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Tet proteins

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https://www.readbyqxmd.com/read/29431990/bioorthogonally-applicable-fluorogenic-cyanine-tetrazines-for-no-wash-super-resolution-imaging
#1
Gergely Knorr, Eszter Kozma, Judith Schaart, Krisztina Németh, György Török, Peter Kele
The synthesis, fluorogenic characterization and labeling application of four tetrazine-quenched cyanine probes with emission maxima in the red - far red range is reported. Fluorescence of the cyanine-cores is quenched via through-bond-energy-transfer (TBET) exerted by a bioorthogonal tetrazine unit. Upon bioorthogonal labeling reaction with cyclooctyne tagged proteins, the quenching effect ceases, thus the fluorescence reinstates, resulting in an increase in fluorescence intensity. As a rare example amongst indocyanines, one of our new probes was found suitable in STED-based super-resolution imaging...
February 12, 2018: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/29422066/genome-wide-analyses-reveal-a-role-of-polycomb-in-promoting-hypomethylation-of-dna-methylation-valleys
#2
Yuanyuan Li, Hui Zheng, Qiujun Wang, Chen Zhou, Lei Wei, Xuehui Liu, Wenhao Zhang, Yu Zhang, Zhenhai Du, Xiaowo Wang, Wei Xie
BACKGROUND: Previous studies showed that the majority of developmental genes are devoid of DNA methylation at promoters even when they are repressed. Such hypomethylated regions at developmental genes are unusually large and extend well beyond proximal promoters, forming DNA methylation valleys (DMVs) or DNA methylation canyons. However, it remains elusive how most developmental genes can evade DNA methylation regardless of their transcriptional states. RESULTS: We show that DMVs are hypomethylated in development and are highly conserved across vertebrates...
February 8, 2018: Genome Biology
https://www.readbyqxmd.com/read/29405000/site-specific-protein-labeling-with-tetrazine-amino-acids
#3
Robert J Blizzard, True E Gibson, Ryan A Mehl
Genetic code expansion is commonly used to introduce bioorthogonal reactive functional groups onto proteins for labeling. In recent years, the inverse electron demand Diels-Alder reaction between tetrazines and strained trans-cyclooctenes has increased in popularity as a bioorthogonal ligation for protein labeling due to its fast reaction rate and high in vivo stability. We provide methods for the facile synthesis of a tetrazine containing amino acid, Tet-v2.0, and the site-specific incorporation of Tet-v2...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29377916/vaccination-potential-of-b-and-t-epitope-enriched-np-and-m2-against-influenza-a-viruses-from-different-clades-and-hosts
#4
Irina Tutykhina, Ilias Esmagambetov, Alexander Bagaev, Alexey Pichugin, Andrey Lysenko, Dmitry Shcherbinin, Elena Sedova, Denis Logunov, Maxim Shmarov, Ravshan Ataullakhanov, Boris Naroditsky, Alexander Gintsburg
To avoid outbreaks of influenza virus epidemics and pandemics among human populations, modern medicine requires the development of new universal vaccines that are able to provide protection from a wide range of influenza A virus strains. In the course of development of a universal vaccine, it is necessary to consider that immunity must be generated even against viruses from different hosts because new human epidemic virus strains have their origins in viruses of birds and other animals. We have enriched conserved viral proteins-nucleoprotein (NP) and matrix protein 2 (M2)-by B and T-cell epitopes not only human origin but also swine and avian origin...
2018: PloS One
https://www.readbyqxmd.com/read/29374200/interplay-between-tets-and-micrornas-in-the-adult-brain-for-memory-formation
#5
Eloïse A Kremer, Niharika Gaur, Melissa A Lee, Olivia Engmann, Johannes Bohacek, Isabelle M Mansuy
5-hydroxymethylation (5-hmC) is an epigenetic modification on DNA that results from the conversion of 5-methylcytosine by Ten-Eleven Translocation (TET) proteins. 5-hmC is widely present in the brain and is subjected to dynamic regulation during development and upon neuronal activity. It was recently shown to be involved in memory processes but currently, little is known about how it is controlled in the brain during memory formation. Here, we show that Tet3 is selectively up-regulated by activity in hippocampal neurons in vitro, and after formation of fear memory in the hippocampus...
