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https://www.readbyqxmd.com/read/28938432/mitochondrial-function-regulated-by-mitoguardin-1-2-is-crucial-for-ovarian-endocrine-functions-and-ovulation
#1
Xiao-Man Liu, Yin-Li Zhang, Shu-Yan Ji, Long-Wen Zhao, Wei-Na Shang, Dali Li, Zijiang Chen, Chao Tong, Heng-Yu Fan
The balances of mitochondrial dynamic changes, mitochondrial morphology, and mitochondrial number are critical in cell metabolism. Once disturbed, disorders in these processes generally cause diseases or even death in animals. We performed large-scale genetic screenings in fruit flies and discovered the new gene mitoguardin (miga) that encodes for a mitochondrial outer membrane protein. In order to examine the physiological functions of its mammalian homologs Miga1 and 2, we generated Miga1 and Miga2 single- and double-knockout mouse strains and found that the knockout mice were viable, but the females were subfertile...
August 23, 2017: Endocrinology
https://www.readbyqxmd.com/read/28938120/prion-like-domains-program-ewing-s-sarcoma
#2
James Shorter
Prion-like domains have emerged as important drivers of neurodegenerative disease. Now, Boulay et al. establish that the translocated prion-like domain of the oncogenic EWS-FLI1 fusion protein enables phase-separation events, which inappropriately recruit chromatin-remodeling factors to elicit the aberrant transcriptional programs underlying Ewing's sarcoma.
September 21, 2017: Cell
https://www.readbyqxmd.com/read/28937879/advax4-delta-inulin-combination-adjuvant-together-with-ecmx-a-fusion-construct-of-three-protective-mtb-antigens-induces-a-potent-th1-immune-response-and-protects-mice-against-mycobacterium-tuberculosis-infection
#3
Bruno de Paula Oliveira Santos, Monalisa Martins Trentini, Renato Beilner Machado, Mara Rúbia Nunes Celes, André Kipnis, Nikolai Petrovsky, Ana Paula Junqueira-Kipnis
Tuberculosis (TB) remains a main public health concern and 10.4 million new cases occurred in 2015 around the world. BCG is the only approved vaccine against TB, but has variable efficacy and new vaccines are needed. We developed two new mTB vaccine candidates based on the recombinant fusion proteins, rCMX and rECMXformulated with Advax4, a new combination adjuvant combining delta inulin, CpG oligonucleotide and murabutide. When BALB/c mice were immunized three times intramuscularly with these vaccine formulationsInjection of Advax4 alone increased the percentage of lymphatic endothelial cells and activated macrophages (F480/CD11b(+)) in the draining lymph nodes consistent with a chemotactic adjuvant effect...
September 22, 2017: Human Vaccines & Immunotherapeutics
https://www.readbyqxmd.com/read/28937754/bi-functional-elastin-like-polypeptide-nanoparticles-bind-rapamycin-and-integrins-and-suppress-tumor-growth-in-vivo
#4
Jugal P Dhandhukia, Pu Shi, Santosh Peddi, Zhe Li, Suhaas Rayudu Aluri, Yaping Ju, Dab Brill, Wan Wang, Siti Janib, Yi-An Lin, Shuanglong Liu, Honggang Cui, John Andrew MacKay
Recombinant protein-polymer scaffolds such as Elastin-Like Polypeptides (ELPs) offer drug delivery opportunities including biocompatibility, monodispersity, and multi-functionality. We recently reported that fusion of FK-506 binding protein 12 (FKBP) to an ELP nanoparticle (FSI) increases rapamycin (Rapa) solubility, suppresses growth of breast cancer xenografts, and reduces side-effects observed with free drug controls. This new report significantly advances this carrier strategy by demonstrating the co-assembly of two different ELP diblock copolymers containing drug-loading and tumor-targeting domains...
