DOCSLIB.ORG

An Investigation of Roles for SIRT1 and Dietary Polyphenols in Modulating the Ageing Process Through DNA Methylation

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
An Investigation of Roles for SIRT1 and Dietary Polyphenols in Modulating the Ageing Process Through DNA Methylation An investigation of roles for SIRT1 and dietary polyphenols in modulating the ageing process through DNA methylation Laura Jane Ions Thesis submitted for the degree of Doctor of Philosophy Institute for Cell and Molecular Biosciences, Newcastle University, UK September 2011 Declaration I certify that this thesis is my own work, except where stated, and has not been previously submitted for a degree or any other qualification at this or any other university. Laura J Ions September 2011 Thank you to Professor Dianne Ford and Dr Luisa Wakeling whose support, guidance and enthusiasm have been invaluable over the last four years. Abstract Dietary restriction (DR) can increase lifespan across evolutionarily distinct species, from yeast to rodents. The NAD+-dependent (class III) histone deacetylase SIRT1 in mammals, and its ortholog in other species, may play a major role in this response, but may affect ‘healthspan’ (number of years of good health), rather than lifespan per se. Ageing is accompanied by changes in genome methylation, which may be causal in the ageing process. Since histones are one of the many substrates that are deacetylated by SIRT1, we hypothesised that epigenetic effects of SIRT1 activity – and specifically effects on DNA methylation, which is associated closely with histone acetylation – mediate some of the beneficial effects of DR that contribute to increased healthspan. We also propose that dietary polyphenols may act at the cellular level in a similar way. To test this hypothesis, we first investigated effects of altering SIRT1 expression, by overexpression of a transgene or siRNA-mediated knockdown, on global DNA methylation (methylation of the LINE-1 element) in the human intestinal cell line, Caco-2. We also measured effects on global DNA methylation of dietary isoflavones and resveratrol, under control conditions as well as conditions of SIRT1 overexpression. Measured effects on global DNA methylation revealed possible complex interactions between SIRT1 and these dietary polyphenols but were dependent on the assay used to measure methylation at the LINE-1 element (COBRA or pyrosequencing), so were not considered to be robust observations. We investigated factors that may affect SIRT1 expression – specifically we examined SIRT1 promoter activity in response to polyphenols, effects of promoter methylation and effects of age. Treatment of Caco-2 cells with dietary polyphenols had no effect on the SIRT1 promoter in a promoter-reporter construct. In contrast, methylation of the SIRT1 promoter reduced reporter gene expression in this model. Age did not appear to change the levels of SIRT1 protein expressed in mouse intestinal tissue when comparing young and older mice. An in silico analysis was carried out to investigate if overlaps between groups of genes compiled from published and publically-available data found to i) associate with SIRT1, ii) show altered expression in response to DR, and iii) show altered methylation with ageing were greater than expected by chance, which would support the hypothesis, and provided targets to investigate possible site-specific effects of SIRT1 on DNA methylation. Ten genes were found to fit into the ‘three way’ overlap, which was statistically greater than expected by chance. Pyrosequencing assays could be optimised for only 8 of these 10 genes, so we focused further investigations on this sub-set. Significant effects of SIRT1 overexpression and/or knockdown on methylation were observed on at least one CpG site in the I promoters of six of these genes (CDC7, EIF5, IRX3, KLF3, PTPRG, TBX3) and expression at the mRNA level of all of the