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https://www.readbyqxmd.com/read/28723582/overexpression-and-purification-of-dicer-and-accessory-proteins-for-biochemical-and-structural-studies
#1
Niladri K Sinha, Brenda L Bass
The Dicer family of ribonucleases plays a key role in small RNA-based regulatory pathways by generating short dsRNA fragments that modulate expression of endogenous genes, or protect the host from invasive nucleic acids. Beginning with its initial discovery, biochemical characterization of Dicer has provided insight about its catalytic properties. However, a comprehensive understanding of how Dicer's domains contribute to substrate-specific recognition and catalysis is lacking. One reason for this void is the lack of high-resolution structural information for a metazoan Dicer in the apo- or substrate-bound state...
July 16, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/28717424/low-active-loading-of-cargo-into-engineered-extracellular-vesicles-results-in-inefficient-mirna-mimic-delivery
#2
Dhruvitkumar S Sutaria, Jinmai Jiang, Ola A Elgamal, Steven M Pomeroy, Mohamed Badawi, Xiaohua Zhu, Ryan Pavlovicz, Ana Clara P Azevedo-Pouly, Jeffrey Chalmers, Chenglong Li, Mitch A Phelps, Thomas D Schmittgen
Extracellular vesicles (EVs) hold great potential as novel systems for nucleic acid delivery due to their natural composition. Our goal was to load EVs with microRNA that are synthesized by the cells that produce the EVs. HEK293T cells were engineered to produce EVs expressing a lysosomal associated membrane, Lamp2a fusion protein. The gene encoding pre-miR-199a was inserted into an artificial intron of the Lamp2a fusion protein. The TAT peptide/HIV-1 transactivation response (TAR) RNA interacting peptide was exploited to enhance the EV loading of the pre-miR-199a containing a modified TAR RNA loop...
2017: Journal of Extracellular Vesicles
https://www.readbyqxmd.com/read/28716689/brg1-and-smarcal1-transcriptionally-co-regulate-drosha-dgcr8-and-dicer-in-response-to-doxorubicin-induced-dna-damage
#3
Ketki Patne, Radhakrishnan Rakesh, Vijendra Arya, Upasana Bedi Chanana, Ramesh Sethy, Pynskhem Bok Swer, Rohini Muthuswami
Recent investigations have emphasized the role of miRNA biogenesis proteins in the synthesis of non-coding RNA when double-strand DNA breaks are induced by ionizing radiations. However, the role of these ncRNA and their regulation in response to doxorubicin-induced DNA damage is not known. In this paper, BRG1 and SMARCAL1, members of the ATP-dependent chromatin remodelling family, are shown to co-regulate the transcription of DROSHA, DGCR8, and DICER in response to double-strand DNA breaks induced by doxorubicin...
July 14, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28704808/enhanced-reactive-oxygen-species-production-acidic-cytosolic-ph-and-upregulated-na-h-exchanger-nhe-in-dicer-deficient-cd4-t-cells
#4
Yogesh Singh, Yuetao Zhou, Shaqiu Zhang, Khalid N M Abdelazeem, Bernat Elvira, Madhuri S Salker, Florian Lang
BACKGROUND: MicroRNAs (miRNAs) negatively regulate gene expression at a post-transcriptional level. Dicer, a cytoplasmic RNase III enzyme, is required for the maturation of miRNAs from precursor miRNAs. Dicer, therefore, is a critical enzyme involved in the biogenesis and processing of miRNAs. Several biological processes are controlled by miRNAs, including the regulation of T cell development and function. T cells generate reactive oxygen species (ROS) with parallel H+ extrusion accomplished by the Na+/H+-exchanger 1 (NHE1)...
July 14, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28700949/two-sets-of-piwi-proteins-are-involved-in-distinct-srna-pathways-leading-to-elimination-of-germline-specific-dna
#5
Dominique I Furrer, Estienne C Swart, Matthias F Kraft, Pamela Y Sandoval, Mariusz Nowacki
Piwi proteins and piRNAs protect eukaryotic germlines against the spread of transposons. During development in the ciliate Paramecium, two Piwi-dependent sRNA classes are involved in the elimination of transposons and transposon-derived DNA: scan RNAs (scnRNAs), associated with Ptiwi01 and Ptiwi09, and iesRNAs, whose binding partners we now identify as Ptiwi10 and Ptiwi11. scnRNAs derive from the maternal genome and initiate DNA elimination during development, whereas iesRNAs continue DNA targeting until the removal process is complete...
