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https://www.readbyqxmd.com/read/28883819/immunogenicity-of-dna-vaccine-against-h5n1-containing-extended-kappa-b-site-in-vivo-study-in-mice-and-chickens
#1
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
#2
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
#3
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
#4
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...
June 29, 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
#5
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
https://www.readbyqxmd.com/read/28636317/downsizing-proto-oncogene-cfos-to-short-helix-constrained-peptides-that-bind-jun
#6
Daniel Baxter, Samuel R Perry, Timothy A Hill, W Mei Kok, Nathan R Zaccai, R Leo Brady, David P Fairlie, Jody M Mason
The oncogenic transcription factor activator protein-1 (AP-1) is a DNA-binding protein that assembles through dimerization of Fos and Jun protein subunits, their leucine-rich helical sequences entwining into a coiled-coil structure. This study reports on downsizing the proto-oncogene cFos protein (380 residues) to shorter peptides (37-25 residues) modified with helix-inducing constraints to enhance binding to Jun. A crystal structure is reported for a 37-residue Fos-derived peptide (FosW) bound to Jun. This guided iterative downsizing of FosW to shorter peptide sequences that were constrained into stable water-soluble α-helices by connecting amino acid side chains to form cyclic pentapeptide components...
June 21, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/28614383/regulated-erlin-dependent-release-of-the-b12-transmembrane-j-protein-promotes-er-membrane-penetration-of-a-non-enveloped-virus
#7
Takamasa Inoue, Billy Tsai
The molecular mechanism by which non-enveloped viruses penetrate biological membranes remains enigmatic. The non-enveloped polyomavirus SV40 penetrates the endoplasmic reticulum (ER) membrane to reach the cytosol and cause infection. We previously demonstrated that SV40 creates its own membrane penetration structure by mobilizing select transmembrane proteins to distinct puncta in the ER membrane called foci that likely function as the cytosol entry sites. How these ER membrane proteins reorganize into the foci is unknown...
June 2017: PLoS Pathogens
https://www.readbyqxmd.com/read/28537258/exploiting-the-kinesin-1-molecular-motor-to-generate-a-virus-membrane-penetration-site
#8
Madhu Sudhan Ravindran, Martin F Engelke, Kristen J Verhey, Billy Tsai
Viruses exploit cellular machineries to penetrate a host membrane and cause infection, a process that remains enigmatic for non-enveloped viruses. Here we probe how the non-enveloped polyomavirus SV40 penetrates the endoplasmic reticulum (ER) membrane to reach the cytosol, a crucial infection step. We find that the microtubule-based motor kinesin-1 is recruited to the ER membrane by binding to the transmembrane J-protein B14. Strikingly, this motor facilitates SV40 ER-to-cytosol transport by constructing a penetration site on the ER membrane called a 'focus'...
May 24, 2017: Nature Communications
https://www.readbyqxmd.com/read/28507505/protein-protein-interaction-among-the-foxp-family-members-and-their-regulation-of-two-target-genes-vldlr-and-cntnap2-in-the-zebra-finch-song-system
#9
Ezequiel Mendoza, Constance Scharff
The Forkhead transcription factor FOXP2 is implicated in speech perception and production. The avian homolog, FoxP2 contributes to song learning and production in birds. In human cell lines, transcriptional activity of FOXP2 requires homo-dimerization or dimerization with paralogs FOXP1 or FOXP4. Whether FoxP dimerization occurs in the brain is unknown. We recently showed that FoxP1, FoxP2 and FoxP4 (FoxP1/2/4) proteins are co-expressed in neurons of Area X, a song control region in zebra finches. We now report on dimer- and oligomerization of zebra finch FoxPs and how this affects transcription...
2017: Frontiers in Molecular Neuroscience
https://www.readbyqxmd.com/read/28465536/metformin-disrupts-malignant-behavior-of-oral-squamous-cell-carcinoma-via-a-novel-signaling-involving-late-sv40-factor-aurora-a
#10
Chang-Han Chen, Hsin-Ting Tsai, Hui-Ching Chuang, Li-Yen Shiu, Li-Jen Su, Tai-Jan Chiu, Sheng-Dean Luo, Fu-Min Fang, Chao-Cheng Huang, Chih-Yen Chien
Conventional therapeutic processes in patient with OSCC are associated with several unfavorable effects leading to patients with poor survival rate. Metformin has been shown to protect against a variety of specific diseases, including cancer. However, the precise roles and mechanisms underlying the therapeutic effects of metformin on OSCC remain elusive. In the current study, in vitro and xenograft model experiments revealed that metformin inhibited growth and metastasis of oral cancer cells. Importantly, metformin-restrained tumorigenesis of oral cancer was accompanied with strong decrease of both Aurora-A and Late SV40 Factor (LSF) expressions...
