scholarly journals Inhibition of Thymidylate Synthetase Activity Induced in Varicella-Zoster Virus Infected Cells by (E)-5-(2-bromovinyl)-2′-deoxyuridine

1994 ◽  
Vol 5 (3) ◽  
pp. 191-194 ◽  
Author(s):  
T. Yokota ◽  
K. Konno ◽  
S. Shigeta
Keyword(s):  
Mammalian Cells ◽  
De Novo ◽  
Specific Interaction ◽  
Thymidylate Synthetase ◽  
Synthetase Activity ◽  
Focus Formation ◽  
Varicella Zoster ◽  
Key Enzyme ◽  
Infected Cells ◽  
Viral Form

The authors investigated the effect' of thymidylate synthetase (TS) on the antivariclllazoster virus (VZV) activity of ( E)5-(2-bromovinyl)-2′-deoxyuridine (BVDU). TS catalyses the conversion of deoxyuridylate (dUMP) to thymidylate (dTMP) and is a key enzyme in pyrimidine biosynthesis, providing the only source of dTMP synthesized de novo in mammalian cells. VZV encodes a specialized viral form of TS. TS activity in cells infected with VZV (TIO-VZV and TK−-VZV) increased proportionally with focus formation. From kinetic analysis using the Michaelis-Menten equation, the authors determined a Km value of 6.6μm for dUMP of TS induced in VZV-infected cells and a corresponding value from mock-infected cells of only 2.8μm. BVDU inhibited the induction of TS activity in TK+-VZV-infected cells at concentrations under ×10−3μm, but did not inhibit TS activity of TK−-VZV- or mock-infected cells at concentrations as high as 10μm. Inhibitory activity of BVDU against TS induced in TK+-VZV-infected cells appears to occur when BVDU is phosphorylated to BVDU monophosphate by viral pyrimidine kinase. These results suggest that the selective inhibitory action of BVDU on VZV replication depends on a specific interaction with both viral TK and TS. 5-Bromodeoxyuridine and 5-iododeoxyuridine inhibited TS activities induced in both VZV (TK+, TK−) and mock-infected cells. Other antiherpes compounds [i.e. 1-β-D-arabinofuranosyl- E-5-(2-bromovinyl)uracil, 9- 2-hydroxyethoxymethyl)-guanine, arabinosyladenine, and others] did not inhibit TS activity in VZV-infected cells at concentrations Of 10μm.

scholarly journals Regulation of Thymidylate Synthetase Activity in Cultured Mammalian Cells

1972 ◽  
Vol 10 (2) ◽  
pp. 471-486 ◽  
Author(s):  
ABIGAIL H. CONRAD ◽  
F. H. RUDDLE
Keyword(s):  
Enzyme Activity ◽  
Cell Division ◽  
Mammalian Cells ◽  
Actinomycin D ◽  
Specific Activity ◽  
Thymidylate Synthetase ◽  
Lag Phase ◽  
Synthetase Activity ◽  
Pronounced Increase ◽  
Culture Cycle

Changes in thymidylate synthetase specific activity in Don Chinese hamster cells grown in vitro have been examined during the culture cycle and after exposure of lag- and log-phase cultures to drugs which inhibit DNA, RNA, and protein synthesis. During the culture cycle enzyme activity was low during lag phase, rose 6- to 8-fold before log phase, fluctuated between 5.5 and 9 nmol dTMP/h/107 cells during log phase, and declined to base level during stationary phase. Puromycin prevented all increases in enzyme specific activity and caused a decrease in enzyme activity when applied to log-phase cultures. Actinomycin D prevented the initial rise in enzyme activity if applied during early lag phase but caused a pronounced increase in enzyme activity above control levels when applied during log phase. High thymidine concentration (1 mM) stopped cell division in log-phase cultures but did not alter the log-phase plateau level of thymidylate synthetase activity. Fluorodeoxyuridine stopped cell division and depressed enzyme activity to varying degrees depending upon its concentration, but at concentrations less than 10-6M enzyme activity eventually returned to normal log-phase levels and cell division resumed if puromycin was not present. Methotrexate stopped cell division and caused a 3- to 4-fold increase in enzyme activity above control levels if puromycin was not present. This increase occurred in the presence of actinomycin D but was retarded by addition of thymidine when actinomycin D was not present. These experiments suggest that the regulation of thymidylate synthetase activity in log-phase cells is complex and may involve thymidine triphosphate.

