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https://www.readbyqxmd.com/read/29343574/novel-human-polyomavirus-non-coding-control-regions-differ-in-bi-directional-gene-expression-according-to-host-cell-large-t-antigen-expression-and-clinically-occurring-rearrangements
#1
Elvis T Ajuh, Zongsong Wu, Emma Kraus, Fabian H Weissbach, Tobias Bethge, Rainer Gosert, Nicole Fischer, Hans H Hirsch
Human polyomavirus (HPyV) DNA genomes contain three regions denoted early viral gene region (EVGR) encoding the regulatory T-antigens and one microRNA, late viral gene region (LVGR) encoding the structural Vp capsid proteins, and non-coding control region (NCCR). The NCCR harbours the origin of viral genome replication and bi-directional promoter/enhancer functions governing EVGR- and LVGR-expression on opposite DNA strands. Despite principle similarities, HPyV-NCCRs differ in length, sequence, and architecture...
January 17, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29321332/erk-is-a-critical-regulator-of-jc-polyomavirus-infection
#2
Jeanne K DuShane, Michael P Wilczek, Colleen L Mayberry, Melissa S Maginnis
The human JC polyomavirus (JCPyV) infects the majority of the population worldwide and presents as an asymptomatic, persistent infection in the kidney. In individuals who are immunocompromised, JCPyV can become reactivated and cause a lytic infection in the central nervous system resulting in the fatal, demyelinating disease progressive multifocal leukoencephalopathy (PML). Infection is initiated by interactions between the capsid protein viral protein 1 (VP1) and the α2,6-linked sialic acid on LSTc, while JCPyV internalization is facilitated by 5-hydroxytryptamine 2 receptors (5-HT2Rs)...
January 10, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29321319/non-structural-protein-%C3%AF-1s-is-required-for-optimal-reovirus-protein-expression
#3
Matthew B Phillips, Johnasha D Stuart, Emily J Simon, Karl W Boehme
Reovirus non-structural protein σ1s is required for the establishment of viremia and hematogenous viral dissemination. However, the function of the σ1s protein during the reovirus replication cycle is not known. In this study, we found that σ1s was required for efficient reovirus replication in SV40-immortalized endothelial cells (SVECs), mouse embryonic fibroblasts, human umbilical vein endothelial cells (HUVECs), and T84 human colonic epithelial cells. In each of these cell lines, wild type reovirus produced substantially higher viral titers than a σ1s-deficient mutant...
January 10, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29301535/inhibition-of-histone-deacetylase-activity-rescues-inflammatory-cystic-fibrosis-lung-disease-by-modulating-innate-and-adaptive-immune-responses
#4
Manish Bodas, Steven Mazur, Taehong Min, Neeraj Vij
BACKGROUND: Chronic lung disease resulting from dysfunctional cystic fibrosis transmembrane conductance regulator (CFTR) and NFκB-mediated neutrophilic-inflammation forms the basis of CF-related mortality. Here we aimed to evaluate if HDAC inhibition controls Pseudomonas-aeruginosa-lipopolysaccharide (Pa-LPS) induced airway inflammation and CF-lung disease. METHODS: For in vitro experiments, HEK293-cells were transfected with IL-8 or NFκB-firefly luciferase, and SV40-renilla- luciferase reporter constructs or ΔF508-CFTR-pCEP, followed by treatment with suberoylanilide hydroxamic acid (SAHA), Trichostatin-A (TSA) and/or TNFα...
January 4, 2018: Respiratory Research
https://www.readbyqxmd.com/read/29284117/knockdown-of-ampk%C3%AE-decreases-atm-expression-and-increases-radiosensitivity-under-hypoxia-and-nutrient-starvation-in-an-sv40-transformed-human-fibroblast-cell-line-lm217
#5
Yasuhiko Murata, Takuma Hashimoto, Yusuke Urushihara, Soichiro Shiga, Kazuya Takeda, Keiichi Jingu, Yoshio Hosoi
BACKGROUND: Presence of unperfused regions containing cells under hypoxia and nutrient starvation contributes to radioresistance in solid human tumors. It is well known that hypoxia causes cellular radioresistance, but little is known about the effects of nutrient starvation on radiosensitivity. We have reported that nutrient starvation induced decrease of mTORC1 activity and decrease of radiosensitivity in an SV40-transformed human fibroblast cell line, LM217, and that nutrient starvation induced increase of mTORC1 activity and increase of radiosensitivity in human liver cancer cell lines, HepG2 and HuH6 (Murata et al...
