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sv40 promoter

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https://www.readbyqxmd.com/read/29769335/bag2-is-a-component-of-a-cytosolic-extraction-machinery-that-promotes-membrane-penetration-of-a-nonenveloped-virus
#1
Allison Dupzyk, Billy Tsai
During entry, the nonenveloped polyomavirus (PyV) SV40 traffics from the cell surface to the endoplasmic reticulum (ER) where it penetrates the ER membrane to reach the cytosol; the virus then transports into the nucleus to cause infection. Although a coherent understanding of SV40's host entry is emerging, how the virus is ejected from the ER into the cytosol remains mysterious. Our previous analyses revealed that the cytosolic Hsc70-SGTA-Hsp105 complex binds to and extracts SV40 from the ER into the cytosol...
May 16, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29737083/-effects-of-different-promoters-and-mar-combinations-on-transgene-expression-of-recombinant-cho-cells
#2
Qin Li, Xiao-Yin Wang, Chun-Peng Zhao, Zheng-Wei Tian, Dan-Hua Xu, Tian-Yun Wang, Jun-He Zhang
OBJECTIVE: To analyze the effects of different promoters and matrix attachment region (MAR) on the expression of transgene in Chinese hamster ovary (CHO) cells. METHODS: The expression vector was constructed by the combination of beta globin MAR (gMAR) with the human cytomegalovirus immediate-early promoter (CMV-IE) and simian virus 40 (SV40) promoter. These vectors were transfected into CHO cells,after 48 h,the transient expression of enhanced green fluorescent protein (eGFP) was observed; G418 was used to screen stably transformed cell lines,and the expression level of eGFP in CHO cells was analyzed by flow cytometry...
January 2018: Sichuan da Xue Xue Bao. Yi Xue Ban, Journal of Sichuan University. Medical Science Edition
https://www.readbyqxmd.com/read/29731503/evaluation-of-teneligliptin-effects-on-transcriptional-activity-of-ppar%C3%AE-in-cell-based-assays
#3
Yasuhiro Takenaka, Ikuo Inoue, Takanari Nakano, Masaaki Ikeda, Yoshihiko Kakinuma, Yuichi Ikegami, Akira Shimada, Mitsuhiko Noda
BACKGROUND: The antidiabetic drug teneligliptin is a novel dipeptidyl peptidase-4 (DPP-4) inhibitor with a thiazolidine-specific structure. This study aimed to investigate whether teneligliptin can activate PPARγ directly and/or indirectly in cell-based assays. METHODS: Promoter assays using the reporter construct driven under the control of the SV40 promoter and the PPAR response element (PPRE) were performed. Luciferase activity was measured after a 3-day incubation of vector-transduced cells with various concentrations of teneligliptin...
2018: Journal of Nippon Medical School, Nippon Ika Daigaku Zasshi
https://www.readbyqxmd.com/read/29593037/dynein-engages-and-disassembles-cytosol-localized-sv40-to-promote-infection
#4
Madhu Sudhan Ravindran, Chelsey C Spriggs, Kristen J Verhey, Billy Tsai
During entry, polyomavirus (PyV) is endocytosed and sorts to the endoplasmic reticulum (ER) where it penetrates the ER membrane to reach the cytosol. From the cytosol, the virus moves to the nucleus to cause infection. How PyV is transported from the cytosol into the nucleus, a crucial infection step, is unclear. We found that upon reaching the cytosol, the archetype PyV SV40 recruits the cytoplasmic dynein motor, which disassembles the viral particle. This reaction enables the resulting disassembled virus to enter the nucleus to promote infection...
March 28, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29581840/interferon-beta-induces-apoptosis-in-nasopharyngeal-carcinoma-cells-via-the-trail-signaling-pathway
#5
Anna Makowska, Lora Wahab, Till Braunschweig, Nikiforos-Ioannis Kapetanakis, Christian Vokuhl, Bernd Denecke, Lian Shen, Pierre Busson, Udo Kontny
The combination of neoadjuvant chemotherapy, radiochemotherapy, and maintenance therapy with interferon beta (IFNβ) has led to superior results in the treatment of children and adolescents with nasopharyngeal carcinoma (NPC). However, nothing is known about the mechanism of the antitumor activity of IFNβ in NPC. Here, we investigate the role of IFNβ on apoptosis in NPC cells. Six NPC cell lines, one patient-derived NPC xenograft (PDX) and one SV40-transformed nasoepithelial cell line were used. Induction of apoptosis by IFNβ was measured by flow cytometric analysis of subG1-DNA-content, Hoechst 33258 staining and activation of caspase-3...