scholarly journals Viral serine palmitoyltransferase induces metabolic switch in sphingolipid biosynthesis and is required for infection of a marine alga

2016 ◽  
Vol 113 (13) ◽  
pp. E1907-E1916 ◽  
Author(s):  
Carmit Ziv ◽  
Sergey Malitsky ◽  
Alaa Othman ◽  
Shifra Ben-Dor ◽  
Yu Wei ◽  
...  
Keyword(s):  
De Novo ◽  
Serine Palmitoyltransferase ◽  
Metabolic Switch ◽  
Marine Viruses ◽  
Key Enzyme ◽  
Infected Cells ◽  
Biochemical Analyses ◽  
Biological Entities ◽  
Sphingolipid Biosynthesis

Marine viruses are the most abundant biological entities in the oceans shaping community structure and nutrient cycling. The interaction between the bloom-forming algaEmiliania huxleyiand its specific large dsDNA virus (EhV) is a major factor determining the fate of carbon in the ocean, thus serving as a key host-pathogen model system. The EhV genome encodes for a set of genes involved in the de novo sphingolipid biosynthesis, not reported in any viral genome to date. We combined detailed lipidomic and biochemical analyses to characterize the functional role of this virus-encoded pathway during lytic viral infection. We identified a major metabolic shift, mediated by differential substrate specificity of virus-encoded serine palmitoyltransferase, a key enzyme of sphingolipid biosynthesis. Consequently, unique viral glycosphingolipids, composed of unusual hydroxylated C17 sphingoid bases (t17:0) were highly enriched in the infected cells, and their synthesis was found to be essential for viral assembly. These findings uncover the biochemical bases of the virus-induced metabolic rewiring of the host sphingolipid biosynthesis during the chemical “arms race” in the ocean.

scholarly journals Dicer is involved in protection against influenza A virus infection

2007 ◽  
Vol 88 (10) ◽  
pp. 2627-2635 ◽  
Author(s):  
Alexey A. Matskevich ◽  
Karin Moelling
Keyword(s):  
Virus Infection ◽  
Influenza A Virus ◽  
Mammalian Cells ◽  
Influenza A ◽  
Virus Production ◽  
Vero Cells ◽  
Antiviral Response ◽  
Protein Levels ◽  
Infected Cells ◽  
Influenza A Virus Infection

In mammals the interferon (IFN) system is a central innate antiviral defence mechanism, while the involvement of RNA interference (RNAi) in antiviral response against RNA viruses is uncertain. Here, we tested whether RNAi is involved in the antiviral response in mammalian cells. To investigate the role of RNAi in influenza A virus-infected cells in the absence of IFN, we used Vero cells that lack IFN-α and IFN-β genes. Our results demonstrate that knockdown of a key RNAi component, Dicer, led to a modest increase of virus production and accelerated apoptosis of influenza A virus-infected cells. These effects were much weaker in the presence of IFN. The results also show that in both Vero cells and the IFN-producing alveolar epithelial A549 cell line influenza A virus targets Dicer at mRNA and protein levels. Thus, RNAi is involved in antiviral response, and Dicer is important for protection against influenza A virus infection.

scholarly journals Immune Restoration Diseases Reflect Diverse Immunopathological Mechanisms

2009 ◽  
Vol 22 (4) ◽  
pp. 651-663 ◽  
Author(s):  
Patricia Price ◽  
David M. Murdoch ◽  
Upasna Agarwal ◽  
Sharon R. Lewin ◽  
Julian H. Elliott ◽  
...  
Keyword(s):  
T Cell ◽  
Immune Responses ◽  
T Cell Activation ◽  
Cell Activation ◽  
De Novo ◽  
Granulomatous Inflammation ◽  
T Cell Response ◽  
Immune Restoration ◽  
Immunological Parameters ◽  
Varicella Zoster

