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https://www.readbyqxmd.com/read/30075102/overexpression-and-purification-of-human-myosins-from-transiently-and-stably-transfected-suspension-adapted-hek293sf-3f6-cells
#1
Marko Ušaj, Lilach Zattelman, Ronit Regev, Boris I Shneyer, Naama Wiesel-Motiuk, Arnon Henn
The myosin family of motor proteins is an attractive target of therapeutic small-molecule protein inhibitors and modulators. Milligrams of protein quantities are required to conduct proper biophysical and biochemical studies to understand myosin functions. Myosin protein expression and purification represent a critical starting point towards this goal. Established utilization of Dictyostelium discoideum, Drosophila melanogaster, insect and mouse cells for myosin expression and purification is limited, cost, labor and time inefficient particularly for (full-length) human myosins...
July 31, 2018: Analytical Biochemistry
https://www.readbyqxmd.com/read/30072914/cloning-and-characterization-of-two-toll-receptors-pctoll5-and-pctoll6-in-response-to-white-spot-syndrome-virus-in-the-red-swamp-crayfish-procambarus-clarkii
#2
Ying Huang, Yihong Chen, Kaimin Hui, Qian Ren
Toll/Toll-like receptors are key components in the innate immune responses of invertebrates. In this study, we identified two novel Toll receptors ( PcToll5 and PcToll6 ) from the red swamp crayfish Procambarus clarkii . The complete cDNA sequence of PcToll5 is 4247 bp, encoding a 1293 amino acid polypeptide. The full-length 4688 bp PcToll6 encodes a putative protein of 1195 amino acids. Quantitative RT-PCR analysis indicated that PcToll5 and PcToll6 were constitutively expressed in all tissues studied. The highest expression levels of PcToll5 and PcToll6 were found in the intestine and gills, respectively, and were significantly upregulated from 24 to 48 h during white spot syndrome virus (WSSV) challenge...
2018: Frontiers in Physiology
https://www.readbyqxmd.com/read/30061855/modeling-of-myotonic-dystrophy-cardiac-phenotypes-in-drosophila
#3
REVIEW
Mouli Chakraborty, Beatriz Llamusi, Ruben Artero
After respiratory distress, cardiac dysfunction is the second most common cause of fatality associated with the myotonic dystrophy (DM) disease. Despite the prevalance of heart failure in DM, physiopathological studies on heart symptoms have been relatively scarce because few murine models faithfully reproduce the cardiac disease. Consequently, only a small number of candidate compounds have been evaluated in this specific phenotype. To help cover this gap Drosophila combines the amenability of its invertebrate genetics with the possibility of quickly acquiring physiological parameters suitable for meaningful comparisons with vertebrate animal models and humans...
2018: Frontiers in Neurology
https://www.readbyqxmd.com/read/30059535/intersection-of-phosphate-transport-oxidative-stress-and-tor-signalling-in-candida-albicans-virulence
#4
Ning-Ning Liu, Priya Uppuluri, Achille Broggi, Angelique Besold, Kicki Ryman, Hiroto Kambara, Norma Solis, Viola Lorenz, Wanjun Qi, Maikel Acosta-Zaldívar, S Noushin Emami, Bin Bao, Dingding An, Francisco A Bonilla, Martha Sola-Visner, Scott G Filler, Hongbo R Luo, Ylva Engström, Per Olof Ljungdahl, Valeria C Culotta, Ivan Zanoni, Jose L Lopez-Ribot, Julia R Köhler
Phosphate is an essential macronutrient required for cell growth and division. Pho84 is the major high-affinity cell-surface phosphate importer of Saccharomyces cerevisiae and a crucial element in the phosphate homeostatic system of this model yeast. We found that loss of Candida albicans Pho84 attenuated virulence in Drosophila and murine oropharyngeal and disseminated models of invasive infection, and conferred hypersensitivity to neutrophil killing. Susceptibility of cells lacking Pho84 to neutrophil attack depended on reactive oxygen species (ROS): pho84-/- cells were no more susceptible than wild type C...
July 2018: PLoS Pathogens
https://www.readbyqxmd.com/read/30057203/nona-and-cpx-link-the-circadian-clockwork-to-locomotor-activity-in-drosophila
#5
Weifei Luo, Fang Guo, Aoife McMahon, Shalise Couvertier, Hua Jin, Madelen Diaz, Allegra Fieldsend, Eranthie Weerapana, Michael Rosbash
Drosophila NonA and its mammalian ortholog NONO are members of the Drosophila behavior and human splicing (DBHS) family. NONO also has a strong circadian connection: it associates with the circadian repressor protein PERIOD (PER) and contributes to circadian timekeeping. Here, we investigate NonA, which is required for proper levels of evening locomotor activity as well as a normal free-running period in Drosophila. NonA is associated with the positive transcription factor CLOCK/CYCLE (CLK/CYC), interacts directly with complexin (cpx) pre-mRNA, and upregulates gene expression, including the gene cpx...
