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https://www.readbyqxmd.com/read/28431041/bidirectional-approaches-for-optogenetic-regulation-of-gene-expression-in-mammalian-cells-using-arabidopsis-cryptochrome-2
#1
Gopal P Pathak, Jessica I Spiltoir, Camilla Höglund, Lauren R Polstein, Sari Heine-Koskinen, Charles A Gersbach, Jari Rossi, Chandra L Tucker
Optogenetic tools allow regulation of cellular processes with light, which can be delivered with spatiotemporal resolution. In previous work, we used cryptochrome 2 (CRY2) and CIB1, Arabidopsis proteins that interact upon light illumination, to regulate transcription with light in yeast. While adopting this approach to regulate transcription in mammalian cells, we observed light-dependent redistribution and clearing of CRY2-tethered proteins within the nucleus. The nuclear clearing phenotype was dependent on the presence of a dimerization domain contained within the CRY2-fused transcriptional activators...
April 20, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28412365/vaccinia-related-kinase-3-vrk3-sets-the-circadian-period-and-amplitude-by-affecting-the-subcellular-localization-of-clock-proteins-in-mammalian-cells
#2
Nayoung Park, Ji-Eun Song, Seongsu Jeong, Thanh Thi Tran, Hyuk Wan Ko, Eun Young Kim
In the eukaryotic circadian clock machinery, negative feedback repression of CLOCK (CLK) and BMAL1 transcriptional activity by PERIOD (PER) and CRYPTOCHROME (CRY) underlies the basis for 24 h rhythmic gene expression. Thus, precise regulation of the time-dependent nuclear entry of circadian repressors is crucial to generating normal circadian rhythms. Here, we sought to identify novel kinase(s) that regulate nuclear entry of mammalian CRY1 (mCRY1) with an unbiased screening using red fluorescent protein (RFP)-tagged human kinome expression plasmids in mammalian cells...
April 12, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28409381/dash-type-cryptochromes-regulate-fruiting-body-development-and-secondary-metabolism-differently-than-cmwc-1-in-the-fungus-cordyceps-militaris
#3
Fen Wang, Xinhua Song, Xiaoming Dong, Jiaojiao Zhang, Caihong Dong
Cryptochromes (CRYs) belong to the photolyase/cryptochrome flavoprotein family, which is widely distributed in all kingdoms. A phylogenetic analysis indicated that three Cordyceps militaris proteins [i.e., cryptochrome DASH (CmCRY-DASH), (6-4) photolyase, and cyclobutane pyrimidine dimer (CPD) class I photolyase] belong to separate fungal photolyase/cryptochrome subfamilies. CmCRY-DASH consists of DNA photolyase and flavin adenine dinucleotide-binding domains, with RGG repeats in a C-terminal extension. Considerably, more carotenoids and cordycepin accumulated in the ΔCmcry-DASH strain than in the wild-type or ΔCmwc-1 strains, indicating an inhibitory role for CmCRY-DASH in these biosynthetic pathways...
April 13, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28405468/cryptochrome-genes-form-an-oscillatory-loop-independent-of-the-per-tim-loop-in-the-circadian-clockwork-of-the-cricket-gryllus-bimaculatus
#4
Atsushi Tokuoka, Taichi Q Itoh, Shinryo Hori, Outa Uryu, Yoshiki Danbara, Motoki Nose, Tetsuya Bando, Teiichi Tanimura, Kenji Tomioka
BACKGROUND: Animals exhibit circadian rhythms with a period of approximately 24 h in various physiological functions, including locomotor activity. This rhythm is controlled by an endogenous oscillatory mechanism, or circadian clock, which consists of cyclically expressed clock genes and their product proteins. cryptochrome (cry) genes are thought to be involved in the clock mechanism, and their functions have been examined extensively in holometabolous insects, but in hemimetabolous insects their role is less well understood...
2017: Zoological Letters
https://www.readbyqxmd.com/read/28402850/fad-regulates-cryptochrome-protein-stability-and-circadian-clock-in-mice
#5
Arisa Hirano, Daniel Braas, Ying-Hui Fu, Louis J Ptáček
The circadian clock generates biological rhythms of metabolic and physiological processes, including the sleep-wake cycle. We previously identified a missense mutation in the flavin adenine dinucleotide (FAD) binding pocket of CRYPTOCHROME2 (CRY2), a clock protein that causes human advanced sleep phase. This prompted us to examine the role of FAD as a mediator of the clock and metabolism. FAD stabilized CRY proteins, leading to increased protein levels. In contrast, knockdown of Riboflavin kinase (Rfk), an FAD biosynthetic enzyme, enhanced CRY degradation...