July 11, 2017: Cell Reports
https://www.readbyqxmd.com/read/28696509/an-l-rna-aptamer-with-expanded-chemical-functionality-that-inhibits-microrna-biogenesis
#6
Adam Michael Kabza, Jonathan Thomas Sczepanski
In order to facilitate isolation of L-aptamers with novel RNA-binding properties, we have employed a cationic nucleotide, 5-aminoallyl-uridine, during the mirror image in vitro selection process. In this effort, we identified a modified L-RNA aptamer (MLRA) capable of binding oncogenic pre-miR-19a with exceptional affinity, and we show that the cationic modification is absolutely critical for binding. Furthermore, formation of the MLRA-pre-miR-19a complex inhibits Dicer-mediated cleavage of the pre-miR, thus blocking formation of the mature functional microRNA...
July 11, 2017: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/28695996/downregulation-of-genes-coding-for-core-rnai-components-and-disease-resistance-proteins-via-corresponding-micrornas-might-be-correlated-with-successful-smv-infection-in-soybean
#7
Duran Bao, Oyunchuluun Ganbaatar, Xiuqi Cui, Ruonan Yu, Wenhua Bao, Bryce W Falk, Hada Wuriyanghan
Plants protect themselves from virus infections by several different defense mechanisms. RNA interference (RNAi) is one prominent antiviral mechanism, which requires the participation of AGO (Argonaute) and Dicer/DCL (Dicer-like) proteins. Effector-triggered immunity (ETI) is an antiviral mechanism mediated by resistance (R) genes, most of which encode NBS-LRR family proteins. MicroRNAs (miRNAs) play important regulatory roles in plants, including regulation of host defenses. SMV (Soybean mosaic virus) is the most common virus in soybean and here we identified dozens of SMV-responsive miRNAs by microarray analysis in a SMV-susceptible soybean line...
July 11, 2017: Molecular Plant Pathology
https://www.readbyqxmd.com/read/28690605/dicer-like-genes-are-required-for-h2o2-and-kcl-stress-responses-pathogenicity-and-small-rna-generation-in-valsa-mali
#8
Hao Feng, Ming Xu, Yangyang Liu, Ruqing Dong, Xiaoning Gao, Lili Huang
Valsa mali (V. mali) is the causative agent of apple tree Valsa canker, which heavily damages the production of apples in China. However, the biological roles of the RNA interfering (RNAi) pathway in the pathogenicity of V. mali remain unknown. Dicer-like proteins (DCLs) are important components that control the initiation of the RNAi pathway. In this study, VmDCL1 and VmDCL2 were isolated and functionally characterized in V. mali. VmDCL1 and VmDCL2 are orthologous in evolution to the DCLs in Cryphonectria parasitica...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28686847/development-and-implementation-of-an-hts-compatible-assay-for-the-discovery-of-selective-small-molecule-ligands-for-pre-micrornas
#9
Daniel A Lorenz, Steve Vander Roest, Martha J Larsen, Amanda L Garner
microRNAs (miRNAs) are small gene regulatory RNAs, and their expression has been found to be dysregulated in a number of human diseases. To facilitate the discovery of small molecules capable of selectively modulating the activity of a specific miRNA, we have utilized new high-throughput screening technology targeting Dicer-mediated pre-miRNA maturation. Pilot screening of ~50,000 small molecules and ~33,000 natural product extract libraries against pre-miR-21 processing indicated the potential of our assay for this goal, yielding a campaign Z' factor of 0...
July 1, 2017: SLAS Discovery
https://www.readbyqxmd.com/read/28674223/expression-analysis-of-argonaute-dicer-like-and-rna-dependent-rna-polymerase-genes-in-cucumber-cucumis-sativus-l-in-response-to-abiotic-stress
#10
Defang Gan, Mengdan Zhan, Feng Yang, Qiqi Zhang, Keling Hu, Wenjuan Xu, Qinghui Lu, Ling Zhang, Dandi Liang
Posttranscriptional control of gene expression can be achieved through RNA interference when the activities of Dicer-like (DCL), argonaute (AGO) and RNA-dependent RNA polymerase (RDR) proteins are significant. In this study, we analysed the expression of seven AGO, five DCL and eight RDR genes in cucumber under cold, heat, hormone, salinity and dehydration treatments using quantitative reverse-transcription PCR (qRT-PCR). All CsAGO, CsDCL and CsRDR genes were differentially expressed under abiotic stress treatment...
