/v) trichloroacetic acid and the pH 1972). The inhibition is most pronounced with intact was adjusted to pH8.0 with KOH. After the addition chloroplasts, a concentration of 1OmM being sufficient of Sml of 1.OM-barium acetate the solution was to suppress 02 evolution completely. It is important centrifuged and the precipitate discarded after because DL-glyceraldehyde is the only known inhibi- washing with Sml of water. Cold ethanol (4vol.) was tor of photosynthesis that is entirely without detect- added to the combined supernatants (total volume able effect on photophosphorylation or photo- 25.4ml); the new precipitate was recovered by synthetic electron transport.
    [Show full text]
  • Swiveling Domain Mechanism in Pyruvate Phosphate Dikinase†,‡ Kap Lim,§ Randy J
    Biochemistry 2007, 46, 14845-14853 14845 Swiveling Domain Mechanism in Pyruvate Phosphate Dikinase†,‡ Kap Lim,§ Randy J. Read,| Celia C. H. Chen,§ Aleksandra Tempczyk,§ Min Wei,⊥ Dongmei Ye,⊥ Chun Wu,⊥ Debra Dunaway-Mariano,⊥ and Osnat Herzberg*,§ Center for AdVanced Research in Biotechnology, UniVersity of Maryland Biotechnology Institute, RockVille, Maryland 20850, Department of Haematology, Cambridge Institute for Medical Research, UniVersity of Cambridge, Cambridge, United Kingdom, and Department of Chemistry, UniVersity of New Mexico, Albuquerque, New Mexico ReceiVed September 10, 2007; ReVised Manuscript ReceiVed October 17, 2007 ABSTRACT: Pyruvate phosphate dikinase (PPDK) catalyzes the reversible conversion of phosphoenolpyruvate (PEP), AMP, and Pi to pyruvate and ATP. The enzyme contains two remotely located reaction centers: the nucleotide partial reaction takes place at the N-terminal domain, and the PEP/pyruvate partial reaction takes place at the C-terminal domain. A central domain, tethered to the N- and C-terminal domains by two closely associated linkers, contains a phosphorylatable histidine residue (His455). The molecular architecture suggests a swiveling domain mechanism that shuttles a phosphoryl group between the two reaction centers. In an early structure of PPDK from Clostridium symbiosum, the His445-containing domain (His domain) was positioned close to the nucleotide binding domain and did not contact the PEP/pyruvate- binding domain. Here, we present the crystal structure of a second conformational state of C. symbiosum PPDK with the His domain adjacent to the PEP-binding domain. The structure was obtained by producing a three-residue mutant protein (R219E/E271R/S262D) that introduces repulsion between the His and nucleotide-binding domains but preserves viable interactions with the PEP/pyruvate-binding domain.
    [Show full text]
  • Ceramides: Nutrient Signals That Drive Hepatosteatosis
    J Lipid Atheroscler. 2020 Jan;9(1):50-65 Journal of https://doi.org/10.12997/jla.2020.9.1.50 Lipid and pISSN 2287-2892·eISSN 2288-2561 Atherosclerosis Review Ceramides: Nutrient Signals that Drive Hepatosteatosis Scott A. Summers Department of Nutrition and Integrative Physiology, University of Utah, Salt Lake City, UT, USA Received: Sep 24, 2019 ABSTRACT Revised: Nov 4, 2019 Accepted: Nov 10, 2019 Ceramides are minor components of the hepatic lipidome that have major effects on liver Correspondence to function. These products of lipid and protein metabolism accumulate when the energy needs Scott A. Summers of the hepatocyte have been met and its storage capacity is full, such that free fatty acids start Department of Nutrition and Integrative to couple to the sphingoid backbone rather than the glycerol moiety that is the scaffold for Physiology, University of Utah, 15N 2030E, Salt Lake City, UT 84112, USA. glycerolipids (e.g., triglycerides) or the carnitine moiety that shunts them into mitochondria. E-mail: [email protected] As ceramides accrue, they initiate actions that protect cells from acute increases in detergent- like fatty acids; for example, they alter cellular substrate preference from glucose to lipids Copyright © 2020 The Korean Society of Lipid and they enhance triglyceride storage. When prolonged, these ceramide actions cause insulin and Atherosclerosis. This is an Open Access article distributed resistance and hepatic steatosis, 2 of the underlying drivers of cardiometabolic diseases. under the terms of the Creative Commons Herein the author discusses the mechanisms linking ceramides to the development of insulin Attribution Non-Commercial License (https:// resistance, hepatosteatosis and resultant cardiometabolic disorders.