September 22, 2017: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/28936802/significance-of-mitochondrial-protein-post-translational-modifications-in-pathophysiology-of-brain-injury
#5
Nina Klimova, Aaron Long, Tibor Kristian
Mitochondria are complex organelles that undergo constant fusion and fission in order to adapt to the ever-changing cellular environment. The fusion/fission proteins, localized in the inner and outer mitochondrial membrane, play critical roles under pathological conditions such as acute brain injury and neurodegenerative diseases. Post-translational modifications of these proteins tightly regulate their function and activity, ultimately impacting mitochondrial dynamics and their efficiency to generate ATP. The individual post-translational modifications that are known to affect mitochondrial dynamics include SUMOylation, ubiquitination, phosphorylation, S-nitrosylation, acetylation, O-linked N-acetyl-glucosamine glycosylation, ADP-ribosylation, and proteolytic cleavage...
September 21, 2017: Translational Stroke Research
https://www.readbyqxmd.com/read/28936738/vascular-calcification-regulation-by-exosomes-in-the-vascular-wall
#6
Marcel Liberman, Luciana Cavalheiro Marti
Vascular calcification is a tightly regulated process that increases during ageing and occurs mainly in patients with diabetes and chronic renal failure. Exosomes are small membrane vesicles that are synthesized in a particular population of endosomes, also called multivesicular bodies, by inside budding into the lumen of the compartment. After fusion of exosome with the plasma membrane, these internal vesicles are secreted. Exosomes have a defined set of membrane and cytosolic proteins. The physiological function of exosomes is still a matter of debate...
2017: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/28936674/conditional-modulation-of-biological-processes-by-low-temperature-degrons
#7
Nico Dissmeyer
Conditional modulation of biological processes plays key roles in basic and applied research and in translation. It can be achieved on various levels via a multitude of approaches. One of the directions is manipulating target protein levels and activity by transcriptional, posttranscriptional, translational, and posttranslational control. Because in most of these techniques, the synthesis of the target proteins is adjusted to the needs, they all rely on the specific half-life of the target protein and its turn-over...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28936664/analysis-of-proteins-enriched-in-rice-gamete
#8
Takashi Okamoto
In angiosperms, female gamete differentiation, fertilization, and subsequent zygotic development occur in embryo sacs deeply embedded in the ovaries. Despite their importance in plant reproduction and development, how the egg cell is specialized, fuses with the sperm cell, and converts into an active zygote for early embryogenesis remains unclear. Proteins enriched in each cell type are thought to reflect the biological function of the cells, since the composition of cellular proteins generally differs depending on cell type...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28936650/analysis-of-fluorescent-reporter-activity-in-the-male-germline-during-pollen-development-by-confocal-microscopy
#9
David Twell, Lynette Brownfield
The male germline of flowering plants develops within the vegetative cell of the male gametophyte (pollen). The germline is established by asymmetric division of the microspore to form the generative cell. Mitotic division of the generative cell then produces the two sperm cells required for double fertilization. These differentiate to produce the proteins required for gamete attachment and fusion. An important aspect of understanding germline development is the characterization of germline gene expression...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28936467/peroxisome-motility-measurement-and-quantification-assay
#10
Jeremy Metz, Inês G Castro, Michael Schrader
Organelle movement, distribution and interaction contribute to the organisation of the eukaryotic cell. Peroxisomes are multifunctional organelles which contribute to cellular lipid metabolism and ROS homeostasis. They distribute uniformly in mammalian cells and move along microtubules via kinesin and dynein motors. Their metabolic cooperation with mitochondria and the endoplasmic reticulum (ER) is essential for the β-oxidation of fatty acids and the synthesis of myelin lipids and polyunsaturated fatty acids...
September 5, 2017: Bio-protocol
https://www.readbyqxmd.com/read/28935569/constitutive-cd40-signaling-in-dendritic-cells-limits-atherosclerosis-by-provoking-inflammatory-bowel-disease-and-ensuing-cholesterol-malabsorption
#11
Pascal Kusters, Tom Seijkens, Christina Bürger, Bart Legein, Holger Winkels, Marion Gijbels, Christian Barthels, Remy Bennett, Linda Beckers, Dorothee Atzler, Erik Biessen, Thomas Brocker, Christian Weber, Norbert Gerdes, Esther Lutgens
The co-stimulatory molecule CD40 is a major driver of atherosclerosis. It is expressed on a wide variety of cell types including mature dendritic cells (DCs) and is required for optimal T cell activation and expansion. It remains undetermined if and how CD40 on DCs impacts the pathogenesis of atherosclerosis. Here we examined the effects of constitutively active CD40 in DCs on atherosclerosis, using low-density lipoprotein-deficient (Ldlr(-/-)) bone-marrow chimeras that express a transgene containing an engineered latent membrane protein 1 (LMP)/CD40 fusion protein conferring constitutive CD40 signaling under control of the DC-specific CD11c promoter (DC-LMP1/CD40)...