December 25, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29282022/chromosomal-instability-induced-by-increased-birc5-survivin-levels-affects-tumorigenicity-of-glioma-cells
#6
Marina Conde, Susanne Michen, Ralf Wiedemuth, Barbara Klink, Evelin Schröck, Gabriele Schackert, Achim Temme
BACKGROUND: Survivin, belonging to the inhibitor of apoptosis (IAP) gene family, is abundantly expressed in tumors. It has been hypothesized that Survivin facilitates carcinogenesis by inhibition of apoptosis resulting in improved survival of tumorigenic progeny. Additionally, Survivin plays an essential role during mitosis. Together with its molecular partners Aurora B, Borealin and inner centromere protein it secures bipolar chromosome segregation. However, whether increased Survivin levels contribute to progression of tumors by inducing chromosomal instability remains unclear...
December 28, 2017: BMC Cancer
https://www.readbyqxmd.com/read/29244783/using-a-commonly-down-regulated-cytomegalovirus-cmv-promoter-for-high-level-expression-of-ectopic-gene-in-a-human-b-lymphoma-cell-line
#7
Xiaojun Yu, Wei Geng, Hongchuan Zhao, Guobin Wang, Yijun Zhao, Zhe Zhu, Xiaoping Geng
BACKGROUND Vectors are widely used to drive gene expression using a promoter. However, not all promoters are able to drive ectopic gene expression efficiently, including CMV promoter. Here, we report our data using CMV promoter for high-level gene expression in a B lymphoma cell line DG75. MATERIAL AND METHODS A plasmid (pcDNA3.1(+)) containing the CD21 gene driven under CMV promoter was constructed. The plasmid was stably transfected into a human B lymphoma cell line DG75 for cellular surface CD21 expression, and flow cytometry was used to monitor CD21 expression...
December 15, 2017: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
https://www.readbyqxmd.com/read/29196255/hypermethylation-of-pgcp-gene-is-associated-with-human-bronchial-epithelial-cells-immortalization
#8
Chen Gao, Xiumei Xing, Zhini He, Shen Chen, Shan Wang, Qingye Li, Ping Guo, Haiyan Zhang, Huiyao Li, Liping Chen, Qing Wang, Jian Zhao, Yongmei Xiao, Wen Chen, Daochuan Li
Cell immortalization is the initial step for cancer development. To identify the differentially expressed genes regulated by DNA methylation over the course of human primary bronchial epithelial cell (HPBECs) immortalization, an immortalized HBE cell line (HBETT) was generated via introduction of an SV40 LT and a catalytic subunit of human telomerase reverse transcriptase (hTERT) into the HPBECs. Microarrays of mRNA and DNA methylation were performed to compare the transcriptomes and DNA methylomes between these two types of cells...
November 28, 2017: Gene
https://www.readbyqxmd.com/read/29167345/merkel-cell-polyomavirus-infection-of-animal-dermal-fibroblasts
#9
Wei Liu, Nathan A Krump, Margo MacDonald, Jianxin You
Merkel cell polyomavirus (MCPyV) is the first polyomavirus to be associated with human cancer. Mechanistic studies attempting to fully elucidate MCPyV oncogenic mechanisms have been hampered by the lack of animal models for MCPyV infection. In this study, we examined the infectability of MCPyV-GFP pseudovirus, MCPyV recombinant virion, and several MCPyV chimeric viruses in dermal fibroblasts isolated from various model animals, including mouse (Mus musculus), rabbit (Oryctolagus cuniculus), rat (Rattus norvegicus), chimpanzee (Pan troglodytes), rhesus macaque (Macaca mulatta), patas monkey (Erythrocebus patas), common woolly monkey (Lagothrix lagotricha), red-chested mustached tamarin (Saguinus labiatus), and tree shrew (Tupaia Belangeri)...
November 22, 2017: Journal of Virology
https://www.readbyqxmd.com/read/29167311/the-human-ctf4-orthologue-and-1-interacts-with-dna-polymerase-%C3%AE-primase-via-its-unique-c-terminal-hmg-box
#10
Mairi L Kilkenny, Aline C Simon, Jack Mainwaring, David Wirthensohn, Sandro Holzer, Luca Pellegrini
A dynamic multi-protein assembly known as the replisome is responsible for DNA synthesis in eukaryotic cells. In yeast, the hub protein Ctf4 bridges DNA helicase and DNA polymerase and recruits factors with roles in metabolic processes coupled to DNA replication. An important question in DNA replication is the extent to which the molecular architecture of the replisome is conserved between yeast and higher eukaryotes. Here, we describe the biochemical basis for the interaction of the human CTF4-orthologue AND-1 with DNA polymerase α (Pol α)/primase, the replicative polymerase that initiates DNA synthesis...