    [Show full text]
  • Glucokinase September 15, 2005
    Glucokinase September 15, 2005 Summary Glucokinase phosphorylates glucose into glucose 6-phosphate using ATP as the phosphate group donor, according to the reaction glucose + ATP Æ glucose 6-phosphate + ADP This protocol describes an indirect assay to determine the activity of glucokinase. Glucose 6- phosphate formed by glucokinase is measured by the formation of NADPH in presence of glucose 6-phosphate dehydrogenase. Solutions Required 1. 100 mM tris HCl pH 7.4 Stock solution could be used 2. 100 mM MgSO4·7H2O Stock solution could be used 3. 10 mM glucose Must be prepared fresh 4. 20 mM ATP Must be prepared fresh 5. 10 mM NADP Must be prepared fresh 6. a solution containing 10 U of glucose 6-phosphate dehydrogenase per 50 μL in tris HCl pH 7.4 buffer. This solution is prepared by mixing 40 μL of a 5000 U/mL solution (Sigma G8529) with 960 μL tris buffer. Preparation of Cell Extract Follow general protocol described in Preparation of Cell Extract. The cell extract should be suspended in tris buffer after pelletization. Spectrophotometer Turn on the ultraviolet bulb on the spectrophotometer (Beckman DU50) and wait 10 minutes for warm-up. Select the kinetics-time window on the instrument. Load the method "A:/nadh" ". These methods each have a run-time of 120 s, a temperature of 37°C, a wavelength of 340 nm and use 2 autosamplers. Procedure 1. For each assay, prepare the two cocktails shown in the following table into two separate quartz cuvettes. Keep them on ice. Volume (μL) added to: Solution Control Experimental tris 500 500 DI H2O 200 100 MgSO4 100 100 glucose 0 100 ATP 50 50 NADP 50 50 G6P dehydrogenase 50 50 2.
    [Show full text]
  • Inhibition of Human Telomerase Enhances the Effect of the Tyrosine Kinase Inhibitor, Imatinib, in BCR-ABL-Positive Leukemia Cells1
    Vol. 8, 3341–3347, November 2002 Clinical Cancer Research 3341 Advances in Brief Inhibition of Human Telomerase Enhances the Effect of the Tyrosine Kinase Inhibitor, Imatinib, in BCR-ABL-positive Leukemia Cells1 Tetsuzo Tauchi,2 Akihiro Nakajima, by only two amino acids, exhibited normal morphology. The Goro Sashida, Takashi Shimamoto, evidence of apoptosis in the telomerase-inhibited cells was Junko H. Ohyashiki, Kenji Abe, determined by flow cytometric analysis with APO2.7 mono- clonal antibody. We also observed enhanced induction of Kohtaro Yamamoto, and Kazuma Ohyashiki apoptosis by imatinib seen in DN-hTERT-expressing K562 First Department of Internal Medicine, Tokyo Medical University, cells, as compared with WT-hTERT-expressing cells. Tokyo, 160-0023 [T. T., A. N., G. S., T. S., K. O.]; Intractable Conclusions: These results demonstrate that disruption Disease Research Center, Tokyo Medical University, Shinjuku-ku, Tokyo 160-0023 [J. H. O.]; and Department of Virology, Medical of telomere maintenance limits the cellular life span of leu- Research Institute, Tokyo Medical and Dental University, Tokyo, kemia cells and show that the combined use of imatinib and 113-8519 [K. A., K. Y.], Japan telomere maintenance inhibition may be effective in the treatment of BCR-ABL-positive leukemia. Abstract Purpose: Telomerase is a ribonucleoprotein enzyme Introduction that maintains protective structures at the ends of eukary- BCR-ABL is a chimeric oncoprotein generated by recip- otic chromosomes. Earlier findings have supported an rocal translocation between chromosomes 9 and 22 and impli- association between progressive telomere shortening in cated in the pathogenesis of Ph3-positive leukemia (1). BCR- the chronic phase of chronic myelogenous leukemia and the ABL fusion proteins exhibit elevated tyrosine kinase activity up-regulation of telomerase activity occurring late in and transforming properties (2, 3).