September 18, 2017: American Journal of Pathology
https://www.readbyqxmd.com/read/28935566/construction-of-protein-functionalized-virus-like-particles-of-parvovirus-b19
#12
Ismael Bustos-Jaimes, Ricardo Arturo Soto-Román, Isabel Alejandra Gutiérrez-Landa, Josefina Valadez-García, Carla Lizbeth Segovia-Trinidad
Decoration of virus-like particles (VLPs) expands the repertory of functions these particles can display. In the last years, VLPs have successfully been used as scaffolds to present different molecules, frequently through the specific reaction of chemical groups on the surface of the particles, or by protein engineering when the presentation of peptides or proteins is the primary goal. VLPs of parvovirus B19 (B19V), have been previously produced in vitro and its stability and ability to assemble into hybrid particles composed of wild-type and chimeric proteins evidenced their potential as research tools...
September 18, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28934494/derivatives-of-bst-like-gss-polymerase-with-improved-processivity-and-inhibitor-tolerance
#13
Igor P Oscorbin, Ekaterina A Belousova, Ulyana A Boyarskikh, Aleksandr I Zakabunin, Evgeny A Khrapov, Maksim L Filipenko
At the moment, one of the actual trends in medical diagnostics is a development of methods for practical applications such as point-of-care testing, POCT or research tools, for example, whole genome amplification, WGA. All the techniques are based on using of specific DNA polymerases having strand displacement activity, high synthetic processivity, fidelity and, most significantly, tolerance to contaminants, appearing from analysed biological samples or collected under purification procedures. Here, we have designed a set of fusion enzymes based on catalytic domain of DNA polymerase I from Geobacillus sp...
September 19, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28934472/controlled-re-activation-of-epigenetically-silenced-tet-promoter-driven-transgene-expression-by-targeted-demethylation
#14
Natascha Gödecke, Lisha Zha, Shawal Spencer, Sara Behme, Pamela Riemer, Michael Rehli, Hansjörg Hauser, Dagmar Wirth
Faithful expression of transgenes in cell cultures and mice is often challenged by locus dependent epigenetic silencing. We investigated silencing of Tet-controlled expression cassettes within the mouse ROSA26 locus. We observed pronounced DNA methylation of the Tet promoter concomitant with loss of expression in mES cells as well as in differentiated cells and transgenic animals. Strikingly, the ROSA26 promoter remains active and methylation free indicating that this silencing mechanism specifically affects the transgene, but does not spread to the host's chromosomal neighborhood...
September 19, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28934205/a-rab5-gtpase-module-is-important-for-autophagosome
#15
Fan Zhou, Shenshen Zou, Yong Chen, Zhanna Lipatova, Dan Sun, Xiaolong Zhu, Rui Li, Zulin Wu, Weiming You, Xiaoxia Cong, Yiting Zhou, Zhiping Xie, Valeriya Gyurkovska, Yutao Liu, Qunli Li, Wenjing Li, Jie Cheng, Yongheng Liang, Nava Segev
In the conserved autophagy pathway, the double-membrane autophagosome (AP) engulfs cellular components to be delivered for degradation in the lysosome. While only sealed AP can productively fuse with the lysosome, the molecular mechanism of AP closure is currently unknown. Rab GTPases, which regulate all intracellular trafficking pathways in eukaryotes, also regulate autophagy. Rabs function in GTPase modules together with their activators and downstream effectors. In yeast, an autophagy-specific Ypt1 GTPase module, together with a set of autophagy-related proteins (Atgs) and a phosphatidylinositol-3-phosphate (PI3P) kinase, regulates AP formation...