eight genes (also PCYT1A and SLC39A4) was affected significantly. To gain a more comprehensive view of the extent to which DNA methylation at specific loci may be affected by SIRT1 expression levels microarray-based analysis of the methylation pattern across the genome in Caco-2 cells was carried out under conditions where SIRT1 was overexpressed or where expression was reduced by siRNA. In parallel, we measured the response to expressing SIRT1 at different levels at the level of the transcriptome. Overlaps that were statistically greater than expected by chance between these data and the lists of genes compiled from published and publically-available data used for the in silico analysis were found to exist between the complied list of genes reported to respond to dietary restriction and the set of genes we found to show altered expression in response to changing the level of SIRT1 expression and the set of genes we found to be differentially-methylated in response to reducing the level of SIRT1 expression. This observation is in broad support of our overarching hypothesis. The findings of this study indicate that effects of SIRT1 on methylation of specific genes may correspond with altered expression under conditions of dietary restriction. The data reveal a large number of gene targets for which causal links between these modifications roles in modulating the ageing process could be investigated. II Abbreviations A Adenine A260 Absorbance reading at 260nm Acetyl-CoA Acetyl-coenzyme A AceCS2 Acetyl-coenzyme A Synthetase 2 ADP Adenosine diphosphate AROS Active regulator of SIRT1 ASP Adenosine 5' phosphosulfate ATCC American Tissue Culture Collection ATP Adenosine triphosphate Bax Bcl2-associated bp Base pair BSA Bovine serum albumin C Carbon C Cytosine Caco-2 Colonic adenocarcinoma cAMP Cyclic adenosine monophosphate CCD Charged coupled device CDC7 Cell division cycle 7 CDK Cyclin dependent kinase cDNA copy DNA C/EBPα CCAAT/enhancer binding protein alpha ChREBP Carbohydrate response element binding protein COBRA Combined bisulfite restriction analysis CPRG Chlorophenol red-β-D-galactopyranoside CPS-1 Carbamoyl phosphate synthetase 1 CR Calorie restriction CREB cAMP response element binding protein CTα CTP:phosphocholine cytidylyltransferase alpha CtBP C-terminal binding protein CTP cytidine 5’-triphosphate D Aspartic acid DBC-1 Deleted in breast cancer 1 DMSO Dimethyl sulfoxide DNA Deoxyribonucleic acid DNMT1 DNA methyltransferase 1 III dNTP Deoxynucleotide triphosphate DR Dietary restriction E Glutamic acid EDTA Ethylenediaminetetraacetic acid EIF5 Eukaryotic translation initiation factor 5 ERC Extrachromosomal circles FOXO Forkhead box type O G Guanine GAP GTPase accelerating protein GDH Glutamate dehydrogenase GDI GDP dissociation inhibitor GDP Guanosine diphosphate GSTP-1 Glutathione S-transferase 1 GTP Guanosine triphosphate H Histone HAT Histone acetyltransferase HIC-1 Hypermethylated in cancer 1 HDAC Histone deacetyltransferase HeLa Henrietta Lacks IF Intermittent fasting IGF-1 Insulin-like growth factor 1 IgG Immunoglobulin G IIS Insulin/IGF-1 signalling INHAT Inhibitor of histone acetyltransferases InR Insulin receptor IRX3 Iroquois related homeobox 3 JNK c-Jun N-terminal kinase K Lysine kb Kilo bases KLF3 Krüppel-like factor 3 LB Luria Bertoni LDLR Low density lipoprotein receptor LINE-1 Long interspersed nucleotide element 1 LUMA Lumometric methylation assay MCK Muscle creatine kinase MCM2-7 Minichromosome maintenance 2-7 MeCP2 Methyl CpG binding protein 2 IV MeDIP Methylated DNA immunoprecipitation MGMT O6-methylguanine DNA methyltransferase miR Micro RNAs MMLV RT Moloney murine leukaemia virus reverse transcriptase mRNA Messenger ribonucleic acid MYH10 Myosin, heavy chain 10 NaDC3 Na+-dependent dicarboxylate cotransporter NAD(H) Nicotinamide adenine dinucleotide (hydrogen) NM II Non muscle myosin II NMHC II NM II heavy chain isoform NMNAT-1 Nicotinamide mononucleotide