SUMMARY Up to one in four patients infected with human immunodeficiency virus type 1 and given antiretroviral therapy (ART) experiences inflammatory or cellular proliferative disease associated with a preexisting opportunistic infection, which may be subclinical. These immune restoration diseases (IRD) appear to result from the restoration of immunocompetence. IRD associated with intracellular pathogens are characterized by cellular immune responses and/or granulomatous inflammation. Mycobacterial and cryptococcal IRD are attributed to a pathological overproduction of Th1 cytokines. Clinicopathological characteristics of IRD associated with viral infections suggest different pathogenic mechanisms. For example, IRD associated with varicella-zoster virus or JC polyomavirus infection correlate with a CD8 T-cell response in the central nervous system. Exacerbations or de novo presentations of hepatitis associated with hepatitis C virus (HCV) infection following ART may also reflect restoration of pathogen-specific immune responses as titers of HCV-reactive antibodies rise in parallel with liver enzymes and plasma markers of T-cell activation. Correlations between immunological parameters assessed in longitudinal sample sets and clinical presentations are required to illuminate the diverse immunological scenarios described collectively as IRD. Here we present salient clinical features and review progress toward understanding their pathogeneses.

scholarly journals CALINCA—A Novel Pipeline for the Identification of lncRNAs in Podocyte Disease

Cells ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 692
Author(s):  
Sweta Talyan ◽  
Samantha Filipów ◽  
Michael Ignarski ◽  
Magdalena Smieszek ◽  
He Chen ◽  
...  
Keyword(s):  
Cell Biology ◽  
Mammalian Cells ◽  
De Novo ◽  
Depth Information ◽  
Gene Products ◽  
Classical Analysis ◽  
Protein Coding ◽  
Bioinformatic Pipeline ◽  
Non Coding Rnas ◽  
Filtration Unit

Diseases of the renal filtration unit—the glomerulus—are the most common cause of chronic kidney disease. Podocytes are the pivotal cell type for the function of this filter and focal-segmental glomerulosclerosis (FSGS) is a classic example of a podocytopathy leading to proteinuria and glomerular scarring. Currently, no targeted treatment of FSGS is available. This lack of therapeutic strategies is explained by a limited understanding of the defects in podocyte cell biology leading to FSGS. To date, most studies in the field have focused on protein-coding genes and their gene products. However, more than 80% of all transcripts produced by mammalian cells are actually non-coding. Here, long non-coding RNAs (lncRNAs) are a relatively novel class of transcripts and have not been systematically studied in FSGS to date. The appropriate tools to facilitate lncRNA research for the renal scientific community are urgently required due to a row of challenges compared to classical analysis pipelines optimized for coding RNA expression analysis. Here, we present the bioinformatic pipeline CALINCA as a solution for this problem. CALINCA automatically analyzes datasets from murine FSGS models and quantifies both annotated and de novo assembled lncRNAs. In addition, the tool provides in-depth information on podocyte specificity of these lncRNAs, as well as evolutionary conservation and expression in human datasets making this pipeline a crucial basis to lncRNA studies in FSGS.

scholarly journals A Narrative Review on the Role of AMPK on De Novo Lipogenesis in Non-Alcoholic Fatty Liver Disease: Evidence from Human Studies

Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1822
Author(s):  
Christian von Loeffelholz ◽  
Sina M. Coldewey ◽  
Andreas L. Birkenfeld
Keyword(s):  
Liver Disease ◽  
Fatty Liver ◽  
Fatty Liver Disease ◽  
Mammalian Cells ◽  
De Novo ◽  
Adenosine Monophosphate ◽  
De Novo Lipogenesis ◽  
Alcoholic Fatty Liver ◽  
Alcoholic Fatty Liver Disease

5′AMP-activated protein kinase (AMPK) is known as metabolic sensor in mammalian cells that becomes activated by an increasing adenosine monophosphate (AMP)/adenosine triphosphate (ATP) ratio. The heterotrimeric AMPK protein comprises three subunits, each of which has multiple phosphorylation sites, playing an important role in the regulation of essential molecular pathways. By phosphorylation of downstream proteins and modulation of gene transcription AMPK functions as a master switch of energy homeostasis in tissues with high metabolic turnover, such as the liver, skeletal muscle, and adipose tissue. Regulation of AMPK under conditions of chronic caloric oversupply emerged as substantial research target to get deeper insight into the pathogenesis of non-alcoholic fatty liver disease (NAFLD). Evidence supporting the role of AMPK in NAFLD is mainly derived from preclinical cell culture and animal studies. Dysbalanced de novo lipogenesis has been identified as one of the key processes in NAFLD pathogenesis. Thus, the scope of this review is to provide an integrative overview of evidence, in particular from clinical studies and human samples, on the role of AMPK in the regulation of primarily de novo lipogenesis in human NAFLD.

Sign in / Sign up

Export Citation Format

Share Document