May 2, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28424574/present-molecular-limitations-of-on-bipolar-cell-targeted-gene-therapy
#11
Michiel van Wyk, Elmar C Hulliger, Lara Girod, Andreas Ebneter, Sonja Kleinlogel
Recent studies have demonstrated the safety and efficacy of ocular gene therapy based on adeno-associated viral vectors (AAVs). Accordingly, a surge in promising new gene therapies is entering clinical trials, including the first optogenetic therapy for vision restoration. To date, optogenetic therapies for vision restoration target either the retinal ganglion cells (GCs) or presynaptic ON-bipolar cells (OBCs). Initiating light responses at the level of the OBCs has significant advantages over optogenetic activation of GCs...
2017: Frontiers in Neuroscience
https://www.readbyqxmd.com/read/28383556/synovial-cell-death-is-regulated-by-tnf-%C3%AE-induced-expression-of-b-cell-activating-factor-through-an-erk-dependent-increase-in-hypoxia-inducible-factor-1%C3%AE
#12
Jae-Wook Lee, Jiyoung Lee, Sung Hee Um, Eun-Yi Moon
B-cell activating factor (BAFF) has a role in the maturation and maintenance of B cells and is associated with rheumatoid arthritis (RA). Here, we investigated whether tumor necrosis factor (TNF)-α-induced BAFF expression controls the survival of fibroblast-like synoviocytes (FLS) and whether their survival can be regulated by TNF-α-mediated upregulation of hypoxia-inducible factor (HIF)-1α using MH7A synovial cells transfected with the SV40 T antigen. More TNF-α-treated cells died compared with the control...
April 6, 2017: Cell Death & Disease
https://www.readbyqxmd.com/read/28356524/sgta-dependent-regulation-of-hsc70-promotes-cytosol-entry-of-simian-virus-40-from-the-endoplasmic-reticulum
#13
Allison Dupzyk, Jeffrey M Williams, Parikshit Bagchi, Takamasa Inoue, Billy Tsai
Membrane penetration by nonenveloped viruses remains enigmatic. In the case of the nonenveloped polyomavirus simian virus 40 (SV40), the virus penetrates the endoplasmic reticulum (ER) membrane to reach the cytosol and then traffics to the nucleus to cause infection. We previously demonstrated that the cytosolic Hsc70-SGTA-Hsp105 complex is tethered to the ER membrane, where Hsp105 and SGTA facilitate the extraction of SV40 from the ER and transport of the virus into the cytosol. We now find that Hsc70 also ejects SV40 from the ER into the cytosol in a step regulated by SGTA...
June 15, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28352540/promoter-transgene-and-cell-line-effects-in-the-transfection-of-mammalian-cells-using-pdmaema-based-nano-stars
#14
Alexander Raup, Valérie Jérôme, Ruth Freitag, Christopher V Synatschke, Axel H E Müller
Non-viral transfection protocols are typically optimized using standard cells and reporter proteins, potentially underestimating cellular or transgene effects. Here such effects were studied for two human (Jurkat, HEK-293) and two rodent (CHO-K1, L929) cell lines and three fluorescent reporter proteins. Expression of the enhanced green fluorescent protein (EGFP) was studied under the control of the human elongation factor 1 alpha promoter and three viral promoters (SV40, SV40/enhancer, CMV), that of ZsYellow1 (yellow fluorescence) and mCherry (red fluorescence) for the CMV promoter...
September 2016: Biotechnology Reports
https://www.readbyqxmd.com/read/28216629/matrix-attachment-region-combinations-increase-transgene-expression-in-transfected-chinese-hamster-ovary-cells
#15
Chun-Peng Zhao, Xiao Guo, Si-Jia Chen, Chang-Zheng Li, Yun Yang, Jun-He Zhang, Shao-Nan Chen, Yan-Long Jia, Tian-Yun Wang
Matrix attachment regions (MARs) are cis-acting DNA elements that can increase transgene expression levels in a CHO cell expression system. To investigate the effects of MAR combinations on transgene expression and the underlying regulatory mechanisms, we generated constructs in which the enhanced green fluorescent protein (eGFP) gene flanked by different combinations of human β-interferon and β-globin MAR (iMAR and gMAR, respectively), which was driven by the cytomegalovirus (CMV) or simian virus (SV) 40 promoter...