September 21, 2017: PLoS Genetics
https://www.readbyqxmd.com/read/28933823/two-color-810-nm-sted-nanoscopy-of-living-cells-with-endogenous-snap-tagged-fusion-proteins
#16
Alexey N Butkevich, Haisen Ta, Michael Ratz, Stefan Stold, Stefan Jakobs, Vladimir N Belov, Stefan W Hell
A 810 nm STED nanoscopy setup and an appropriate combination of two fluorescent dyes (Si-rhodamine 680SiR and carbopyronine 610CP) have been developed for near-IR live-cell superresolution imaging. Vimentin endogenously tagged using the CRISPR/Cas9 approach with the SNAP-tag, together with a non-covalent tubulin label, provided reliable and cell-to-cell reproducible dual-color confocal and STED imaging of the cytoskeleton in living cells.
September 21, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/28933687/tagging-of-the-vaccinia-virus-protein-f13-with-mcherry-causes-aberrant-virion-morphogenesis
#17
David C J Carpentier, Michael S Hollinshead, Helen A Ewles, Stacey-Ann Lee, Geoffrey L Smith
Vaccinia virus produces two distinct infectious virions; the single-enveloped intracellular mature virus (IMV), which remains in the cell until cell lysis, and the double-enveloped extracellular enveloped virus (EEV), which mediates virus spread. The latter is derived from a triple-enveloped intracellular enveloped virus (IEV) precursor, which is transported to the cell periphery by the kinesin-1 motor complex. This transport involves the viral protein A36 as well as F12 and E2. A36 is an integral membrane protein associated with the outer virus envelope and is the only known direct link between virion and kinesin-1 complex...
September 20, 2017: Journal of General Virology
https://www.readbyqxmd.com/read/28933668/nasal-aluminum-oxy-hydroxide-enables-adsorbed-antigens-to-induce-specific-systemic-and-mucosal-immune-responses
#18
Haiyue Xu, Tinashe B Ruwona, Sachin G Thakkar, Yanping Chen, Mingtao Zeng, Zhengrong Cui
Some insoluble aluminum salts are commonly used in injectable vaccines as adjuvants to accelerate, prolong, or enhance the antigen-specific immune responses. Data from previous studies testing the nasal mucosal vaccine adjuvant activity of aluminum salts are conflicting. The present study is designed to further assess the feasibility of repurposing aluminum salts in injectable vaccines as nasal mucosal vaccine adjuvants. Using Alhydrogel®, the international scientific standard of aluminum (oxy)hydroxide gels, and ovalbumin or 3 × M2e-HA2, a synthetic influenza virus fusion protein, as antigens, we showed in a mouse model that when dosed intranasally Alhydrogel® enables antigens adsorbed on it to induce stronger antigen-specific immune responses in both serum samples (e...
September 21, 2017: Human Vaccines & Immunotherapeutics
https://www.readbyqxmd.com/read/28933638/autophagy-pathway-cellular-and-molecular-mechanisms
#19
Li Yu, Yang Chen, Sharon A Tooze
Macroautophagy/autophagy is an essential, conserved self-eating process that cells perform to allow degradation of intracellular components, including soluble proteins, aggregated proteins, organelles, macromolecular complexes, and foreign bodies. The process requires formation of a double-membrane structure containing the sequestered cytoplasmic material, the autophagosome, that ultimately fuses with the lysosome. This review will define this process and the cellular pathways required, from the formation of the double membrane to the fusion with lysosomes in molecular terms, and in particular highlight the recent progress in our understanding of this complex process...
September 21, 2017: Autophagy
https://www.readbyqxmd.com/read/28933316/photo-activatable-akt-probe-a-new-tool-to-study-the-akt-dependent-physiopathology-of-cancer-cells
#20
Sanae Haga, Takeaki Ozawa, Naoki Morita, Mami Asano, Shigeki Jin, Michitaka Ozakia
Akt is commonly overexpressed and activated in cancer cells, and plays a pivotal role in cell survival, protection andchemo-resistance. Therefore, Akt is one of the target molecules in understanding characters of cancer cells and developing anti-cancer drugs. Here, we examined whether a newly developed photo-activatable Akt (PA-Akt) probe, based on a light-inducible protein interaction module of plant cryptochrome2 (CRY2) and cryptochrome-interacting basic-helix-loop-helix (CIB1), can regulate Akt-associated cell functions...
August 30, 2017: Oncology Research
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