adenylyltransferase 1 p Phosphate p Probability PBS Phosphate buffered saline PCR Polymerase chain reaction PCYT1A Phosphate cytidylyltransferase 1 choline alpha isoform PGC-1α PPAR-gamma coactivator 1 alpha PI3K Phosphoinositide 3 kinase PIC Pre-initiation complex PPAR γ Peroxisome proliferator-activated receptor gamma PPi Pyrophosphate PTPRG Protein tyrosine phosphate receptor type G PVDF Polyvinylidene difluride RARβ Retinoic acid receptor beta r.f. Representation factor RISC RNA-induced silencing complex RNA Ribonucleic acid RNAi Ribonucleic acid interference ROS Reactive oxygen species RPTC Renal proximal tubular cells RT-PCR Reverse transcriptase-polymerase chain reaction RT-qPCR Reverse transcriptase-quantitative polymerase chain reaction S Serine SDS-PAGE Sodium dodecyl sulphate-polyacrylamide gel electrophoresis SEM Standard error of the mean SIR Silent information regulator siRNA Short interfering ribonucleic acid V SLC30A5 Solute carrier 30, member 5 SLC39A4 Solute carrier 39, member 4 SMRT Silencing mediator of retinoid and thyroid hormone receptor STAC Sirtuin activating compound T Thymine TBX3 T-box3 transcription factor TDR Transcriptional repressor domain TEMED N,N,N’,N’-Tetramethylethylenediamine tk Thymidine kinase TOR Target of rapamycin TSA Trichostatin A UCP2 Uncoupling protein 2 UV Ultra violet WAT White adipose tissue VI Contents Abstract .................................................................................................................................................... I Abbreviations ........................................................................................................................................ III Contents................................................................................................................................................ VII List of Figures ........................................................................................................................................
Load more

Recommended publications
  • The VLDL Receptor Regulates Membrane Progesterone Receptor
    © 2018. Published by The Company of Biologists Ltd | Journal of Cell Science (2018) 131, jcs212522. doi:10.1242/jcs.212522 RESEARCH ARTICLE The VLDL receptor regulates membrane progesterone receptor trafficking and non-genomic signaling Nancy Nader, Maya Dib, Raphael Courjaret, Rawad Hodeify, Raya Machaca, Johannes Graumann and Khaled Machaca* ABSTRACT the plasma membrane and interact with the classical P4 receptor, is Progesterone mediates its physiological functions through activation of nonetheless effective at mediating non-genomic P4 signaling both transcription-coupled nuclear receptors and seven-pass- (Bandyopadhyay et al., 1998; Dressing et al., 2011; Peluso et al., transmembrane progesterone receptors (mPRs), which transduce 2002). These results argued for the presence of membrane P4 the rapid non-genomic actions of progesterone by coupling to various receptors that are distinct from the nuclear P4 receptors. In 2003, the signaling modules. However, the immediate mechanisms of action Thomas laboratory identified a family of membrane progesterone downstream of mPRs remain in question. Herein, we use an untargeted receptors (mPRs) from fish ovaries (Zhu et al., 2003a,b) that belong quantitative proteomics approach to identify mPR interactors to better to the progestin and adiponectin (AdipoQ) receptor family (also define progesterone non-genomic signaling. Surprisingly, we identify named PAQ receptors). However, the signal transduction cascade the very-low-density lipoprotein receptor (VLDLR) as an mPRβ downstream of mPRs that mediates the non-genomic actions of P4 (PAQR8) partner that is required for mPRβ plasma membrane remains unclear. localization. Knocking down VLDLR abolishes non-genomic The non-genomic action of mPR and the ensuing signaling progesterone signaling, which is rescued by overexpressing VLDLR.