April 11, 2017: Cell Reports
https://www.readbyqxmd.com/read/28396831/cryptochrome-2-extensively-regulates-transcription-of-the-chloroplast-genome-in-tomato
#6
Paolo Facella, Fabrizio Carbone, Antonio Placido, Gaetano Perrotta
Light plays a key role in the regulation of many physiological processes required for plant and chloroplast development. Plant cryptochromes (crys) play an important role in monitoring, capturing, and transmitting the light stimuli. In this study, we analyzed the effects of CRY2 overexpression on transcription of tomato chloroplast genome by a tiling array, containing about 90 000 overlapping probes (5-nucleotide resolution). We profiled transcription in leaves of wild-type and CRY2-overexpressing plants grown in a diurnal cycle, to generate a comprehensive map of chloroplast transcription and to monitor potential specific modulations of the chloroplast transcriptome induced by the overexpression of CRY2...
April 2017: FEBS Open Bio
https://www.readbyqxmd.com/read/28393477/residues-at-a-single-site-differentiate-animal-cryptochromes-from-cyclobutane-pyrimidine-dimer-photolyases-by-affecting-the-proteins-preferences-for-reduced-fad
#7
Lei Xu, Bin Wen, Yuan Wang, Changqing Tian, Mingcai Wu, Guoping Zhu
Cryptochromes (CRYs) and photolyases belong to the cryptochrome/photolyase family (CPF). Reduced FAD is essential for photolyases to photorepair UV induced cyclobutane pyrimidine dimers (CPDs) or 6-4 photoproducts in DNA. In Drosophila CRY (dCRY, a type I animal CRY), FAD is converted to the anionic radical but not to the reduced state during illumination, which may induce a conformational change in the protein to relay the light signal downstream. To explore the foundation of these differences, multiple sequence alignment of 650 CPF protein sequences was performed...
April 10, 2017: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/28360843/magr-alone-is-insufficient-to-confer-cellular-calcium-responses-to-magnetic-stimulation
#8
Keliang Pang, He You, Yanbo Chen, Pengcheng Chu, Meiqin Hu, Jianying Shen, Wei Guo, Can Xie, Bai Lu
Magnetic manipulation of cell activity offers advantages over optical manipulation but an ideal tool remains elusive. The MagR protein was found through its interaction with cryptochrome (Cry) and the protein in solution appeared to respond to magnetic stimulation (MS). After we initiated an investigation on the specific role of MagR in cellular response to MS, a subsequent study claimed that MagR expression alone could achieve cellular activation by MS. Here we report that despite systematically testing different ways of measuring intracellular calcium and different MS protocols, it was not possible to detect any cellular or neuronal responses to MS in MagR-expressing HEK cells or primary neurons from the dorsal root ganglion and the hippocampus...
2017: Frontiers in Neural Circuits
https://www.readbyqxmd.com/read/28360233/a-plant-cryptochrome-controls-key-features-of-the-chlamydomonas-circadian-clock-and-its-life-cycle
#9
Nico Müller, Sandra Wenzel, Yong Zou, Sandra Künzel, Severin Sasso, Daniel Weiß, Katja Prager, Arthur R Grossman, Tilman Kottke, Maria Mittag
Cryptochromes are flavin binding proteins that act as blue light receptors in bacteria, fungi, plants and insects and are components of the circadian oscillator in mammals. Animal and plant cryptochromes are evolutionarily divergent, although the unicellular alga Chlamydomonas reinhardtii (Chlamydomonas throughout) has both an animal-like cryptochrome and a plant cryptochrome (pCRY, formerly designated CPH1). Here, we show that pCRY protein accumulates at night as part of a complex. Functional characterization of pCRY was performed based on an insertional mutant that expresses only 11% of the wild-type pCRY level...
March 30, 2017: Plant Physiology
https://www.readbyqxmd.com/read/28356366/zebra-finches-have-a-light-dependent-magnetic-compass-similar-to-migratory-birds
#10
Atticus Pinzon-Rodriguez, Rachel Muheim
Birds have a light-dependent magnetic compass that provides information about the spatial alignment of the geomagnetic field. It is proposed to be located in the avian retina and mediated by a light-induced, radical-pair mechanism involving cryptochromes as sensory receptor molecules. To investigate how the behavioural responses of birds under different light spectra match with cryptochromes as the primary magnetoreceptor, we examined the spectral properties of the magnetic compass in zebra finches. We trained birds to relocate a food reward in a spatial orientation task using magnetic compass cues...