July 24, 2018: Neuron
https://www.readbyqxmd.com/read/30050143/npm-hmlf1-fusion-protein-suppresses-defects-of-a-drosophila-ftld-model-expressing-the-human-fus-gene
#6
Itaru Yamamoto, Yumiko Azuma, Yukie Kushimura, Hideki Yoshida, Ikuko Mizuta, Toshiki Mizuno, Morio Ueyama, Yoshitaka Nagai, Takahiko Tokuda, Masamitsu Yamaguchi
Fused in sarcoma (FUS) was identified as a component of typical inclusions in frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS). In FTLD, both nuclear and cytoplasmic inclusions with wild-type FUS exist, while cytoplasmic inclusions with a mutant-form of FUS occur in many ALS cases. These observations imply that FUS plays a role across these two diseases. In this study, we examined the effect of several proteins including molecular chaperons on the aberrant eye morphology phenotype induced by overexpression of wild-type human FUS (hFUS) in Drosophila eye imaginal discs...
July 26, 2018: Scientific Reports
https://www.readbyqxmd.com/read/30032358/drosophila-hsp67bc-hot-spot-variants-alter-muscle-structure-and-function
#7
Jadwiga Jabłońska, Magda Dubińska-Magiera, Teresa Jagla, Krzysztof Jagla, Małgorzata Daczewska
The Drosophila Hsp67Bc gene encodes a protein belonging to the small heat-shock protein (sHSP) family, identified as the nearest functional ortholog of human HSPB8. The most prominent activity of sHSPs is preventing the irreversible aggregation of various non-native polypeptides. Moreover, they are involved in processes such as development, aging, maintenance of the cytoskeletal architecture and autophagy. In larval muscles Hsp67Bc localizes to the Z- and A-bands, which suggests its role as part of the conserved chaperone complex required for Z-disk maintenance...
July 21, 2018: Cellular and Molecular Life Sciences: CMLS
https://www.readbyqxmd.com/read/30031756/ctbp-represses-dpp-dependent-mad-activation-during-drosophila-eye-development
#8
Nadia Eusebio, Lígia Tavares, Paulo S Pereira
Complex networks of signaling pathways maintain the correct balance between positive and negative growth signals, ensuring that tissues achieve proper sizes and differentiation pattern during development. In Drosophila, Dpp, a member of the TGFβ family, plays two main roles during larval eye development. In the early eye primordium, Dpp promotes growth and cell survival, but later on, it switches its function to induce a developmentally-regulated cell cycle arrest in the G1 phase and neuronal photoreceptor differentiation...
July 19, 2018: Developmental Biology
https://www.readbyqxmd.com/read/30030398/the-drosophila-receptor-tyrosine-kinase-alk-constrains-long-term-memory-formation
#9
Jean Y Gouzi, Mikela Bouraimi, Ilianna G Roussou, Anastasios Moressis, Efthimios M C Skoulakis
In addition to mechanisms promoting protein-synthesis dependent long-term memory (PSD-LTM), the process appears to also be specifically constrained. We present evidence that the highly conserved Receptor Tyrosine Kinase dAlk is a novel PSD-LTM attenuator in Drosophila. Reduction of dAlk levels in adult α/β mushroom body (MB) neurons during conditioning elevates LTM, whereas its overexpression impairs it. Unlike other memory suppressor proteins and miRNAs, dAlk within the MBs constrains PSD-LTM specifically, but constrains learning outside the MBs as previously shown...
July 20, 2018: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
https://www.readbyqxmd.com/read/30026307/planar-cell-polarity-gene-fuz-triggers-apoptosis-in-neurodegenerative-disease-models
#10
Zhefan Stephen Chen, Li Li, Shaohong Peng, Francis M Chen, Qian Zhang, Ying An, Xiao Lin, Wen Li, Alex Chun Koon, Ting-Fung Chan, Kwok-Fai Lau, Jacky Chi Ki Ngo, Wing Tak Wong, Kin Ming Kwan, Ho Yin Edwin Chan
Planar cell polarity (PCP) describes a cell-cell communication process through which individual cells coordinate and align within the plane of a tissue. In this study, we show that overexpression of Fuz , a PCP gene, triggers neuronal apoptosis via the dishevelled/Rac1 GTPase/MEKK1/JNK/caspase signalling axis. Consistent with this finding, endogenous Fuz expression is upregulated in models of polyglutamine (polyQ) diseases and in fibroblasts from spinocerebellar ataxia type 3 (SCA3) patients. The disruption of this upregulation mitigates polyQ-induced neurodegeneration in Drosophila We show that the transcriptional regulator Yin Yang 1 (YY1) associates with the Fuz promoter...
