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https://www.readbyqxmd.com/read/28938749/the-testicular-transcriptome-associated-with-spermatogonia-differentiation-initiated-by-gonadotrophin-stimulation-in-the-juvenile-rhesus-monkey-macaca-mulatta
#1
Suresh Ramaswamy, William H Walker, Paula Aliberti, Rahil Sethi, Gary R Marshall, Alyxzandria Smith, Seyedmehdi Nourashrafeddin, Alicia Belgorosky, Uma R Chandran, Mark P Hedger, Tony M Plant
STUDY QUESTION: What is the genetic landscape within the testis of the juvenile rhesus monkey (Macaca mulatta) that underlies the decision of undifferentiated spermatogonia to commit to a pathway of differentiation when puberty is induced prematurely by exogenous LH and FSH stimulation? SUMMARY ANSWER: Forty-eight hours of gonadotrophin stimulation of the juvenile monkey testis resulted in the appearance of differentiating B spermatogonia and the emergence of 1362 up-regulated and 225 down-regulated testicular mRNAs encoding a complex network of proteins ranging from enzymes regulating Leydig cell steroidogenesis to membrane receptors, and from juxtacrine and paracrine factors to transcriptional factors governing spermatogonial stem cell fate...
October 1, 2017: Human Reproduction
https://www.readbyqxmd.com/read/28938739/sequence-variants-of-khdrbs1-as-high-penetrance-susceptibility-risks-for-primary-ovarian-insufficiency-by-mis-regulating-mrna-alternative-splicing
#2
Binbin Wang, Lin Li, Ying Zhu, Wei Zhang, Xi Wang, Beili Chen, Tengyan Li, Hong Pan, Jing Wang, Kehkooi Kee, Yunxia Cao
STUDY QUESTION: Does a novel heterozygous KHDRBS1 variant, identified using whole-exome sequencing (WES) in two patients with primary ovarian insufficiency (POI) in a pedigree, cause defects in mRNA alternative splicing? SUMMARY ANSWER: The heterozygous variant of KHDRBS1 was confirmed to cause defects in alternative splicing of many genes involved in DNA replication and repair. WHAT IS KNOWN ALREADY: Studies in mice revealed that Khdrbs1 deficient females are subfertile, which manifests as delayed sexual maturity and significantly reduced numbers of secondary and pre-antral follicles...
October 1, 2017: Human Reproduction
https://www.readbyqxmd.com/read/28938481/vesicular-glutamate-transporter-1-knockdown-in-infralimbic-prefrontal-cortex-augments-neuroendocrine-responses-to-chronic-stress-in-male-rats
#3
Brent Myers, Jessica M McKlveen, Rachel Morano, Yvonne M Ulrich-Lai, Matia B Solomon, Steven P Wilson, James P Herman
Chronic stress-associated pathologies frequently associate with alterations in the structure and activity of medial prefrontal cortex (mPFC). However, the influence of infralimbic cortex (IL) projection neurons on HPA axis activity is unknown, as is the involvement of these cells in chronic stress-induced endocrine alterations. In the current study, a lentiviral-packaged vector coding for a small interfering RNA (siRNA) targeting vesicular glutamate transporter 1 (vGluT1) mRNA was microinjected into the IL of male rats...
July 25, 2017: Endocrinology
https://www.readbyqxmd.com/read/28938099/plasmid-dna-launches-live-attenuated-japanese-encephalitis-virus-and-elicits-virus-neutralizing-antibodies-in-balb-c-mice
#4
Brian Nickols, Irina Tretyakova, Alexander Tibbens, Elena Klyushnenkova, Peter Pushko
We describe novel plasmid DNA that encodes the full-length Japanese encephalitis virus (JEV) genomic cDNA and launches live-attenuated JEV vaccine in vitro and in vivo. The synthetic cDNA based on the sequence of JEV SA14-14-2 live-attenuated virus was placed under transcriptional control of the cytomegalovirus major immediate-early promoter. The stability and yields of the plasmid in E. coli were optimized by inserting three synthetic introns that disrupted JEV cDNA in the structural and nonstructural genes...
September 19, 2017: Virology
https://www.readbyqxmd.com/read/28938092/ctcf-mediated-enhancer-promoter-interaction-is-a-critical-regulator-of-cell-to-cell-variation-of-gene-expression
#5
Gang Ren, Wenfei Jin, Kairong Cui, Joseph Rodrigez, Gangqing Hu, Zhiying Zhang, Daniel R Larson, Keji Zhao
Recent studies indicate that even a homogeneous population of cells display heterogeneity in gene expression and response to environmental stimuli. Although promoter structure critically influences the cell-to-cell variation of gene expression in bacteria and lower eukaryotes, it remains unclear what controls the gene expression noise in mammals. Here we report that CTCF decreases cell-to-cell variation of expression by stabilizing enhancer-promoter interaction. We show that CTCF binding sites are interwoven with enhancers within topologically associated domains (TADs) and a positive correlation is found between CTCF binding and the activity of the associated enhancers...