April 1, 2017: Journal of Experimental Biology
https://www.readbyqxmd.com/read/28335761/the-effects-of-jiao-tai-wan-on-sleep-inflammation-and-insulin-resistance-in-obesity-resistant-rats-with-chronic-partial-sleep-deprivation
#11
Xin Zou, Wenya Huang, Fuer Lu, Ke Fang, Dingkun Wang, Shuyong Zhao, Jiming Jia, Lijun Xu, Kaifu Wang, Nan Wang, Hui Dong
BACKGROUND: Jiao-Tai-Wan (JTW), composed of Rhizome Coptidis and Cortex Cinnamomi, is a classical traditional Chinese prescription for treating insomnia. Several in vivo studies have concluded that JTW could exert its therapeutical effect in insomnia rats. However, the specific mechanism is still unclear. The present study aimed to explore the effect of JTW on sleep in obesity-resistant (OR) rats with chronic partial sleep deprivation (PSD) and to clarify its possible mechanism. METHODS: JTW was prepared and the main components contained in the granules were identified by 3D-High Performance Liquid Chromatography (3D-HPLC) assay...
March 23, 2017: BMC Complementary and Alternative Medicine
https://www.readbyqxmd.com/read/28317918/vertebrate-cryptochromes-are-vestigial-flavoproteins
#12
Roger J Kutta, Nataliya Archipowa, Linus O Johannissen, Alex R Jones, Nigel S Scrutton
All cryptochromes are currently classified as flavoproteins. In animals their best-described role is as components of the circadian clock. This circadian function is variable, and can be either light-dependent or -independent; the molecular origin of this difference is unknown. Type I animal cryptochromes are photoreceptors that entrain an organism's clock to its environment, whereas Type II (including mammals) regulate circadian timing in a light-independent manner. Here, we reveal that, in contrast to Type I, Type II animal cryptochromes lack the structural features to securely bind the photoactive flavin cofactor...
March 20, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28296939/can-a-hybrid-chemical-ferromagnetic-model-of-the-avian-compass-explain-its-outstanding-sensitivity-to-magnetic-noise
#13
Kirill Kavokin
While many properties of the magnetic compass of migratory birds are satisfactory explained within the chemical model of magnetoreception, its extreme sensitivity to radio-frequency magnetic fields remains a mystery. Apparently, this difficulty could be overcome if the magnetoreceptor model were augmented with a magnetite nanoparticle, which would amplify the magnetic field at the position of the magneto-sensitive cryptochrome molecule. However, comparison of the radio-frequency power used in the experiment with intrinsic magnetization noise of such a particle, estimated from the theory of fluctuations, shows that the required sensitivity cannot be reached with realistic parameters of iron-oxide nanocrystals...
2017: PloS One
https://www.readbyqxmd.com/read/28296892/blue-light-induced-accumulation-of-reactive-oxygen-species-is-a-consequence-of-the-drosophila-cryptochrome-photocycle
#14
Louis-David Arthaut, Nathalie Jourdan, Ali Mteyrek, Maria Procopio, Mohamed El-Esawi, Alain d'Harlingue, Pierre-Etienne Bouchet, Jacques Witczak, Thorsten Ritz, André Klarsfeld, Serge Birman, Robert J Usselman, Ute Hoecker, Carlos F Martino, Margaret Ahmad
Cryptochromes are evolutionarily conserved blue-light absorbing flavoproteins which participate in many important cellular processes including in entrainment of the circadian clock in plants, Drosophila and humans. Drosophila melanogaster cryptochrome (DmCry) absorbs light through a flavin (FAD) cofactor that undergoes photoreduction to the anionic radical (FAD•-) redox state both in vitro and in vivo. However, recent efforts to link this photoconversion to the initiation of a biological response have remained controversial...