    [Show full text]
  • Standard Operating Procedure Procedure in Vitro Kinase Assay
    Standard Operating Procedure Procedure In Vitro Kinase Assay with [γ-32P] - ATP Department Location SOP Prepared By: Section 1: Purpose Kinase assays are used to detect the activity of specific kinases from cells. Kinases are a group of enzymes that modifies a target substrate by phosphorylation. In this assay, the kinase removes the radioactively labeled phosphate group from [γ-32P] - ATP and adds it to the target substrate. Kinase activity is measured by running an SDS PAGE (gel electrophoresis for proteins) and visualizing the results with an imaging technique which quantifies the amount of radiolabeled substrate. Given kinases’ essential role in cell metabolism, signaling, etc, this assay can be applied virtually to any biological research that wishes to investigate enzymatic activity within a cellular process. Some studies include measuring yeast kinases to understand chromosomal segregation1 and measuring protein kinase R (PKR) activity2, an important modulator of chronic metabolic inflammation associated to obesity, diabetes, and cardiovascular disease. Section 2: Personal Protective Equipment and Survey Equipment PPE: • Lab coat • Nitrile Gloves • Heat resistant gloves • Safety Glasses • Closed-toe shoes Other Equipment: • Geiger counter with pancake probe • Personal chest dosimeter • Personal finger dosimeter Section 3: Radioactive Material ATP, [γ-32P], Supplier: Perkin Elmer Starting activity: 10 uCi/uL Typical use quantities: 100 uCi - 1mCi per experiment • Radioactive material exposure (RAM) time on benchtop: ~30 minutes Activity used per experiment: _______________ RAM handling time: _______________ Frequency of experiment: _______________ Section 4: Potential Hazards • 32P is a high-energy beta emitter and has a half-life of 14.29 days. 32P can present a substantial skin and eye dose hazard.
    [Show full text]
  • Role of Ceramide Kinase in Breast Cancer Progression
    University of Pennsylvania ScholarlyCommons Publicly Accessible Penn Dissertations 2014 Role of Ceramide Kinase in Breast Cancer Progression Ania Warczyk Payne University of Pennsylvania, [email protected] Follow this and additional works at: https://repository.upenn.edu/edissertations Part of the Cell Biology Commons, Molecular Biology Commons, and the Pharmacology Commons Recommended Citation Payne, Ania Warczyk, "Role of Ceramide Kinase in Breast Cancer Progression" (2014). Publicly Accessible Penn Dissertations. 1402. https://repository.upenn.edu/edissertations/1402 This paper is posted at ScholarlyCommons. https://repository.upenn.edu/edissertations/1402 For more information, please contact [email protected]. Role of Ceramide Kinase in Breast Cancer Progression Abstract Recurrent breast cancer is typically an incurable disease and, as such, is disproportionately responsible for deaths from this disease. Recurrent breast cancers arise from the pool of disseminated tumor cells (DTCs) that survive adjuvant or neoadjuvant therapy, and patients with detectable DTCs following therapy are at substantially increased risk for recurrence. Consequently, the identification of pathways that contribute to the survival of breast cancer cells following therapy could aid in the development of more effective therapies that decrease the burden of residual disease and thereby reduce the risk of breast cancer recurrence. We now report that Ceramide Kinase (Cerk) is required for mammary tumor recurrence following HER2/neu pathway inhibition and is spontaneously up-regulated during tumor recurrence in multiple genetically engineered mouse models for breast cancer. We find that Cerk is apidlyr up-regulated in tumor cells following HER2/neu down-regulation or treatment with adriamycin and that Cerk is required for tumor cell survival following HER2/neu down-regulation.