September 21, 2017: Molecular Cell
https://www.readbyqxmd.com/read/28937999/structures-of-foot-and-mouth-disease-virus-pentamers-insight-into-capsid-dissociation-and-unexpected-pentamer-reassociation
#6
Nayab Malik, Abhay Kotecha, Sarah Gold, Amin Asfor, Jingshan Ren, Juha T Huiskonen, Tobias J Tuthill, Elizabeth E Fry, David I Stuart
Foot-and-mouth disease virus (FMDV) belongs to the aphthovirus genus of the Picornaviridae, a family of small, icosahedral, non-enveloped, single-stranded RNA viruses. It is a highly infectious pathogen and is one of the biggest hindrances to the international trade of animals and animal products. FMDV capsids (which are unstable below pH6.5) release their genome into the host cell from an acidic compartment, such as that of an endosome, and in the process dissociate into pentamers. Whilst other members of the family (enteroviruses) have been visualized to form an expanded intermediate capsid with holes from which inner capsid proteins (VP4), N-termini (VP1) and RNA can be released, there has been no visualization of any such state for an aphthovirus, instead the capsid appears to simply dissociate into pentamers...
September 22, 2017: PLoS Pathogens
https://www.readbyqxmd.com/read/28937776/4-c-methoxy-2-deoxy-2-fluoro-modified-ribonucleotides-improve-metabolic-stability-and-elicit-efficient-rnai-mediated-gene-silencing
#7
Elise Malek-Adamian, Dale Guenther, Shigeo Matsuda, Saúl Martínez-Montero, Ivan Zlatev, Joel Harp, Mihai Burai Patrascu, Donald J Foster, Johans J Fakhoury, Lydia Perkins, Nicolas Moitessier, Rajar Manoharan, Nate Taneja, Anna Bisbe, Klaus Charisse, Martin A Maier, Kallanthottathil G Rajeev, Martin Egli, Muthiah Manoharan, Masad J Damha
We designed novel 4'-modified 2'-deoxy-2'-fluorouridine (2'-F U) analogues with the aim to improve nuclease resistance and potency of therapeutic siRNAs by introducing 4'-C-methoxy (4'-OMe) as the alpha (C4'α) or beta (C4'β) epimers. The C4'α epimer was synthesized by a stereoselective route in six steps; however, both α and β epimers could be obtained by a non-stereoselective approach starting from 2'-F U. (1)H-NMR analysis and computational investigation of the α-epimer revealed that the 4'-OMe imparts a conformational bias towards the North-East sugar pucker, due to intramolecular hydrogen bonding and hyperconjugation effects...
September 22, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28936836/-molecular-cloning-and-characterization-of-two-kind-of-heat-shock-protein-gene-from-polyporus-umbellatus
#8
Meng-Meng Liu, Yong-Mei Xing, Shun-Xing Guo
With RT-PCR approaches, the full-length cDNA of two heat shock protein genes were cloned from total RNA of the Polyporus umbellatus sclerotium. The full open reading frame cDNA sequence of the Hsp90 was 2 091 bp, encoding 696 amino acid residues with a predicted molecular mass of 78.9 kDa. The full open reading frame cDNA sequence of the Hsp70 was 1 944 bp, encoding 647 amino acid residues with a predicted molecular mass of 70.5 kDa. The Hsp90 and Hsp70 protein contained the conservative structure domain, respectively...
December 2016: Zhongguo Zhong Yao za Zhi, Zhongguo Zhongyao Zazhi, China Journal of Chinese Materia Medica
https://www.readbyqxmd.com/read/28935965/aromatic-side-chain-conformational-switch-on-the-surface-of-the-rna-recognition-motif-enables-rna-discrimination
#9
Nana Diarra Dit Konté, Miroslav Krepl, Fred F Damberger, Nina Ripin, Olivier Duss, Jiří Šponer, Frédéric H-T Allain
The cyclooxygenase-2 is a pro-inflammatory and cancer marker, whose mRNA stability and translation is regulated by the CUG-binding protein 2 interacting with AU-rich sequences in the 3' untranslated region. Here, we present the solution NMR structure of CUG-binding protein 2 RRM3 in complex with 5'-UUUAA-3' originating from the COX-2 3'-UTR. We show that RRM3 uses the same binding surface and protein moieties to interact with AU- and UG-rich RNA motifs, binding with low and high affinity, respectively. Using NMR spectroscopy, isothermal titration calorimetry and molecular dynamics simulations, we demonstrate that distinct sub-states characterized by different aromatic side-chain conformations at the RNA-binding surface allow for high- or low-affinity binding with functional implications...
