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https://www.readbyqxmd.com/read/28432181/dual-nature-of-pseudouridylation-in-u2-snrna-pus1p-dependent-and-pus1p-independent-activities-in-yeasts-and-higher-eukaryotes
#1
Svetlana Deryusheva, Joseph G Gall
The pseudouridine at position 43 in vertebrate U2 snRNA is one of the most conserved posttranscriptional modifications of spliceosomal snRNAs; the equivalent position is pseudouridylated in U2 snRNAs in different phyla including fungi, insects, and worms. Pseudouridine synthase Pus1p acts alone on U2 snRNA to form this pseudouridine in yeast Saccharomyces cerevisiae and mouse. Furthermore, in S. cerevisiae Pus1p is the only pseudouridine synthase for this position. Using an in vivo yeast cell system we tested enzymatic activity of Pus1p from the fission yeast Schizosaccharomyces pombe, the worm Caenorhabditis elegans, the fruit fly Drosophila melanogaster and the frog Xenopus tropicalis...
April 21, 2017: RNA
https://www.readbyqxmd.com/read/28426144/a-multiplex-culture-system-for-the-long-term-growth-of-fission-yeast-cells
#2
Celine Callens, Nelson C Coelho, Aaron W Miller, Maria Rosa Domino Sananes, Maitreya J Dunham, Matthieu Denoual, Damien Coudreuse
Maintenance of long-term cultures of yeast cells is central to a broad range of investigations, from metabolic studies to laboratory evolution assays. However, repeated dilutions of batch cultures lead to variations in medium composition, with implications for cell physiology. In S. cerevisiae, powerful miniaturized chemostat setups, or ministat arrays, have been shown to allow for constant dilution of multiple independent cultures. Here we set out to adapt these arrays for continuous culture of a morphologically and physiologically distinct yeast, the fission yeast Schizosaccharomyces pombe, with the goal of maintaining constant population density over time...
April 20, 2017: Yeast
https://www.readbyqxmd.com/read/28424666/napa-mediates-a-redox-regulation-of-the-antioxidant-response-carbon-utilization-and-development-in-aspergillus-nidulans
#3
Ariann E Mendoza-Martínez, Fernando Lara-Rojas, Olivia Sánchez, Jesús Aguirre
The redox-regulated transcription factors (TFs) of the bZIP AP1 family, such as yeast Yap1 and fission yeast Pap1, are activated by peroxiredoxin proteins (Prxs) to regulate the antioxidant response. Previously, Aspergillus nidulans mutants lacking the Yap1 ortholog NapA have been characterized as sensitive to H2O2 and menadione. Here we study NapA roles in relation to TFs SrrA and AtfA, also involved in oxidant detoxification, showing that these TFs play different roles in oxidative stress resistance, catalase gene regulation and development, during A...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28423198/an-estradiol-inducible-promoter-enables-fast-graduated-control-of-gene-expression-in-fission-yeast
#4
Makoto J Ohira, David G Hendrickson, R Scott McIsaac, Nicholas Rhind
The fission yeast Schizosaccharomyces pombe lacks a diverse toolkit of inducible promoters for experimental manipulation. Available inducible promoters suffer from slow induction kinetics, limited control of expression levels and/or a requirement for defined growth medium. In particular, no S. pombe inducible promoter systems exhibit a linear dose response, which would allow expression to be tuned to specific levels. We have adapted a fast, orthogonal promoter system with a large dynamic range and a linear dose response, based on β-estradiol-regulated function of the human estrogen receptor, for use in S...
April 19, 2017: Yeast
https://www.readbyqxmd.com/read/28410370/a-systematic-screen-for-morphological-abnormalities-during-fission-yeast-sexual-reproduction-identifies-a-mechanism-of-actin-aster-formation-for-cell-fusion
#5
Omaya Dudin, Laura Merlini, Felipe O Bendezú, Raphaël Groux, Vincent Vincenzetti, Sophie G Martin
In non-motile fungi, sexual reproduction relies on strong morphogenetic changes in response to pheromone signaling. We report here on a systematic screen for morphological abnormalities of the mating process in fission yeast Schizosaccharomyces pombe. We derived a homothallic (self-fertile) collection of viable deletions, which, upon visual screening, revealed a plethora of phenotypes affecting all stages of the mating process, including cell polarization, cell fusion and sporulation. Cell fusion relies on the formation of the fusion focus, an aster-like F-actin structure that is marked by strong local accumulation of the myosin V Myo52, which concentrates secretion at the fusion site...