    [Show full text]
  • Pyruvate Orthophosphate Dikinase
    Chapter 15 Structure, Function, and Post-translational Regulation of C4 Pyruvate Orthophosphate Dikinase Chris J. Chastain Department of Biosciences, Minnesota State University-Moorhead , Moorhead , MN 56563 , USA Summary .............................................................................................................................................................. 301 I. Introduction .................................................................................................................................................... 302 A. Role of PPDK in C 4 Plants ...................................................................................................................... 302 B. PPDK Enzyme Properties ...................................................................................................................... 302 1. Catalysis as Related to Structure ..................................................................................................... 302 2. Oligomeric Structure and Tetramer Dissociation at Cool Temperatures ........................................... 304 3. Substrate Km s for C 4 PPDK .............................................................................................................. 304 C. PPDK as a Rate-Limiting Enzyme of the C 4 Pathway ............................................................................ 304 II. Post-translational Regulation of C 4 PPDK ..................................................................................................... 305 A. Light/Dark
    [Show full text]
  • The Serine/Threonine Protein Kinase (Akt)/ Protein Kinase B (Pkb) Signaling Pathway in Breast Cancer
    Journal of Mind and Medical Sciences Volume 7 Issue 1 Article 7 2020 The Serine/Threonine Protein Kinase (Akt)/ Protein Kinase B (PkB) Signaling Pathway in Breast Cancer Daniela Miricescu FACULTY OF DENTAL MEDICINE, DEPARTMENT OF BIOCHEMISTRY, BUCHAREST, ROMANIA Camelia Cristina Diaconu CAROL DAVILA UNIVERSITY OF MEDICINE AND PHARMACY, CLINICAL EMERGENCY HOSPITAL OF BUCHAREST, INTERNAL MEDICINE CLINIC, BUCHAREST, ROMANIA Constantin Stefani CAROL DAVILA UNIVERSITY OF MEDICINE AND PHARMACY, DEPARTMENT OF FAMILY MEDICINE, BUCHAREST, ROMANIA Ana Maria Alexandra Stanescu CAROL DAVILA UNIVERSITY OF MEDICINE AND PHARMACY, DEPARTMENT OF FAMILY MEDICINE, BUCHAREST, ROMANIA Alexandra Totan FAollowCULT thisY OF and DEN additionalTAL MEDICINE, works at:DEP https:/ARTMEN/scholarT OF.v BIOCHEMISTRalpo.edu/jmmsY, BUCHAREST, ROMANIA Part of the Obstetrics and Gynecology Commons, Oncology Commons, and the Palliative Care SeeCommons next page for additional authors Recommended Citation Miricescu, Daniela; Diaconu, Camelia Cristina; Stefani, Constantin; Stanescu, Ana Maria Alexandra; Totan, Alexandra; Rusu, Ioana Ruxandra; Bratu, Ovidiu Gabriel; Spinu, Dan; and Greabu, Maria (2020) "The Serine/ Threonine Protein Kinase (Akt)/ Protein Kinase B (PkB) Signaling Pathway in Breast Cancer," Journal of Mind and Medical Sciences: Vol. 7 : Iss. 1 , Article 7. DOI: 10.22543/7674.71.P3439 Available at: https://scholar.valpo.edu/jmms/vol7/iss1/7 This Review Article is brought to you for free and open access by ValpoScholar. It has been accepted for inclusion in Journal
    [Show full text]
  • Differential Regulation of Telomeric Complex by BCR-ABL1 Kinase In
    G C A T T A C G G C A T genes Article Differential Regulation of Telomeric Complex by BCR-ABL1 Kinase in Human Cellular Models of Chronic Myeloid Leukemia—From Single Cell Analysis to Next-Generation Sequencing 1,2, 2,3, 3 2,3 Anna Deregowska y, Monika Pepek y, Katarzyna Pruszczyk , Marcin M. Machnicki , Maciej Wnuk 1,* and Tomasz Stoklosa 4,* 1 Department of Biotechnology, University of Rzeszow, Pigonia 1, 35-310 Rzeszów, Poland; [email protected] 2 Postgraduate School of Molecular Medicine, Medical University of Warsaw, Trojdena 2a, 02-091 Warsaw, Poland; [email protected] (M.P.); [email protected] (M.M.M.) 3 Department of Immunology, Medical University of Warsaw, Nielubowicza 5, 02-097 Warsaw, Poland; [email protected] 4 Department of Tumor Biology and Genetics, Medical University of Warsaw, Pawinskiego 7, 02-106 Warsaw, Poland * Correspondence: [email protected] (M.W.); [email protected] (T.S.); Tel.: +4-822-599-2199 (T.S.) These authors contributed equally to this work. y Received: 8 August 2020; Accepted: 25 September 2020; Published: 29 September 2020 Abstract: Telomeres are specialized nucleoprotein complexes, localized at the physical ends of chromosomes, that contribute to the maintenance of genome stability. One of the features of chronic myeloid leukemia (CML) cells is a reduction in telomere length which may result in increased genomic instability and progression of the disease. Aberrant telomere maintenance in CML is not fully understood and other mechanisms such as the alternative lengthening of telomeres (ALT) are involved. In this work, we employed five BCR-ABL1-positive cell lines, namely K562, KU-812, LAMA-84, MEG-A2, and MOLM-1, commonly used in the laboratories to study the link between mutation, copy number, and expression of telomere maintenance genes with the expression, copy number, and activity of BCR-ABL1.
    [Show full text]