2017: PloS One
https://www.readbyqxmd.com/read/28293892/light-regulated-protein-kinases-based-on-the-cry2-cib1-system
#15
Wignand W D Mühlhäuser, Maximilian Hörner, Wilfried Weber, Gerald Radziwill
Optogenetic approaches enable the control of biological processes in a time- and space-resolved manner. These light-based methods are noninvasive and by using light as sole activator minimize side effects in contrast to chemical inducers. Here, we provide a protocol for the targeted control of the activity of protein kinases in mammalian cells based on the photoreceptor cryptochrome 2 (CRY2) of Arabidopsis thaliana and its interaction partner CIB1. Blue light (450 nm)-induced binding of CRY2 to CIB1 allows the recruitment of a chimeric cytosolic protein kinase AKT1 to the plasma membrane accompanied with stimulation of its kinase activity...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28273503/resistance-of-arabidopsis-thaliana-l-photosynthetic-apparatus-to-uv-b-is-reduced-by-deficit-of-phytochromes-b-and-a
#16
Aleksandra Yu Khudyakova, Vladimir D Kreslavski, Galina N Shirshikova, Sergey K Zharmukhamedov, Anatoly A Kosobryukhov, Suleyman I Allakhverdiev
The photosynthetic responses of 25-day-old Arabidopsis phyA phyB double mutant (DM) compared with the wild type (WT) to UV-B radiation (1Wm(-2), 30min) were investigated. UV-B irradiation led to reduction of photosystem 2 (PS-2) activity and the photosynthetic rate. In plants grown under both white and red light (λm - 660nm) the reduction was greater in DM plants compared to the WT. Without UV-B irradiation a decrease in PS-2 activity was observed in DM grown under RL only. It is assumed that the lower content of UV-absorbing pigments and carotenoids observed in DM may be one of the reasons of reduced PS-2 resistance to UV-B...
April 2017: Journal of Photochemistry and Photobiology. B, Biology
https://www.readbyqxmd.com/read/28272509/optogenetic-control-of-the-dab1-signaling-pathway
#17
Liang Wang, Jonathan A Cooper
The Reelin-Dab1 signaling pathway regulates development of the mammalian brain, including neuron migrations in various brain regions, as well as learning and memory in adults. Extracellular Reelin binds to cell surface receptors and activates phosphorylation of the intracellular Dab1 protein. Dab1 is required for most effects of Reelin, but Dab1-independent pathways may contribute. Here we developed a single-component, photoactivatable Dab1 (opto-Dab1) by using the blue light-sensitive dimerization/oligomerization property of A...
March 8, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28271102/switching-from-adduct-formation-to-electron-transfer-in-a-light-oxygen-voltage-domain-containing-the-reactive-cysteine
#18
Kathrin Magerl, Ivan Stambolic, Bernhard Dick
LOV (light-, oxygen- or voltage-sensitive) domains act as photosensory units of many prokaryotic and eukaryotic proteins. Upon blue light excitation they undergo a photocycle via the excited triplet state of their flavin chromophore yielding the flavin-cysteinyl adduct. Adduct formation is highly conserved among all LOV domains and constitutes the primary step of LOV domain signaling. But recently, it has been shown that signal propagation can also be triggered by flavin photoreduction to the neutral semiquinone offering new prospects for protein engineering...
March 8, 2017: Physical Chemistry Chemical Physics: PCCP
https://www.readbyqxmd.com/read/28262491/adaptation-of-circadian-neuronal-network-to-photoperiod-in-high-latitude-european-drosophilids
#19
Pamela Menegazzi, Elena Dalla Benetta, Marta Beauchamp, Matthias Schlichting, Ingolf Steffan-Dewenter, Charlotte Helfrich-Förster
The genus Drosophila contains over 2,000 species that, stemming from a common ancestor in the Old World Tropics, populate today very different environments [1, 2] (reviewed in [3]). We found significant differences in the activity pattern of Drosophila species belonging to the holarctic virilis group, i.e., D. ezoana and D. littoralis, collected in Northern Europe, compared to that of the cosmopolitan D. melanogaster, collected close to the equator. These behavioral differences might have been of adaptive significance for colonizing high-latitude habitats and hence adjust to long photoperiods...
March 20, 2017: Current Biology: CB
https://www.readbyqxmd.com/read/28224616/critical-cholangiocarcinogenesis-control-by-cryptochrome-clock-genes
#20
Ali Mteyrek, Elisabeth Filipski, Catherine Guettier, Malgorzata Oklejewicz, Gijsbertus T J van der Horst, Alper Okyar, Francis Lévi
A coordinated network of molecular circadian clocks in individual cells generates 24-hr rhythms in liver metabolism and proliferation. Circadian disruption through chronic jet lag or Per2 clock gene mutation was shown to accelerate hepatocarcinoma development in mice. As divergent effects were reported for clock genes Per and Cry regarding xenobiotic toxicity, we questioned the role of Cry1 and Cry2 in liver carcinogenesis. Male WT and Cry1(-/-) Cry2(-/-) mice (C57Bl/6 background) were chronically exposed to diethylnitrosamine (DEN) at ZT11...
February 22, 2017: International Journal of Cancer. Journal International du Cancer
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