February 20, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28210900/false-responses-of-renilla-luciferase-reporter-control-to-nuclear-receptor-tr4
#16
Dongyun Zhang, Sam S Atlasi, Krishna K Patel, Zihao Zhuang, Anthony P Heaney
Renilla luciferase reporter is a widely used internal control in dual luciferase reporter assay system, where its transcription is driven by a constitutively active promoter. However, the authenticity of the Renilla luciferase response in some experimental settings has recently been questioned. Testicular receptor 4 (TR4, also known as NR2C2) belongs to the subfamily 2 of nuclear receptors. TR4 binds to a direct repeat regulatory element in the promoter of a variety of target genes and plays a key role in tumorigenesis, lipoprotein regulation, and central nervous system development...
February 16, 2017: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/28206953/zinc-finger-protein-zfp36l1-promotes-osteoblastic-differentiation-but-represses-adipogenic-differentiation-of-mouse-multipotent-cells
#17
Kuo-Yun Tseng, Yi-Hsuan Chen, Shankung Lin
Zinc finger protein 36, C3H type-like 1 (ZFP36L1) is a member of the tristetraprolin (TTP) family and its role in the aging-related bone loss is currently unknown. We present evidence that ZFP36L1 expression in rat femurs and bone marrow mesenchymal stem cells (bmMSCs) was down-regulated with aging. ZFP36L1 knockdown decreased osteoblastic differentiation of MC3T3-E1 and C3H10T1/2 cells, and increased adipogenic differentiation of 3T3-L1 and C3H10T1/2 cells, whereas ZFP36L1 overexpression did the opposite. The finding that ZFP36L1 overexpression enhanced osteoblastic and repressed adipogenic differentiation was also corroborated by ex vivo experiments...
March 28, 2017: Oncotarget
https://www.readbyqxmd.com/read/28099942/bigh3-silencing-increases-retinoblastoma-tumor-growth-in-the-murine-sv40-tag-rb-model
#18
Nathalie Allaman-Pillet, Anne Oberson, Daniel F Schorderet
BIGH3, a secreted protein of the extracellular matrix interacts with collagen and integrins on the cell surface. BIGH3 can have opposing functions in cancer, acting either as tumor suppressor or promoter by enhancing tumor progression and angiogenesis. In the eye, BIGH3 is expressed in the cornea and the retinal pigment epithelium and could impact on the development of retinoblastoma, the most common paediatric intraocular neoplasm. Retinoblastoma initiation requires the inactivation of both alleles of the RB1 tumor suppressor gene in the developing retina and tumor progression involves additional genomic changes...
February 28, 2017: Oncotarget
https://www.readbyqxmd.com/read/28052271/establishment-and-characterization-of-an-sv40-large-t-antigen-transduced-porcine-colonic-epithelial-cell-line
#19
Bastian Kaiser, Martina Böttner, Thilo Wedel, Ronald M Brunner, Tom Goldammer, Szilvia Lesko, Gotthold Gäbel, Alexander Gleich, Helga Pfannkuche
Continuous cell lines have become indispensable tools that have enabled investigations into cellular mechanisms by increasing experimental reproducibility and sample availability, and decreasing the use of experimental animals. To facilitate studies of epithelial barrier function of the porcine colon, we aimed to establish an epithelial cell line with an extended replicative capacity. Cells were isolated from the proximal colon of a 3-week-old piglet and transduced using a recombinant retroviral vector construct containing the simian virus 40 large T antigen (SV40 TAg)...
2017: Cells, Tissues, Organs
https://www.readbyqxmd.com/read/28012275/emc1-dependent-stabilization-drives-membrane-penetration-of-a-partially-destabilized-non-enveloped-virus
#20
Parikshit Bagchi, Takamasa Inoue, Billy Tsai
Destabilization of a non-enveloped virus generates a membrane transport-competent viral particle. Here we probe polyomavirus SV40 endoplasmic reticulum (ER)-to-cytosol membrane transport, a decisive infection step where destabilization initiates this non-enveloped virus for membrane penetration. We find that a member of the ER membrane protein complex (EMC) called EMC1 promotes SV40 ER membrane transport and infection. Surprisingly, EMC1 does so by using its predicted transmembrane residue D961 to bind to and stabilize the membrane-embedded partially destabilized SV40, thereby preventing premature viral disassembly...
December 24, 2016: ELife
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