    [Show full text]
  • Progesterone Receptor Membrane Component 1 Promotes Survival of Human Breast Cancer Cells and the Growth of Xenograft Tumors
    Cancer Biology & Therapy ISSN: 1538-4047 (Print) 1555-8576 (Online) Journal homepage: http://www.tandfonline.com/loi/kcbt20 Progesterone receptor membrane component 1 promotes survival of human breast cancer cells and the growth of xenograft tumors Nicole C. Clark, Anne M. Friel, Cindy A. Pru, Ling Zhang, Toshi Shioda, Bo R. Rueda, John J. Peluso & James K. Pru To cite this article: Nicole C. Clark, Anne M. Friel, Cindy A. Pru, Ling Zhang, Toshi Shioda, Bo R. Rueda, John J. Peluso & James K. Pru (2016) Progesterone receptor membrane component 1 promotes survival of human breast cancer cells and the growth of xenograft tumors, Cancer Biology & Therapy, 17:3, 262-271, DOI: 10.1080/15384047.2016.1139240 To link to this article: http://dx.doi.org/10.1080/15384047.2016.1139240 Accepted author version posted online: 19 Jan 2016. Published online: 19 Jan 2016. Submit your article to this journal Article views: 49 View related articles View Crossmark data Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=kcbt20 Download by: [University of Connecticut] Date: 26 May 2016, At: 11:28 CANCER BIOLOGY & THERAPY 2016, VOL. 17, NO. 3, 262–271 http://dx.doi.org/10.1080/15384047.2016.1139240 RESEARCH PAPER Progesterone receptor membrane component 1 promotes survival of human breast cancer cells and the growth of xenograft tumors Nicole C. Clarka,*, Anne M. Frielb,*, Cindy A. Prua, Ling Zhangb, Toshi Shiodac, Bo R. Ruedab, John J. Pelusod, and James K. Prua aDepartment of Animal Sciences,
    [Show full text]
  • The Roles of Histone Deacetylase 5 and the Histone Methyltransferase Adaptor WDR5 in Myc Oncogenesis
    The Roles of Histone Deacetylase 5 and the Histone Methyltransferase Adaptor WDR5 in Myc oncogenesis By Yuting Sun This thesis is submitted in fulfilment of the requirements for the degree of Doctor of Philosophy at the University of New South Wales Children’s Cancer Institute Australia for Medical Research School of Women’s and Children’s Health, Faculty of Medicine University of New South Wales Australia August 2014 PLEASE TYPE THE UNIVERSITY OF NEW SOUTH WALES Thesis/Dissertation Sheet Surname or Family name: Sun First name: Yuting Other name/s: Abbreviation for degree as given in the University calendar: PhD School : School of·Women's and Children's Health Faculty: Faculty of Medicine Title: The Roles of Histone Deacetylase 5 and the Histone Methyltransferase Adaptor WDR5 in Myc oncogenesis. Abstract 350 words maximum: (PLEASE TYPE) N-Myc Induces neuroblastoma by regulating the expression of target genes and proteins, and N-Myc protein is degraded by Fbxw7 and NEDD4 and stabilized by Aurora A. The class lla histone deacetylase HDAC5 suppresses gene transcription, and blocks myoblast and leukaemia cell differentiation. While histone H3 lysine 4 (H3K4) trimethylation at target gene promoters is a pre-requisite for Myc· induced transcriptional activation, WDRS, as a histone H3K4 methyltransferase presenter, is required for H3K4 methylation and transcriptional activation mediated by a histone H3K4 methyltransferase complex. Here, I investigated the roles of HDAC5 and WDR5 in N-Myc overexpressing neuroblastoma. I have found that N-Myc upregulates HDAC5 protein expression, and that HDAC5 represses NEDD4 gene expression, increases Aurora A gene expression and consequently upregulates N-Myc protein expression in neuroblastoma cells.
    [Show full text]
  • Investigation of the Underlying Hub Genes and Molexular Pathogensis in Gastric Cancer by Integrated Bioinformatic Analyses
    bioRxiv preprint doi: https://doi.org/10.1101/2020.12.20.423656; this version posted December 22, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Investigation of the underlying hub genes and molexular pathogensis in gastric cancer by integrated bioinformatic analyses Basavaraj Vastrad1, Chanabasayya Vastrad*2 1. Department of Biochemistry, Basaveshwar College of Pharmacy, Gadag, Karnataka 582103, India. 2. Biostatistics and Bioinformatics, Chanabasava Nilaya, Bharthinagar, Dharwad 580001, Karanataka, India. * Chanabasayya Vastrad [email protected] Ph: +919480073398 Chanabasava Nilaya, Bharthinagar, Dharwad 580001 , Karanataka, India bioRxiv preprint doi: https://doi.org/10.1101/2020.12.20.423656; this version posted December 22, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract The high mortality rate of gastric cancer (GC) is in part due to the absence of initial disclosure of its biomarkers. The recognition of important genes associated in GC is therefore recommended to advance clinical prognosis, diagnosis and and treatment outcomes. The current investigation used the microarray dataset GSE113255 RNA seq data from the Gene Expression Omnibus database to diagnose differentially expressed genes (DEGs). Pathway and gene ontology enrichment analyses were performed, and a proteinprotein interaction network, modules, target genes - miRNA regulatory network and target genes - TF regulatory network were constructed and analyzed. Finally, validation of hub genes was performed. The 1008 DEGs identified consisted of 505 up regulated genes and 503 down regulated genes.