January 26, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29367928/dynamics-of-transgenic-enterobacter-cloacae-expressing-green-fluorescent-protein-defensin-gfp-d-in-anopheles-stephensi-under-laboratory-condition
#6
Hossein Dehghan, Mohammad Ali Oshaghi, Seyed Hassan Moosa-Kazemi, Bagher Yakhchali, Hassan Vatandoost, Naseh Maleki-Ravasan, Yavar Rassi, Habib Mohammadzadeh, Mohammad Reza Abai, Fatemeh Mohtarami
Background: Enterobacter cloacae bacterium is a known symbiont of the most Anopheles gut microflora and nominated as a good candidate for paratransgenic control of malaria. However, the population dynamics of this bacterium within An. stephensi and its introduction methods to the mosquitoes have not yet been explored. Methods: Enterobacter cloacae subsp. dissolvens expressing green fluorescent protein and defensin (GFP-D) was used to study transstadial transmission and the course of time, larval habitat, sugar, and blood meal on dynamics of the bacterium in the mosquito life stages in the laboratory condition...
December 2017: Journal of Arthropod-borne Diseases
https://www.readbyqxmd.com/read/29364877/tet2-promotes-pathogen-infection-induced-myelopoiesis-through-mrna-oxidation
#7
Qicong Shen, Qian Zhang, Yang Shi, Qingzhu Shi, Yanyan Jiang, Yan Gu, Zhiqing Li, Xia Li, Kai Zhao, Chunmei Wang, Nan Li, Xuetao Cao
Varieties of RNA modification form the epitranscriptome for post-transcriptional regulation. 5-Methylcytosine (5-mC) is a sparse RNA modification in messenger RNA (mRNA) under physiological conditions. The function of RNA 5-hydroxymethylcytosine (5-hmC) oxidized by ten-eleven translocation (Tet) proteins in Drosophila has been revealed more recently. However, the turnover and function of 5-mC in mammalian mRNA have been largely unknown. Tet2 suppresses myeloid malignancies mostly in an enzymatic activity-dependent manner, and is important in resolving inflammatory response in an enzymatic activity-independent way...
February 1, 2018: Nature
https://www.readbyqxmd.com/read/29353442/flexible-pseudotyping-of-retrovirus-using-recombinase-mediated-cassette-exchange
#8
Hugo R Soares, Ana I Almeida, Hélio A Tomás, Paula M Alves, Ana S Coroadinha
OBJECTIVE: Develop an engineered cell line containing two flexible gene expression systems enabling the continuous production of tailor-made recombinant gammaretrovirus with predictable productivities through targeted integration. RESULTS: Dual-FLEX cells (dFLEX) contain two independent recombinase-mediated cassette exchange (RMCE) systems which confer flexibility to the expression of different transgene and envelope combinations. The flexible envelope expression in dFLEX cells was validated by pseudotyping retrovirus particles with three different viral envelope proteins-GaLV, 4070A and VSV-G...
January 20, 2018: Biotechnology Letters
https://www.readbyqxmd.com/read/29345269/tributyltin-induces-epigenetic-changes-and-decreases-the-expression-of-nuclear-respiratory-factor-1
#9
Saki Hanaoka, Keishi Ishida, Saki Tanaka, Shuichiro Sakamoto, Katsuhiro Okuda, Seigo Sanoh, Shigeru Ohta, Yaichiro Kotake
Tributyltin (TBT), a common organotin environmental pollutant, has been widely used as a component of marine antifouling paints. We previously reported that exposure to TBT inhibits the expression and DNA binding of nuclear respiratory factor-1 (NRF-1) and causes neurotoxicity. In the present study, we focused on the epigenetic effects of TBT and investigated whether TBT decreases NRF-1 expression via epigenetic modifications in SH-SY5Y human neuroblastoma cells. First, we found that exposure to 300 nM TBT decreases NRF-1 expression...
January 18, 2018: Metallomics: Integrated Biometal Science
https://www.readbyqxmd.com/read/29331106/kbtbd2-inhibits-the-cytotoxic-activity-of-immortalized-nk-cells-through-down-regulating-mtor-signaling-in-a-mouse-hepatocellular-carcinoma-model
#10
Kai Dai, Yabing Huang, Zubing Chen, Xiaomei Sun, Lihua Yang, Yingan Jiang
Natural killer cell (NK cell)-based immunotherapy is a promising therapeutic strategy for hepatocellular carcinoma (HCC). However, the molecular mechanisms underlying the regulation of NK cell function in the tumor sites are not completely elucidated. In this study, we identified the enhanced expression of kelch repeat and BTB (POZ) domain containing 2 (Kbtbd2) in intratumoral NK cells in a mouse HCC implantation model as a negative regulator of NK cells. To investigate this interaction,, we used a Tet-on inducible expression system to control Kbtbd2 expression in an immortalized mouse NK cell line KIL C...