November 2017: Open Biology
https://www.readbyqxmd.com/read/29155705/identification-and-characterization-of-metastatic-factors-by-gene-transfer-into-the-novel-rip-tag-rip-tva-murine-model
#11
George Zhang, Yudan Chi, Yi-Chieh Nancy Du
Metastatic cancer accounts for 90% of deaths in patients with solid tumors. There is an urgent need to better understand the drivers of cancer metastasis and to identify novel therapeutic targets. To investigate molecular events that drive the progression from primary cancer to metastasis, we have developed a bitransgenic mouse model, RIP-Tag; RIP-tva. In this mouse model, the rat insulin promoter (RIP) drives the expression of the SV40 T antigen (Tag) and the receptor for subgroup A avian leukosis virus (tva) in pancreatic β cells...
October 16, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29107597/naked-mole-rat-cells-have-a-stable-epigenome-that-resists-ipsc%C3%A2-reprogramming
#12
Li Tan, Zhonghe Ke, Gregory Tombline, Nicholas Macoretta, Kevin Hayes, Xiao Tian, Ruitu Lv, Julia Ablaeva, Michael Gilbert, Natarajan V Bhanu, Zuo-Fei Yuan, Benjamin A Garcia, Yujiang G Shi, Yang Shi, Andrei Seluanov, Vera Gorbunova
Naked mole rat (NMR) is a valuable model for aging and cancer research due to its exceptional longevity and cancer resistance. We observed that the reprogramming efficiency of NMR fibroblasts in response to OSKM was drastically lower than that of mouse fibroblasts. Expression of SV40 LargeT antigen (LT) dramatically improved reprogramming of NMR fibroblasts. Inactivation of Rb alone, but not p53, was sufficient to improve reprogramming efficiency, suggesting that NMR chromatin may be refractory to reprogramming...
November 14, 2017: Stem Cell Reports
https://www.readbyqxmd.com/read/29070569/dexamethasone-enhances-vasopressin-induced-aquaporin-2-gene-expression-in-the-mpkccd-cells
#13
Kuang-Ting Kuo, Chan-Wei Yang, Ming-Jiun Yu
The cortical collecting duct cell (mpkCCD) has been an instrumental cell model for studying vasopressin-mediated aquaporin-2 regulation. This cell line was first developed by the Vandewalle's group from a transgenic mouse carrying the transforming SV40 antigens driven by the pyruvate kinase promoter. To immortalize the cells, four hormone supplements (dexamethasone, epidermal growth factor, insulin and triiodothyronine) were used to enhance SV40 antigen expression; however, these hormones appear to have various effects on aquaporin-2 gene expression in the cells...
October 25, 2017: American Journal of Physiology. Renal Physiology
https://www.readbyqxmd.com/read/29017784/arginine-rich-cross-linking-peptides-with-different-sv40-nuclear-localization-signal-content-as-vectors-for-intranuclear-dna-delivery
#14
Mariia Bogacheva, Anna Egorova, Anna Slita, Marianna Maretina, Vladislav Baranov, Anton Kiselev
The major barriers for intracellular DNA transportation by cationic polymers are their toxicity, poor endosomal escape and inefficient nuclear uptake. Therefore, we designed novel modular peptide-based carriers modified with SV40 nuclear localization signal (NLS). Core peptide consists of arginine, histidine and cysteine residues for DNA condensation, endosomal escape promotion and interpeptide cross-linking, respectively. We investigated three polyplexes with different NLS content (10 mol%, 50 mol% and 90 mol% of SV40 NLS) as vectors for intranuclear DNA delivery...
October 3, 2017: Bioorganic & Medicinal Chemistry Letters
https://www.readbyqxmd.com/read/28974621/topoisomerase-1-inhibition-promotes-cyclic-gmp-amp-synthase-dependent-antiviral-responses
#15
Geneviève Pépin, Charlotte Nejad, Jonathan Ferrand, Belinda J Thomas, H James Stunden, Elaine Sanij, Chwan-Hong Foo, Cameron R Stewart, Jason E Cain, Philip G Bardin, Bryan R G Williams, Michael P Gantier
Inflammatory responses, while essential for pathogen clearance, can also be deleterious to the host. Chemical inhibition of topoisomerase 1 (Top1) by low-dose camptothecin (CPT) can suppress transcriptional induction of antiviral and inflammatory genes and protect animals from excessive and damaging inflammatory responses. We describe the unexpected finding that minor DNA damage from topoisomerase 1 inhibition with low-dose CPT can trigger a strong antiviral immune response through cyclic GMP-AMP synthase (cGAS) detection of cytoplasmic DNA...