    [Show full text]
  • DYSREGULATION of LONG-CHAIN ACYL-Coa SYNTHETASES in CANCER and THEIR TARGETING STRATEGIES in ANTICANCER THERAPY Md Amir Hossain1, Jun Ma2, Yong Yang1 1
    Hossain et al RJLBPCS 2020 www.rjlbpcs.com Life Science Informatics Publications Original Review Article DOI: 10.26479/2020.0603.06 DYSREGULATION OF LONG-CHAIN ACYL-CoA SYNTHETASES IN CANCER AND THEIR TARGETING STRATEGIES IN ANTICANCER THERAPY Md Amir Hossain1, Jun Ma2, Yong Yang1 1. Academic Institute of Pharmaceutical Science, China Pharmaceutical University, Nanjing, China. 2. Ningxia Baoshihua Hospital, Ningxia, China. ABSTRACT: Cancer is a leading cause of death worldwide, and the number of cases globally continues to increase. Cancer is caused by certain changes in genes that are involved in controlling cell functions and cell division. Notably, metabolic dysregulation is one the hallmarks of cancer, and increases in fatty acid metabolism have been demonstrated to promote the growth and survival of a variety of cancers. In human, fatty acids either can be breakdown into acetyl-CoA through catabolic metabolism and aid in ATP generation or in the anabolic metabolism they can incorporate into triacylglycerol and phospholipid. Importantly, both of these pathways need activation of fatty acids, and the key players in this activation of fatty acids are the long-chain acyl-CoA synthetases (ACSLs) that are commonly dysregulated in cancer and associated with oncogenesis and survival. Therefore, it provides a rationale to target ACSLs in cancer. This review summarizes the current understanding of long-chain acyl-CoA synthetases in cancer and their targeting opportunities. KEYWORDS: Acyl-CoA synthetases, cancer, fatty acid, lipid metabolism. Article History: Received: April 15, 2020; Revised: May 10, 2020; Accepted: May 30, 2020. Corresponding Author: Prof. Yong Yang* Academic Institute of Pharmaceutical Science, China Pharmaceutical University, Nanjing, China.
    [Show full text]
  • Progesterone – Friend Or Foe?
    Frontiers in Neuroendocrinology 59 (2020) 100856 Contents lists available at ScienceDirect Frontiers in Neuroendocrinology journal homepage: www.elsevier.com/locate/yfrne Progesterone – Friend or foe? T ⁎ Inger Sundström-Poromaaa, , Erika Comascob, Rachael Sumnerc, Eileen Ludersd,e a Department of Women’s and Children’s Health, Uppsala University, Sweden b Department of Neuroscience, Science for Life Laboratory, Uppsala University, Uppsala, Sweden c School of Pharmacy, University of Auckland, New Zealand d School of Psychology, University of Auckland, New Zealand e Laboratory of Neuro Imaging, School of Medicine, University of Southern California, Los Angeles, USA ARTICLE INFO ABSTRACT Keywords: Estradiol is the “prototypic” sex hormone of women. Yet, women have another sex hormone, which is often Allopregnanolone disregarded: Progesterone. The goal of this article is to provide a comprehensive review on progesterone, and its Emotion metabolite allopregnanolone, emphasizing three key areas: biological properties, main functions, and effects on Hormonal contraceptives mood in women. Recent years of intensive research on progesterone and allopregnanolone have paved the way Postpartum depression for new treatment of postpartum depression. However, treatment for premenstrual syndrome and premenstrual Premenstrual dysphoric disorder dysphoric disorder as well as contraception that women can use without risking mental health problems are still Progesterone needed. As far as progesterone is concerned, we might be dealing with a two-edged sword: while its metabolite allopregnanolone has been proven useful for treatment of PPD, it may trigger negative symptoms in women with PMS and PMDD. Overall, our current knowledge on the beneficial and harmful effects of progesterone is limited and further research is imperative. Introduction 1.