June 2017: Journal of Genetics
https://www.readbyqxmd.com/read/28670496/resistance-to-cancer-chemotherapeutic-drugs-is-determined-by-pivotal-microrna-regulators
#11
Marta Geretto, Alessandra Pulliero, Camillo Rosano, Dinara Zhabayeva, Rakhmet Bersimbaev, Alberto Izzotti
Chemo-resistance, which is the main obstacle in cancer therapy, is caused by the onset of drug-resistant cells in the heterogeneous cell population in cancer tissues. MicroRNAs regulate gene expression at the post-transcriptional level, and they are involved in many different biological processes, including cell proliferation, differentiation, metabolism, stress response, and apoptosis. The aberrant expression of microRNAs plays a major pathogenic role from the early stages of the carcinogenesis process. Recently, microRNAs have been reported to play an important role in inducing resistance to anti-cancer drugs...
2017: American Journal of Cancer Research
https://www.readbyqxmd.com/read/28665784/the-evidence-for-a-microrna-product-of-human-drosha-gene
#12
Peter Mechtler, Sydney Johnson, Hannah Slabodkin, Amir B Cohanim, Leonid Brodsky, Eugene S Kandel
MicroRNAs are short RNA molecules that regulate function and stability of a large subset of eukaryotic mRNAs. In the main pathway of microRNA biogenesis, a short "hairpin" is excised from a primary transcript by ribonuclease DROSHA, followed by additional nucleolytic processing by DICER and inclusion of the mature microRNA into the RNA-induced silencing complex. We report that a microRNA-like molecule is encoded by human DROSHA gene within a predicted stem-loop element of the respective transcript. This putative mature microRNA is complementary to DROSHA transcript variant 1 and can attenuate expression of the corresponding protein...
June 30, 2017: RNA Biology
https://www.readbyqxmd.com/read/28665774/new-drb-complexes-for-new-drb-functions-in-plants
#13
Thomas Montavon, Yerim Kwon, Aude Zimmermann, Fabrice Michel, Patrice Dunoyer
Double-stranded RNA binding (DRB) proteins are generally considered as promoting cofactors of Dicer or Dicer-like (DCL) proteins that ensure efficient and precise production of small RNAs, the sequence-specificity guide of RNA silencing processes in both plants and animals. However, the characterization of a new clade of DRB proteins in Arabidopsis has recently challenged this view by showing that DRBs can also act as potent inhibitors of DCL processing. This is achieved through sequestration of a specific class of small RNA precursors, the endogenous inverted-repeat (endoIR) dsRNAs, thereby selectively preventing production of their associated small RNAs, the endoIR-siRNAs...
June 30, 2017: RNA Biology
https://www.readbyqxmd.com/read/28650464/%C3%AE-catenin-downregulates-dicer-to-promote-ovarian-cancer-metastasis
#14
S K Y To, A S C Mak, Y M Eva Fung, C-M Che, S-S Li, W Deng, B Ru, J Zhang, A S T Wong
Ovarian cancer is a nearly uniform lethal disease and its highly aggressive metastatic phenotype portends a poor prognosis. Lack of a well-controlled, relevant experimental model has been a major obstacle to identifying key molecules causing metastasis. Here we describe the creation of a new isogenic model of spontaneous human ovarian cancer metastasis exhibiting opposite phenotypes-highly metastatic (HM) and non-metastatic (NM)-both in vitro and in vivo. HM was unique in its ability to metastasize consistently to the peritoneum, mimicking the major dissemination route of human ovarian cancer...
June 26, 2017: Oncogene
https://www.readbyqxmd.com/read/28648842/pnrc2-regulates-3-utr-mediated-decay-of-segmentation-clock-associated-transcripts-during-zebrafish-segmentation
#15
Thomas L Gallagher, Kiel T Tietz, Zachary T Morrow, Jasmine M McCammon, Michael L Goldrich, Nicolas L Derr, Sharon L Amacher
Vertebrate segmentation is controlled by the segmentation clock, a molecular oscillator that regulates gene expression and cycles rapidly. The expression of many genes oscillates during segmentation, including hairy/Enhancer of split-related (her or Hes) genes, which encode transcriptional repressors that auto-inhibit their own expression, and deltaC (dlc), which encodes a Notch ligand. We previously identified the tortuga (tor) locus in a zebrafish forward genetic screen for genes involved in cyclic transcript regulation and showed that cyclic transcripts accumulate post-splicing in tor mutants...