January 13, 2018: European Journal of Immunology
https://www.readbyqxmd.com/read/29324752/tet-protein-function-during-drosophila-development
#11
Fei Wang, Svetlana Minakhina, Hiep Tran, Neha Changela, Joseph Kramer, Ruth Steward
The TET (Ten-eleven translocation) 1, 2 and 3 proteins have been shown to function as DNA hydroxymethylases in vertebrates and their requirements have been documented extensively. Recently, the Tet proteins have been shown to also hydroxylate 5-methylcytosine in RNA. 5-hydroxymethylcytosine (5hmrC) is enriched in messenger RNA but the function of this modification has yet to be elucidated. Because Cytosine methylation in DNA is barely detectable in Drosophila, it serves as an ideal model to study the biological function of 5hmrC...
2018: PloS One
https://www.readbyqxmd.com/read/29314736/development-of-a-chlamydia-suis-specific-antibody-enzyme-linked-immunosorbent-assay-based-on-the-use-of-a-b-cell-epitope-of-the-polymorphic-membrane-protein-c
#12
K De Puysseleyr, E Kieckens, L De Puysseleyr, H Van den Wyngaert, B Ahmed, S Van Lent, H H Creasy, G S A Myers, D Vanrompay
Chlamydia suis infections lead to economic loss in the pork industry. Chlamydia suis infections could be successfully treated with tetracyclines until the appearance of a tetracycline resistant phenotype, which was acquired via horizontal gene transfer of the tet(C) gene. Given the importance of C. suis as a swine pathogen and as a recently emerged tetracycline resistant pathogen with zoonotic potential, our aim was to develop a sensitive C. suis-specific antibody ELISA based on the polymorphic membrane proteins (Pmps)...
January 4, 2018: Transboundary and Emerging Diseases
https://www.readbyqxmd.com/read/29313467/temporal-expression-of-mutant-tdp-43-correlates-with-early-amyotrophic-lateral-sclerosis-phenotype-and-motor-weakness
#13
Qihua Chen, Jinxia Zhou, Cao Huang, Bo Huang, Fangfang Bi, Hongxia Zhou, Bo Xiao
BACKGROUND: Mutant transactive response DNA-binding protein (TDP-43) is closely correlated to inherited form of amyotrophic lateral sclerosis (ALS). TDP-43 transgenic rats can reproduce the core phenotype of ALS and constitutive expression of TDP-43 caused postnatal death. OBJECTIVE: To understand whether neurologic deficiency caused by mutant TDP-43 is dependent on its temporal expression. METHOD: Transgenic rats were established that express mutant human TDP-43 (M337V substitution) in neurons, then a Tet-off system was used to regulate its expression...
January 9, 2018: Current Neurovascular Research
https://www.readbyqxmd.com/read/29290818/methylation-associated-silencing-of-mir-193a-3p-promotes-ovarian-cancer-aggressiveness-by-targeting-grb7-and-mapk-erk-pathways
#14
Kangmei Chen, Michelle Xin Liu, Celia Sze-Ling Mak, Mingo Ming-Ho Yung, Thomas Ho-Yin Leung, Dakang Xu, Siew-Fei Ngu, Karen Kar-Loen Chan, Huijuan Yang, Hextan Yuen-Sheung Ngan, David Wai Chan
Human growth factor receptor-bound protein-7 (GRB7) is a pivotal mediator involved in receptor tyrosine kinase signaling and governing diverse cellular processes. Aberrant upregulation of GRB7 is frequently associated with the progression of human cancers. However, the molecular mechanisms leading to the upregulation of GRB7 remain largely unknown. Here, we propose that the epigenetic modification of GRB7 at the post-transcriptional level may be a crucial factor leading to GRB7 upregulation in ovarian cancers...
2018: Theranostics
https://www.readbyqxmd.com/read/29286410/an-engineered-split-tet2-enzyme-for-chemical-inducible-dna-hydroxymethylation-and-epigenetic-remodeling
#15
Minjung Lee, Yubin Zhou, Yun Huang
DNA methylation is a stable and heritable epigenetic modification in the mammalian genome and is involved in regulating gene expression to control cellular functions. The reversal of DNA methylation, or DNA demethylation, is mediated by the ten-eleven translocation (TET) protein family of dioxygenases. Although it has been widely reported that aberrant DNA methylation and demethylation are associated with developmental defects and cancer, how these epigenetic changes directly contribute to the subsequent alteration in gene expression or disease progression remains unclear, largely owing to the lack of reliable tools to accurately add or remove DNA modifications in the genome at defined temporal and spatial resolution...