    [Show full text]
  • Caracterització De L'efecte De Compostos Naturals En Models In
    Caracterització de l’efecte de compostos naturals en models in vitro i in vivo de càncer de còlon Susana Sánchez Tena ADVERTIMENT. La consulta d’aquesta tesi queda condicionada a l’acceptació de les següents condicions d'ús: La difusió d’aquesta tesi per mitjà del servei TDX (www.tdx.cat) i a través del Dipòsit Digital de la UB (diposit.ub.edu) ha estat autoritzada pels titulars dels drets de propietat intel·lectual únicament per a usos privats emmarcats en activitats d’investigació i docència. No s’autoritza la seva reproducció amb finalitats de lucre ni la seva difusió i posada a disposició des d’un lloc aliè al servei TDX ni al Dipòsit Digital de la UB. No s’autoritza la presentació del seu contingut en una finestra o marc aliè a TDX o al Dipòsit Digital de la UB (framing). Aquesta reserva de drets afecta tant al resum de presentació de la tesi com als seus continguts. En la utilització o cita de parts de la tesi és obligat indicar el nom de la persona autora. ADVERTENCIA. La consulta de esta tesis queda condicionada a la aceptación de las siguientes condiciones de uso: La difusión de esta tesis por medio del servicio TDR (www.tdx.cat) y a través del Repositorio Digital de la UB (diposit.ub.edu) ha sido autorizada por los titulares de los derechos de propiedad intelectual únicamente para usos privados enmarcados en actividades de investigación y docencia. No se autoriza su reproducción con finalidades de lucro ni su difusión y puesta a disposición desde un sitio ajeno al servicio TDR o al Repositorio Digital de la UB.
    [Show full text]
  • Flawed Phospholipid Formation Or Faulty Fatty Acid Oxidation: Determining the Cause of Mitochondrial Dysfunction in Hearts Lacking Acsl1
    FLAWED PHOSPHOLIPID FORMATION OR FAULTY FATTY ACID OXIDATION: DETERMINING THE CAUSE OF MITOCHONDRIAL DYSFUNCTION IN HEARTS LACKING ACSL1 Trisha J. Grevengoed A dissertation submitted to the faculty at the University of North Carolina at Chapel Hill in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Department of Nutrition (Biochemistry) in the School of Public Health. Chapel Hill 2015 Approved by: Rosalind A. Coleman Stephen D. Hursting Liza Makowski Leslie V. Parise Steven H. Zeisel © 2015 Trisha J. Grevengoed ALL RIGHTS RESERVED ii ABSTRACT Trisha J. Grevengoed: Fatty acid activation in cardiac mitochondria: The role of ACSL1 in phospholipid formation and remodeling, substrate switching, and autophagic flux (Under the direction of Rosalind A. Coleman) Cardiovascular disease is the number one cause of death worldwide. In the heart, mitochondria provide up to 95% of energy, with most of this energy coming from metabolism of fatty acids (FA). FA must be converted to acyl-CoAs by acyl-CoA synthetases (ACS) before entry into pathways of β- oxidation or glycerolipid synthesis. ACSL1 contributes more than 90% of total cardiac ACSL activity, and mice with an inducible knockout of ACSL1 (Acsl1T-/-) have impaired cardiac FA oxidation. The effects of loss of ACSL1 on mitochondrial respiratory function, phospholipid formation, or autophagic flux have not yet been studied. Acsl1T-/- hearts contained 3-fold more mitochondria with abnormal structure and displayed lower respiratory function. Because ACSL1 exhibited a strong substrate preference for linoleate (18:2), we investigated the composition of mitochondrial phospholipids. Acsl1T-/- hearts contained 83% less tetralinoleoyl-cardiolipin (CL), the major form present in control hearts.