June 23, 2017: Developmental Biology
https://www.readbyqxmd.com/read/28643928/calvarial-bone-development-and-suture-closure-in-dicer-deficient-mice
#16
P Atsawasuwan, M Ouibaidin, B Dalal, H Khan, A Mohammed
OBJECTIVE: To evaluate whether lack of Dicer during calvaria development would lead to dysmorphology of calvaria and suture closure in mice. MATERIALS AND METHODS: A conditional Dicer deficient under Osx promoter mouse was employed in this study. The 4- and 10-week-old conditional Dicer-deficient mice control littermates and Osx-cre transgenic mice were studied for calvarial bone morphology and suture closure. Dry skull, microcomputed tomography (μCT), histological and gene expression studies were investigated to evaluate the effect of Dicer deficiency on calvarial bone morphology and their related genes during calvaria development...
June 2017: Orthodontics & Craniofacial Research
https://www.readbyqxmd.com/read/28642363/nuclear-phosphorylated-dicer-processes-double-stranded-rna-in-response-to-dna-damage
#17
Kaspar Burger, Margarita Schlackow, Martin Potts, Svenja Hester, Shabaz Mohammed, Monika Gullerova
The endoribonuclease Dicer is a key component of the human RNA interference pathway and is known for its role in cytoplasmic microRNA production. Recent findings suggest that noncanonical Dicer generates small noncoding RNA to mediate the DNA damage response (DDR). Here, we show that human Dicer is phosphorylated in the platform-Piwi/Argonaute/Zwille-connector helix cassette (S1016) upon induction of DNA damage. Phosphorylated Dicer (p-Dicer) accumulates in the nucleus and is recruited to DNA double-strand breaks...
June 22, 2017: Journal of Cell Biology
https://www.readbyqxmd.com/read/28641547/recent-advances-on-the-role-of-micrornas-in-both-insulin-resistance-and-cancer
#18
Adele Vivacqua, Paola De Marco, Antonino Belfiore, Marcello Maggiolini
BACKGROUND: Insulin resistance is a pathological condition characterized by the failure of target cells to uptake and metabolizes glucose in response to insulin. In particular, the elevated concentrations of glucose, insulin and free insulin growth factor-1, which result from insulin resistance, may generate a pro-inflammatory and pro-tumorigenic state. These alterations may underlie the increased risk to develop various types of cancer as well as the worse cancer prognosis observed in obese and diabetic patients...
June 22, 2017: Current Pharmaceutical Design
https://www.readbyqxmd.com/read/28637695/corrigendum-dicer-loss-and-recovery-induce-an-oncogenic-switch-driven-by-transcriptional-activation-of-the-oncofetal-imp1-3-family
#19
Courtney K JnBaptiste, Allan M Gurtan, Kevin K Thai, Victoria Lu, Arjun Bhutkar, Mei-Ju Su, Asaf Rotem, Tyler Jacks, Phillip A Sharp
No abstract text is available yet for this article.
May 15, 2017: Genes & Development
https://www.readbyqxmd.com/read/28636969/human-virus-derived-small-rnas-can-confer-antiviral-immunity-in-mammals
#20
Yang Qiu, Yanpeng Xu, Yao Zhang, Hui Zhou, Yong-Qiang Deng, Xiao-Feng Li, Meng Miao, Qiang Zhang, Bo Zhong, Yuanyang Hu, Fu-Chun Zhang, Ligang Wu, Cheng-Feng Qin, Xi Zhou
RNA interference (RNAi) functions as a potent antiviral immunity in plants and invertebrates; however, whether RNAi plays antiviral roles in mammals remains unclear. Here, using human enterovirus 71 (HEV71) as a model, we showed HEV71 3A protein as an authentic viral suppressor of RNAi during viral infection. When the 3A-mediated RNAi suppression was impaired, the mutant HEV71 readily triggered the production of abundant HEV71-derived small RNAs with canonical siRNA properties in cells and mice. These virus-derived siRNAs were produced from viral dsRNA replicative intermediates in a Dicer-dependent manner and loaded into AGO, and they were fully active in degrading cognate viral RNAs...
June 20, 2017: Immunity
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