September 21, 2017: Nature Communications
https://www.readbyqxmd.com/read/28935900/antibacterial-and-immunomodulatory-activities-of-insect-defensins-dlp2-and-dlp4-against-multidrug-resistant-staphylococcus-aureus
#10
Zhanzhan Li, Ruoyu Mao, Da Teng, Ya Hao, Huixian Chen, Xiumin Wang, Xiao Wang, Na Yang, Jianhua Wang
Methicillin-resistant Staphylococcus aureus (MRSA), are the most frequent cause of sepsis, which urgently demanding new drugs for treating infection. Two homologous insect CSαβ peptides-DLP2 and DLP4 from Hermetia illucens were firstly expressed in Pichia pastoris, with the yields of 873.5 and 801.3 mg/l, respectively. DLP2 and DLP4 displayed potent antimicrobial activity against Gram-positive bacteria especially MRSA and had greater potency, faster killing, and a longer postantibiotic effect than vancomycin...
September 21, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28935721/the-spliceosomal-proteins-ppih-and-prpf4-exhibit-bipartite-binding
#11
Caroline Rajiv, S RaElle Jackson, Simon Cocklin, Elan Z Eisenmesser, Tara Davis
Pre-mRNA splicing is a dynamic, multi-step process that is catalyzed by the RNA: protein complex called the spliceosome. The spliceosome contains a core set of RNAs and proteins that are conserved in all organisms that perform splicing. In higher organisms, PPIH directly interacts with the core protein PRPF4 and both integrate into the pre-catalytic spliceosome as part of the tri-snRNP sub-complex. As a first step to understand the protein interactions that dictate PPIH and PRPF4 function, we expressed and purified soluble forms of each protein and formed a complex between them...
September 21, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28935594/self-replicating-rna-vaccine-functionality-modulated-by-fine-tuning-of-polyplex-delivery-vehicle-structure
#12
Thomas Démoulins, Thomas Ebensen, Kai Schulze, Pavlos C Englezou, Panagiota Milona, Maria Pelliccia, Carlos A Guzmán, Nicolas Ruggli, Kenneth C McCullough
The major limitations with large and complex self-amplifying RNA vaccines (RepRNA) are RNase-sensitivity and inefficient translation in dendritic cells (DCs). Condensing RepRNA with polyethylenimine (PEI) gave positive in vitro readouts, but was largely inferior to virus-like replicon particles (VRP) or direct electroporation. In the present study, we improved such polyplex formulation and determined that fine-tuning of the polyplex structure is essential for ensuring efficacious translation. Thereby, three parameters dominate: (i) PEI molecular weight (MW); (ii) RepRNA:PEI (weight:weight) ratio; and (iii) inclusion of cell penetrating peptides (CPPs)...
September 18, 2017: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/28935369/a-three-dimensional-model-of-rnase-p-in-the-hyperthermophilic-archaeon-pyrococcus-horikoshii-ot3
#13
Xuzhu Gao, Kosuke Oshima, Toshifumi Ueda, Takashi Nakashima, Makoto Kimura
Ribonuclease P (RNase P) is an endoribonuclease involved in maturation of the 5'-end of tRNA. We found previously that RNase P in the hyperthermophilic archaeon Pyrococcus horikoshii OT3 consists of a catalytic RNase P RNA (PhopRNA) and five protein cofactors designated PhoPop5, PhoRpp21, PhoRpp29, PhoRpp30, and PhoRpp38. The crystal structures of the five proteins have been determined, a three-dimensional (3-D) model of PhopRNA has been constructed, and biochemical data, including protein-RNA interaction sites, have become available...
September 18, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28934505/combining-asymmetric-13c-labeling-and-isotopic-filter-edit-noesy-a-novel-strategy-for-rapid-and-logical-rna-resonance-assignment
#14
Regan M LeBlanc, Andrew P Longhini, Stuart F J Le Grice, Bruce A Johnson, Theodore K Dayie
Although ∼98% of the human genomic output is transcribed as non-protein coding RNA, <2% of the protein data bank structures comprise RNA. This huge structural disparity stems from combined difficulties of crystallizing RNA for X-ray crystallography along with extensive chemical shift overlap and broadened linewidths associated with NMR of RNA. While half of the deposited RNA structures in the PDB were solved by NMR methods, the usefulness of NMR is still limited by the high cost of sample preparation and challenges of resonance assignment...
September 19, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28934503/a-novel-requirement-for-drosha-in-maintenance-of-mammalian-cg-methylation
#15
Athanasia Stathopoulou, Jyoti B Chhetri, John C Ambrose, Pierre-Olivier Estève, Lexiang Ji, Hediye Erdjument-Bromage, Guoqiang Zhang, Thomas A Neubert, Sriharsa Pradhan, Javier Herrero, Robert J Schmitz, Steen K T Ooi
In mammals, faithful inheritance of genomic methylation patterns ensures proper gene regulation and cell behaviour, impacting normal development and fertility. Following establishment, genomic methylation patterns are transmitted through S-phase by the maintenance methyltransferase Dnmt1. Using a protein interaction screen, we identify Microprocessor component DROSHA as a novel DNMT1-interactor. Drosha-deficient embryonic stem (ES) cells display genomic hypomethylation that is not accounted for by changes in the levels of DNMT proteins...