    [Show full text]
  • G Protein-Coupled Receptors Function As Cell Membrane Receptors for the Steroid Hormone 20-Hydroxyecdysone Xiao-Fan Zhao
    Zhao Cell Communication and Signaling (2020) 18:146 https://doi.org/10.1186/s12964-020-00620-y REVIEW Open Access G protein-coupled receptors function as cell membrane receptors for the steroid hormone 20-hydroxyecdysone Xiao-Fan Zhao Abstract G protein-coupled receptors (GPCRs) are cell membrane receptors for various ligands. Recent studies have suggested that GPCRs transmit animal steroid hormone signals. Certain GPCRs have been shown to bind steroid hormones, for example, G protein-coupled estrogen receptor 1 (GPER1) binds estrogen in humans, and Drosophila dopamine/ecdysteroid receptor (DopEcR) binds the molting hormone 20-hydroxyecdysone (20E) in insects. This review summarizes the research progress on GPCRs as animal steroid hormone cell membrane receptors, including the nuclear and cell membrane receptors of steroid hormones in mammals and insects, the 20E signaling cascade via GPCRs, termination of 20E signaling, and the relationship between genomic action and the nongenomic action of 20E. Studies indicate that 20E induces a signal via GPCRs to regulate rapid cellular responses, including rapid Ca2+ release from the endoplasmic reticulum and influx from the extracellular medium, as well as rapid protein phosphorylation and subcellular translocation. 20E via the GPCR/Ca2+/PKC/signaling axis and the GPCR/cAMP/PKA- signaling axis regulates gene transcription by adjusting transcription complex formation and DNA binding activity. GPCRs can bind 20E in the cell membrane and after being isolated, suggesting GPCRs as cell membrane receptors of 20E. This review deepens our understanding of GPCRs as steroid hormone cell membrane receptors and the GPCR-mediated signaling pathway of 20E (20E-GPCR pathway), which will promote further study of steroid hormone signaling via GPCRs, and presents GPCRs as targets to explore new pharmaceutical materials to treat steroid hormone-related diseases or control pest insects.
    [Show full text]
  • Elucidating Biological Roles of Novel Murine Genes in Hearing Impairment in Africa
    Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 19 September 2019 doi:10.20944/preprints201909.0222.v1 Review Elucidating Biological Roles of Novel Murine Genes in Hearing Impairment in Africa Oluwafemi Gabriel Oluwole,1* Abdoulaye Yal 1,2, Edmond Wonkam1, Noluthando Manyisa1, Jack Morrice1, Gaston K. Mazanda1 and Ambroise Wonkam1* 1Division of Human Genetics, Department of Pathology, Faculty of Health Sciences, University of Cape Town, Observatory, Cape Town, South Africa. 2Department of Neurology, Point G Teaching Hospital, University of Sciences, Techniques and Technology, Bamako, Mali. *Correspondence to: [email protected]; [email protected] Abstract: The prevalence of congenital hearing impairment (HI) is highest in Africa. Estimates evaluated genetic causes to account for 31% of HI cases in Africa, but the identification of associated causative genes mutations have been challenging. In this study, we reviewed the potential roles, in humans, of 38 novel genes identified in a murine study. We gathered information from various genomic annotation databases and performed functional enrichment analysis using online resources i.e. genemania and g.proflier. Results revealed that 27/38 genes are express mostly in the brain, suggesting additional cognitive roles. Indeed, HERC1- R3250X had been associated with intellectual disability in a Moroccan family. A homozygous 216-bp deletion in KLC2 was found in two siblings of Egyptian descent with spastic paraplegia. Up to 27/38 murine genes have link to at least a disease, and the commonest mode of inheritance is autosomal recessive (n=8). Network analysis indicates that 20 other genes have intermediate and biological links to the novel genes, suggesting their possible roles in HI.