July 19, 2018: EMBO Reports
https://www.readbyqxmd.com/read/30011269/modulation-of-mir-210-alters-phasing-of-circadian-locomotor-activity-and-impairs-projections-of-pdf-clock-neurons-in-drosophila-melanogaster
#11
Paola Cusumano, Alberto Biscontin, Federica Sandrelli, Gabriella M Mazzotta, Claudia Tregnago, Cristiano De Pittà, Rodolfo Costa
Single microRNAs are usually associated with hundreds of putative target genes that can influence multiple phenotypic traits in Drosophila, ranging from development to behaviour. We investigated the function of Drosophila miR-210 in circadian behaviour by misexpressing it within circadian clock cells. Manipulation of miR-210 expression levels in the PDF (pigment dispersing factor) positive neurons affected the phase of locomotor activity, under both light-dark conditions and constant darkness. PER cyclical expression was not affected in clock neurons, however, when miR-210 was up-regulated, a dramatic alteration in the morphology of PDF ventral lateral neuron (LNv) arborisations was observed...
July 2018: PLoS Genetics
https://www.readbyqxmd.com/read/30002614/asiatic-acid-prevents-oxidative-stress-and-apoptosis-by-inhibiting-the-translocation-of-%C3%AE-synuclein-into-mitochondria
#12
Hongqun Ding, Yuyun Xiong, Jing Sun, Chen Chen, Jing Gao, Huaxi Xu
The association of α-synuclein (α-syn) with mitochondria occurs through interaction with mitochondrial complex I. Defects in this protein have been linked to the pathogenesis of Parkinson disease (PD). Overexpression of α-synuclein in cells has been suggested to cause elevations in mitochondrial oxidant radicals and structural and functional abnormalities in mitochondria. Asiatic acid (AA), a triterpenoid, is an antioxidant that is used for depression, and we have shown that pretreatment with AA can prevent PD-like damage, but its therapeutic effects in PD and mechanism remain unknown...
2018: Frontiers in Neuroscience
https://www.readbyqxmd.com/read/30001323/a-bidirectional-relationship-between-sleep-and-oxidative-stress-in-drosophila
#13
Vanessa M Hill, Reed M O'Connor, Gunter B Sissoko, Ifeoma S Irobunda, Stephen Leong, Julie C Canman, Nicholas Stavropoulos, Mimi Shirasu-Hiza
Although sleep appears to be broadly conserved in animals, the physiological functions of sleep remain unclear. In this study, we sought to identify a physiological defect common to a diverse group of short-sleeping Drosophila mutants, which might provide insight into the function and regulation of sleep. We found that these short-sleeping mutants share a common phenotype of sensitivity to acute oxidative stress, exhibiting shorter survival times than controls. We further showed that increasing sleep in wild-type flies using genetic or pharmacological approaches increases survival after oxidative challenge...
July 2018: PLoS Biology
https://www.readbyqxmd.com/read/29995586/atp6ap2-functions-as-a-v-atpase-assembly-factor-in-the-endoplasmic-reticulum
#14
Maria Clara Guida, Tobias Hermle, Laurie A Graham, Virginie Hauser, Margret Ryan, Tom H Stevens, Matias Simons
ATP6AP2 (also known as the (pro)renin receptor) is a type I transmembrane protein that can be cleaved into two fragments in the Golgi apparatus. While in Drosophila ATP6AP2 functions in the planar cell polarity (PCP) pathway, recent human genetic studies have suggested that ATP6AP2 could participate in the assembly of the V-ATPase in the endoplasmic reticulum (ER). Using a yeast model, we show here that the V-ATPase assembly factor Voa1 can functionally be replaced by Drosophila ATP6AP2. This rescue is even more efficient when co-expressing its binding partner ATP6AP1, indicating that these two proteins together fulfill Voa1 functions in higher organisms...
July 11, 2018: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/29989152/transplantation-of-photoreceptor-precursor-cells-into-the-retina-of-an-adult-drosophila
#15
Takahisa Suzuki, Keita Oochi, Satoko Hakeda-Suzuki, Takashi Suzuki
Blindness caused by the disconnection between photoreceptor cells and the brain can be cured by restoring this connection through the transplantation of retinal precursor neurons. However, even after transplanting these cells, it is still unclear how to guide the axons over the long distance from the retina to the brain. To establish a method of guiding the axons of transplanted neurons, we used the Drosophila visual system. By testing different conditions, including the dissociation and preincubation length, we have successfully established a method to transplant photoreceptor precursor cells isolated from the developing eye discs of third-instar larvae into the adult retina...