October 3, 2017: MBio
https://www.readbyqxmd.com/read/28883819/immunogenicity-of-dna-vaccine-against-h5n1-containing-extended-kappa-b-site-in-vivo-study-in-mice-and-chickens
#16
Patrycja Redkiewicz, Anna Stachyra, Róz A Sawicka, Katarzyna Bocian, Anna Góra-Sochacka, Piotr Kosson, Agnieszka Sirko
Influenza is one of the most important illnesses in the modern world, causing great public health losses each year due to the lack of medication and broadly protective, long-lasting vaccines. The development of highly immunogenic and safe vaccines is currently one of the major problems encountered in efficient influenza prevention. DNA vaccines represent a novel and powerful alternative to the conventional vaccine approaches. To improve the efficacy of the DNA vaccine against influenza H5N1, we inserted three repeated kappa B (κB) motifs, separated by a 5-bp nucleotide spacer, upstream of the cytomegalovirus promoter and downstream of the SV40 late polyadenylation signal...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28747493/role-of-viral-hemorrhagic-septicemia-virus-matrix-m-protein-in-suppressing-host-transcription
#17
Qi Ke, Wade Weaver, Adam Pore, Bartolomeo Gorgoglione, Julia Halo Wildschutte, Peng Xiao, Brian S Shepherd, Allyn Spear, Krishnamurthy Malathi, Carol A Stepien, Vikram N Vakharia, Douglas W Leaman
Viral hemorrhagic septicemia virus (VHSV) is a pathogenic fish rhabdovirus found in discrete locales throughout the Northern Hemisphere. VHSV infection of fish cells leads to upregulation of the host's virus detection response, but the virus quickly suppresses interferon (IFN) production and antiviral gene expression. By systematically screening each of the six VHSV structural and nonstructural genes, we identified matrix protein (M) as the virus' most potent antihost protein. Only M of VHSV genotype IV sublineage b (VHSV-IVb) suppressed mitochondrial antiviral signaling protein (MAVS) and type I IFN-induced gene expression in a dose-dependent manner...
October 1, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28668116/macrobac-new-technologies-for-robust-and-efficient-large-scale-production-of-recombinant-multiprotein-complexes
#18
Scott D Gradia, Justin P Ishida, Miaw-Sheue Tsai, Chris Jeans, John A Tainer, Jill O Fuss
Recombinant expression of large, multiprotein complexes is essential and often rate limiting for determining structural, biophysical, and biochemical properties of DNA repair, replication, transcription, and other key cellular processes. Baculovirus-infected insect cell expression systems are especially well suited for producing large, human proteins recombinantly, and multigene baculovirus systems have facilitated studies of multiprotein complexes. In this chapter, we describe a multigene baculovirus system called MacroBac that uses a Biobricks-type assembly method based on restriction and ligation (Series 11) or ligation-independent cloning (Series 438)...
2017: Methods in Enzymology
https://www.readbyqxmd.com/read/28664456/pancreas-specific-expression-of-oncogenes-in-a-porcine-model
#19
Martin F Berthelsen, Morten M Callesen, Tanja S Østergaard, Ying Liu, Rong Li, Henrik Callesen, Frederik Dagnæs-Hansen, Stephen Hamilton-Dutoit, Jannik E Jakobsen, Martin K Thomsen
Pancreatic cancer is the fourth leading course of cancer death and early detection of the disease is crucial for successful treatment. However, pancreatic cancer is difficult to detect in its earliest stages and once symptoms appear, the cancer has often progressed beyond possibility for curing. Research into the disease has been hampered by the lack of good models. We have generated a porcine model of pancreatic cancer with use of transgenic overexpression of an oncogene cassette containing MYC, KRAS (G12D) and SV40 LT...
October 2017: Transgenic Research
https://www.readbyqxmd.com/read/28645582/improved-pfastbac%C3%A2-donor-plasmid-vectors-for-higher-protein-production-using-the-bac-to-bac%C3%A2-baculovirus-expression-vector-system
#20
Hui Shang, Tyler A Garretson, C M Senthil Kumar, Robert F Dieter, Xiao-Wen Cheng
The Autographa californica multiple nucleopolyhedrovirus (AcMNPV)-based Bac-to-Bac(®) expression system consists of a bacmid and five pFastBac™ donor transfer vectors. It has been widely used for eukaryotic gene expression in insect cells to elucidate gene function in biotechnology laboratories. The pFastBac™ vectors contain a 50bp AcMNPV polyhedrin (polh) promoter and a 127bp SV40 polyadenylation (pA) signal for cloning a gene of interest into the bacmid, resulting in unsolved lower gene expression levels than the wild type (wt) AcMNPV in insect cells...
June 20, 2017: Journal of Biotechnology
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