    [Show full text]
  • Open Sadiesteffens Dissertation Final.Pdf
    The Pennsylvania State University The Graduate School College of Medicine MECHANISM OF DRUG ACTION OF THE SPECIFIC CK2 INHIBITOR CX-4945 IN ACUTE MYELOID LEUKEMIA A Dissertation in Biomedical Sciences by Sadie Lynne Steffens 2015 Sadie Lynne Steffens Submitted in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy December 2015 The dissertation of Sadie Lynne Steffens was reviewed and approved* by the following: Sinisa Dovat Physician, Associate Professor of Pediatrics, Pharmacology, & Biochemistry Director, Translational Research – Four Diamonds Pediatric Cancer Research Center Dissertation Advisor Chair of Committee Barbara A. Miller Physician, Professor of Pediatrics Chief, Division of Pediatric Hematology/Oncology Sergei A. Grigoryev Professor of Biochemistry and Molecular Biology Jong K. Yun Associate Professor of Pharmacology Ralph L. Keil Associate Professor of Biochemistry and Molecular Biology Chair, Biomedical Sciences Graduate Program *Signatures are on file in the Graduate School ii ABSTRACT Acute myeloid leukemia (AML) is a malignant disease of the myeloid line of blood cells and is characterized by the rapid growth of abnormal white blood cells that accumulate in the bone marrow and interfere with the production of normal blood cells. Cytarabine and other currently available treatments for acute myeloid leukemia are highly toxic and insufficient, as more than half of all AML patients develop resistance to chemotherapeutic agents. Since AML often affects older people who are less tolerant of chemotherapy, there is need for novel, targeted, less toxic drugs in order to improve survival for this disease. Casein Kinase II (CK2) is a pro-oncogenic serine/threonine kinase that is essential for cellular proliferation.
    [Show full text]
  • Membrane Progesterone Receptor Beta (Mprβ/Paqr8) Promotes
    www.nature.com/scientificreports OPEN Membrane progesterone receptor beta (mPRβ/Paqr8) promotes progesterone-dependent neurite Received: 19 December 2016 Accepted: 30 May 2017 outgrowth in PC12 neuronal cells Published: xx xx xxxx via non-G protein-coupled receptor (GPCR) signaling Mayu Kasubuchi1, Keita Watanabe1, Kanako Hirano2, Daisuke Inoue2, Xuan Li1, Kazuya Terasawa3, Morichika Konishi4, Nobuyuki Itoh2 & Ikuo Kimura1 Recently, sex steroid membrane receptors garnered world-wide attention because they may be related to sex hormone-mediated unknown rapid non-genomic action that cannot be currently explained by their genomic action via nuclear receptors. Progesterone affects cell proliferation and survival via non- genomic effects. In this process, membrane progesterone receptors (mPRα, mPRβ, mPRγ, mPRδ, and mPRε) were identified as putative G protein-coupled receptors (GPCRs) for progesterone. However, the structure, intracellular signaling, and physiological functions of these progesterone receptors are still unclear. Here, we identify a molecular mechanism by which progesterone promotes neurite outgrowth through mPRβ (Paqr8) activation. Mouse mPRβ mRNA was specifically expressed in the central nervous system. It has an incomplete GPCR topology, presenting 6 transmembrane domains and did not exhibit typical GPCR signaling. Progesterone-dependent neurite outgrowth was exhibited by the promotion of ERK phosphorylation via mPRβ, but not via other progesterone receptors such as progesterone membrane receptor 1 (PGRMC-1) and nuclear progesterone receptor in nerve growth factor-induced neuronal PC12 cells. These findings provide new insights of regarding the non-genomic action of progesterone in the central nervous system. Steroid hormones such as corticosterone, progesterone, testosterone, and estrogen are known to exhibit their physiological effects via their specific nuclear receptors1.
    [Show full text]