July 10, 2018: Development, Growth & Differentiation
https://www.readbyqxmd.com/read/29973396/cultured-cells-and-wing-disc-size-of-silkworm-can-be-controlled-by-the-hippo-pathway
#16
Zi Liang, Yahong Lu, Ying Qian, Liyuan Zhu, Sulan Kuang, Fei Chen, Yongjie Feng, Xiaolong Hu, Guangli Cao, Renyu Xue, Chengliang Gong
Hippo signalling represents a cell proliferation and organ-size control pathway. Yorki (Yki), a component of the Hippo pathway, induces the transcription of a number of targets that promote cell proliferation and survival. The functions of Yki have been characterized in Drosophila and mammals, while there are few reports on silkworm, Bombyx mori In the present study, we found that BmYki3 facilitates cell migration and cell division, and enlarges the cultured cell and wing disc size. Co-immunoprecipitation results indicated that BmYki3 may interact with thymosin, E3 ubiquitin-protein ligase, protein kinase ASK1, dedicator of cytokinesis protein 1, calcium-independent phospholipase A2 and beta-spectrin...
July 2018: Open Biology
https://www.readbyqxmd.com/read/29946036/-drosophila-model-of-myosin-myopathy-rescued-by-overexpression-of-a-trim-protein-family-member
#17
Martin Dahl-Halvarsson, Montse Olive, Malgorzata Pokrzywa, Katarina Ejeskär, Ruth H Palmer, Anne Elisabeth Uv, Homa Tajsharghi
Myosin is a molecular motor indispensable for body movement and heart contractility. Apart from pure cardiomyopathy, mutations in MYH7 encoding slow/β-cardiac myosin heavy chain also cause skeletal muscle disease with or without cardiac involvement. Mutations within the α-helical rod domain of MYH7 are mainly associated with Laing distal myopathy. To investigate the mechanisms underlying the pathology of the recurrent causative MYH7 mutation (K1729del), we have developed a Drosophila melanogaster model of Laing distal myopathy by genomic engineering of the Drosophila Mhc locus...
July 10, 2018: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29938216/dstim-and-ral-exocyst-mediated-synaptic-release-from-pupal-dopaminergic-neurons-sustains-drosophila-flight
#18
Shlesha Richhariya, Siddharth Jayakumar, Sanjay Kumar Sukumar, Gaiti Hasan
Manifestation of appropriate behavior in adult animals requires developmental mechanisms that help in the formation of correctly wired neural circuits. Flight circuit development in Drosophila requires store-operated calcium entry (SOCE) through the STIM/Orai pathway. SOCE-associated flight deficits in adult Drosophila derive extensively from regulation of gene expression in pupal neurons, and one such SOCE-regulated gene encodes the small GTPase Ral . The cellular mechanism by which Ral helps in maturation of the flight circuit was not understood...
May 2018: ENeuro
https://www.readbyqxmd.com/read/29935285/sptoll1-and-sptoll2-modulate-the-expression-of-antimicrobial-peptides-in-scylla-paramamosain
#19
Yan Chen, Jude Juventus Aweya, Wanwei Sun, Xiaoyuan Wei, Yi Gong, Hongyu Ma, Yueling Zhang, Xiaobo Wen, Shengkang Li
Tolls and Toll-like receptors (TLRs) were the first pattern recognition receptors (PRRs) identified to play key roles in host innate immunity. However, relatively little is known about other types of Toll-like receptors in Scylla paramamosain, although a Toll-like receptor (SpToll1) has recently been cloned. In this study, we cloned and characterized another novel Toll-like receptor 2 (SpToll2) from S. paramamosain. The full-length cDNA of SpToll2 is 3391 bp with a 2646 bp open reading frame (ORF) encoding a putative protein of 881 amino acids, and predicted to contain six extracellular leucine-rich repeat (LRR) domains, a transmembrane domain and an intracellular Toll/IL-1 receptor (TIR) domain...
October 2018: Developmental and Comparative Immunology
https://www.readbyqxmd.com/read/29931724/overexpression-of-a-glutathione-s-transferase-mdgst-and-a-galactosyltransferase-like-gene-mdgt1-is-responsible-for-imidacloprid-resistance-in-house-flies
#20
William R Reid, Haina Sun, James J Becnel, Andrew G Clark, Jeffrey G Scott
BACKGROUND: Neonicotinoids are the largest class of insecticides and are used for control of house fly populations at animal production facilities throughout the world. There have been several reports of neonicotinoid resistance in house fly populations, but identification of the factors involved in resistance has proven challenging. The KS8S3 population of house flies is highly resistant to the neonicotinoid insecticide imidacloprid due to two factors: one on chromosome 3 and one on chromosome 4...
June 21, 2018: Pest Management Science
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