March 6, 2018: Oncotarget
https://www.readbyqxmd.com/read/29565571/t7-polymerase-expression-of-guide-rnas-in-vivo-allows-exportable-crispr-cas9-editing-in-multiple-yeast-hosts
#6
Nicholas J Morse, James M Wagner, Kevin Reed, Madan R Gopal, Lars Lauffer, Hal S Alper
Efficient guide RNA expression often limits CRISPR-Cas9 implementation in new hosts. To address this limitation in fungal systems, we demonstrate the utility of a T7 polymerase system to effectively express sgRNAs. Initially, we developed a methodology in Saccharomyces cerevisiae using a modified version of the T7 P266L mutant polymerase with an SV40 nuclear localization signal to allow guide RNA expression immediately downstream of a T7 promoter. To improve targeting efficiency, guide RNA design was found to be tolerant to three mismatches or up to three additional bases appended to the 5' end...
March 22, 2018: ACS Synthetic Biology
https://www.readbyqxmd.com/read/29560129/-hsa-mir-155-5p-drives-aneuploidy-at-early-stages-of-cellular-transformation
#7
Sara Pagotto, Angelo Veronese, Alessandra Soranno, Paola Lanuti, Mirco Di Marco, Marco Vincenzo Russo, Alice Ramassone, Marco Marchisio, Pasquale Simeone, Paolo E Guanciali Franchi, Giandomenico Palka, Renato Mariani Costantini, Carlo M Croce, Rosa Visone
Hsa-miR-155-5p (miR-155) is overexpressed in most solid and hematological malignancies. It promotes loss of genomic integrity in cancer cells by targeting genes involved in microsatellite instability and DNA repair; however, the link between miR-155 and aneuploidy has been scarcely investigated. Here we describe a novel mechanism by which miR-155 causes chromosomal instability. Using osteosarcoma cells (U2OS) and normal human dermal fibroblast (HDF), two well-established models for the study of chromosome congression, we demonstrate that miR-155 targets the spindle checkpoint proteins BUB1, CENP-F, and ZW10, thus compromising chromosome alignment at the metaphase plate...
February 27, 2018: Oncotarget
https://www.readbyqxmd.com/read/29523597/cross-talk-between-myeloid-derived-suppressor-cells-and-mast-cells-mediates-tumor-specific-immunosuppression-in-prostate-cancer
#8
Elena Jachetti, Valeria Cancila, Alice Rigoni, Lucia Bongiovanni, Barbara Cappetti, Beatrice Belmonte, Claudia Enriquez, Patrizia Casalini, Paola Ostano, Barbara Frossi, Sabina Sangaletti, Claudia Chiodoni, Giovanna Chiorino, Carlo E Pucillo, Claudio Tripodo, Mario P Colombo
Immunotherapy, including the use of checkpoint inhibitors, is a potent therapeutic approach for some cancers, but has limited success with prostate tumors, in which immune suppression is instigated by the tumor. The immunosuppressive capacity of mast cells, which promote adenocarcinoma development in the prostate, prompted our investigation on whether mast cells promote tolerance to SV40 Large-T antigen, the transforming oncogene in transgenic adenocarcinoma of the mouse prostate (TRAMP) mice. The incidence of adenocarcinoma was reduced in the offspring of a cross between TRAMP mice and mast cell-deficient KitWsh mice...
May 2018: Cancer Immunology Research
https://www.readbyqxmd.com/read/29380239/vegf-pkd-1-signaling-mediates-arteriogenic-gene-expression-and-angiogenic-responses-in-reversible-human-microvascular-endothelial-cells-with-extended-lifespan
#9
Brad Best, Patrick Moran, Bin Ren
Microvascular ECs (MVECs) are an ideal model in angiogenesis research. The aim of this study was to determine vascular endothelial growth factor (VEGF)/protein kinase D1 (PKD-1) signaling in expression of arteriogenic genes in human MVECs. To achieve this aim, we transduced specific SV40 large T antigen and telomerase into primary human dermal MVECs (HMVEC-D) to establish reversible HMVECs with extended lifespan (HMVECi-D). HMVECi-D was then exposed to VEGF/VEGF-inducer GS4012 or transduced with constitutively active protein kinase PKD-1 (PKD-CA)...
January 29, 2018: Molecular and Cellular Biochemistry
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
#10
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
#11
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 kidneys. 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 lactoseries tetrasaccharide c (LSTc), while JCPyV internalization is facilitated by 5-hydroxytryptamine 2 receptors (5-HT2 Rs)...
April 1, 2018: Journal of Virology
https://www.readbyqxmd.com/read/29321319/non-structural-protein-%C3%AF-1s-is-required-for-optimal-reovirus-protein-expression
#12
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
#13
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
#14
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...
January 22, 2018: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29282022/chromosomal-instability-induced-by-increased-birc5-survivin-levels-affects-tumorigenicity-of-glioma-cells
#15
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
#16
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
#17
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...
February 5, 2018: Gene
https://www.readbyqxmd.com/read/29167345/merkel-cell-polyomavirus-infection-of-animal-dermal-fibroblasts
#18
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's oncogenic mechanisms have been hampered by the lack of animal models for MCPyV infection. In this study, we examined the ability of MCPyV-GFP pseudovirus (containing a green fluorescent protein [GFP] reporter construct), MCPyV recombinant virions, and several MCPyV chimeric viruses to infect 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 )...
February 15, 2018: 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
#19
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
#20
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
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