December 18, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29286360/conditional-knockdown-of-gene-expression-in-cancer-cell-lines-to-study-the-recruitment-of-monocytes-macrophages-to-the-tumor-microenvironment
#16
Marta H Kubala, Yves A DeClerck
siRNA and shRNA-mediated knock down (KD) methods of regulating gene expression are invaluable tools for understanding gene and protein function. However, in the case that the KD of the protein of interest has a lethal effect on cells or the anticipated effect of the KD is time-dependent, unconditional KD methods are not appropriate. Conditional systems are more suitable in these cases and have been the subject of much interest. These include Ecdysone-inducible overexpression systems, Cytochrome P-450 induction system1, and the tetracycline regulated gene expression systems...
November 23, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29285192/ten-eleven-translocation-1-dysfunction-reduces-5-hydroxymethylcytosine-expression-levels-in-gastric-cancer-cells
#17
Kuo-Chiang Wang, Chi-Hsiang Kang, Chung-Yu Tsai, Nan-Hua Chou, Ya-Ting Tu, Guan-Cheng Li, Hing-Chung Lam, Shiuh-Inn Liu, Po-Min Chang, Yan-Hwai Lin, Kuo-Wang Tsai
A sixth base, 5-hydroxymethylcytosine (5hmC), is formed by the oxidation of 5-methylcytosine (5mC) via the catalysis of the ten-eleven translocation (TET) protein family in cells. Expression levels of 5hmC are frequently depleted during carcinogenesis. However, the detailed mechanisms underlying the depletion of 5hmC expression in gastric cancer cells remains unclear, and further research is required. The present study examined the expression levels of 5mC and 5hmC and the expression levels of TET1 and TET2 in gastric cancer tissues using immunohistochemistry...
January 2018: Oncology Letters
https://www.readbyqxmd.com/read/29282319/expression-of-an-active-g%C3%AE-s-mutant-in-skeletal-stem-cells-is-sufficient-and-necessary-for-fibrous-dysplasia-initiation-and-maintenance
#18
Xuefeng Zhao, Peng Deng, Ramiro Iglesias-Bartolome, Panomwat Amornphimoltham, Dana J Steffen, Yunyun Jin, Alfredo A Molinolo, Luis Fernandez de Castro, Diana Ovejero, Quan Yuan, Qianming Chen, Xianglong Han, Ding Bai, Susan S Taylor, Yingzi Yang, Michael T Collins, J Silvio Gutkind
Fibrous dysplasia (FD) is a disease caused by postzygotic activating mutations of GNAS (R201C and R201H) that encode the α-subunit of the Gs stimulatory protein. FD is characterized by the development of areas of abnormal fibroosseous tissue in the bones, resulting in skeletal deformities, fractures, and pain. Despite the well-defined genetic alterations underlying FD, whether GNAS activation is sufficient for FD initiation and the molecular and cellular consequences of GNAS mutations remains largely unresolved, and there are no currently available targeted therapeutic options for FD...
December 27, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29276783/a-method-for-the-efficient-and-selective-identification-of-5-hydroxymethyluracil-in-genomic-dna
#19
Whitney Bullard, Rudo Kieft, Robert Sabatini
Recently, 5-hydroxymethyluracil (5hmU) was identified in mammalian genomic DNA as an oxidative product of thymine by the ten-eleven translocation (TET) proteins. While the biological role of this modification remains unclear, identifying its genomic location will assist in elucidating function. Here we present a rapid and robust method to selectively tag and enrich genomic regions containing 5hmU. This method involves the selective glucosylation of 5hmU residues by the base J glucosyltransferase from trypanosomes creating glucosylhydroxymethyluracil (base J)...
January 2017: Biology Methods and Protocols
https://www.readbyqxmd.com/read/29273640/using-cellular-fitness-to-map-the-structure-and-function-of-a-major-facilitator-superfamily-effluxer
#20
Anisha M Perez, Marcella M Gomez, Prashant Kalvapalle, Erin O'Brien-Gilbert, Matthew R Bennett, Yousif Shamoo
The major facilitator superfamily (MFS) effluxers are prominent mediators of antimicrobial resistance. The biochemical characterization of MFS proteins is hindered by their complex membrane environment that makes in vitro biochemical analysis challenging. Since the physicochemical properties of proteins drive the fitness of an organism, we posed the question of whether we could reverse that relationship and derive meaningful biochemical parameters for a single protein simply from fitness changes it confers under varying strengths of selection...
December 22, 2017: Molecular Systems Biology
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