September 19, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28934487/supernalign-a-new-tool-for-flexible-superposition-of-homologous-rna-structures-and-inference-of-accurate-structure-based-sequence-alignments
#16
Pawel Piatkowski, Jagoda Jablonska, Adriana Zyla, Dorota Niedzialek, Dorota Matelska, Elzbieta Jankowska, Tomasz Walen, Wayne K Dawson, Janusz M Bujnicki
RNA has been found to play an ever-increasing role in a variety of biological processes. The function of most non-coding RNA molecules depends on their structure. Comparing and classifying macromolecular 3D structures is of crucial importance for structure-based function inference and it is used in the characterization of functional motifs and in structure prediction by comparative modeling. However, compared to the numerous methods for protein structure superposition, there are few tools dedicated to the superimposing of RNA 3D structures...
September 19, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28934484/regulation-of-hur-structure-and-function-by-dihydrotanshinone-i
#17
Preet Lal, Linda Cerofolini, Vito Giuseppe D'Agostino, Chiara Zucal, Carmelo Fuccio, Isabelle Bonomo, Erik Dassi, Stefano Giuntini, Danilo Di Maio, Vikalp Vishwakarma, Ranjan Preet, Sha Neisha Williams, Max S Fairlamb, Rachel Munk, Elin Lehrmann, Kotb Abdelmohsen, Saioa R Elezgarai, Claudio Luchinat, Ettore Novellino, Alessandro Quattrone, Emiliano Biasini, Leonardo Manzoni, Myriam Gorospe, Dan A Dixon, Pierfausto Seneci, Luciana Marinelli, Marco Fragai, Alessandro Provenzani
The Human antigen R protein (HuR) is an RNA-binding protein that recognizes U/AU-rich elements in diverse RNAs through two RNA-recognition motifs, RRM1 and RRM2, and post-transcriptionally regulates the fate of target RNAs. The natural product dihydrotanshinone-I (DHTS) prevents the association of HuR and target RNAs in vitro and in cultured cells by interfering with the binding of HuR to RNA. Here, we report the structural determinants of the interaction between DHTS and HuR and the impact of DHTS on HuR binding to target mRNAs transcriptome-wide...
September 19, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28934478/tuning-rna-folding-and-function-through-rational-design-of-junction-topology
#18
May Daher, Anthony M Mustoe, Alex Morriss-Andrews, Charles L Brooks Iii, Nils G Walter
Structured RNAs such as ribozymes must fold into specific 3D structures to carry out their biological functions. While it is well-known that architectural features such as flexible junctions between helices help guide RNA tertiary folding, the mechanisms through which junctions influence folding remain poorly understood. We combine computational modeling with single molecule Förster resonance energy transfer (smFRET) and catalytic activity measurements to investigate the influence of junction design on the folding and function of the hairpin ribozyme...
September 19, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28934477/a-structural-map-of-oncomir-1-at-single-nucleotide-resolution
#19
Saikat Chakraborty, Yamuna Krishnan
The miR-17-92a cluster, also known as 'oncomiR-1', is an RNA transcript that plays a pivotal regulatory role in cellular processes, including the cell cycle, proliferation and apoptosis. Its dysregulation underlies the development of several cancers. Oncomir-1 comprises six constituent miRNAs, each processed with different efficiencies as a function of both developmental time and tissue type. The structural mechanisms that regulate such differential processing are unknown, and this has impeded our understanding of the dysregulation of oncomiR-1 in pathophysiology...
September 19, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28934475/in-vivo-probing-of-nascent-rna-structures-reveals-principles-of-cotranscriptional-folding
#20
Danny Incarnato, Edoardo Morandi, Francesca Anselmi, Lisa M Simon, Giulia Basile, Salvatore Oliviero
Defining the in vivo folding pathway of cellular RNAs is essential to understand how they reach their final native conformation. We here introduce a novel method, named Structural Probing of Elongating Transcripts (SPET-seq), that permits single-base resolution analysis of transcription intermediates' secondary structures on a transcriptome-wide scale, enabling base-resolution analysis of the RNA folding events. Our results suggest that cotranscriptional RNA folding in vivo is a mixture of cooperative folding events, in which local RNA secondary structure elements are formed as they get transcribed, and non-cooperative events, in which 5'-halves of long-range helices get sequestered into transient non-native interactions until their 3' counterparts have been transcribed...
September 19, 2017: Nucleic Acids Research
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