April 14, 2017: PLoS Genetics
https://www.readbyqxmd.com/read/28396503/correlation-of-meiotic-dsb-formation-and-transcription-initiation-around-fission-yeast-recombination-hotspots
#6
Shintaro Yamada, Mika Okamura, Arisa Oda, Hiroshi Murakami, Kunihiro Ohta, Takatomi Yamada
Meiotic homologous recombination, a critical event for ensuring faithful chromosome segregation and creating genetic diversity, is initiated by programed DNA double strand breaks (DSBs) formed at recombination hotspots. Meiotic DSB formation is likely to be influenced by other DNA-templated processes including transcription, but how DSB formation and transcription interact with each other has not been understood well. In this study, we used fission yeast to investigate a possible interplay of these two events...
April 10, 2017: Genetics
https://www.readbyqxmd.com/read/28388826/sulfur-restriction-extends-fission-yeast-chronological-lifespan-through-ecl1-family-genes-by-downregulation-of-ribosome
#7
Hokuto Ohtsuka, Masahiro Takinami, Takafumi Shimasaki, Takahide Hibi, Hiroshi Murakami, Hirofumi Aiba
Nutritional restrictions such as calorie restrictions are known to increase the lifespan of various organisms. Here we found that a restriction of sulfur extended the chronological lifespan (CLS) of the fission yeast Schizosaccharomyces pombe. The restriction decreased cellular size, RNA content, and ribosomal proteins and increased sporulation rate. These responses depended on Ecl1 family genes, the overexpression of which results in the extension of CLS. We also showed that the Zip1 transcription factor results in the sulfur restriction-dependent expression of the ecl1(+) gene...
April 7, 2017: Molecular Microbiology
https://www.readbyqxmd.com/read/28382430/dna-topoisomerase-ii-modulates-acetyl-regulation-of-cohesin-mediated-chromosome-dynamics
#8
Su-Jiun Lin, Matthew J O'Connell
Cohesin is one of three multi-protein structural maintenance of chromosome (SMC) complexes that regulate eukaryotic chromosome dynamics. It forms a ring-shaped structure that embraces sister chromatids through interphase to promote their pairing. In preparation for mitosis, most cohesin is stripped from the chromosome arms in prophase by a poorly defined process that is associated with cohesin phosphorylation. In the fission yeast Schizosaccharomyces pombe this prophase pathway is dependent on the cohesin-related Smc5/6 complex, and this requirement is heightened in Smc5/6 hypomorphs by DNA damage, replication stress and Topoisomerase II (Top2) dysfunction...
April 5, 2017: Current Genetics
https://www.readbyqxmd.com/read/28377506/heme-deficiency-sensitizes-yeast-cells-to-oxidative-stress-induced-by-hydroxyurea
#9
Amanpreet Singh, Yong-Jie Xu
Hydroxyurea (HU) has a long history of clinical and scientific use as an antiviral, antibacterial, and antitumor agent. It inhibits ribonucleotide reductase and reversibly arrests cells in S phase. However, high concentrations or prolonged treatment with low doses of HU can cause cell lethality. Although the cytotoxicity of HU may significantly contribute to its therapeutic effects, the underlying mechanisms remain poorly understood. We have previously shown that HU can induce cytokinesis arrest in the erg11-1 mutant of fission yeast, which has a partial defect in the biosynthesis of fungal membrane sterol ergosterol (1)...
April 4, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28373489/preparation-of-total-rna-from-fission-yeast
#10
Jürg Bähler, Jo Ann Wise
Treatment with hot phenol breaks open fission yeast cells and begins to strip away bound proteins from RNA. Deproteinization is completed by multiple extractions with chloroform/isoamyl alcohol and separation of the aqueous and organic phases using MaXtract gel, an inert material that acts as a physical barrier between the phases. The final step is concentration of the RNA by ethanol precipitation. The protocol can be used to prepare RNA from several cultures grown in parallel, but it is important not to process too many samples at once because delays can be detrimental to RNA quality...
April 3, 2017: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/28367989/the-conserved-protein-seb1-drives-transcription-termination-by-binding-rna-polymerase-ii-and-nascent-rna
#11
Sina Wittmann, Max Renner, Beth R Watts, Oliver Adams, Miles Huseyin, Carlo Baejen, Kamel El Omari, Cornelia Kilchert, Dong-Hyuk Heo, Tea Kecman, Patrick Cramer, Jonathan M Grimes, Lidia Vasiljeva
Termination of RNA polymerase II (Pol II) transcription is an important step in the transcription cycle, which involves the dislodgement of polymerase from DNA, leading to release of a functional transcript. Recent studies have identified the key players required for this process and showed that a common feature of these proteins is a conserved domain that interacts with the phosphorylated C-terminus of Pol II (CTD-interacting domain, CID). However, the mechanism by which transcription termination is achieved is not understood...
April 3, 2017: Nature Communications
https://www.readbyqxmd.com/read/28366744/fission-yeast-apc15-stabilizes-mcc-cdc20-apc-c-complexes-ensuring-efficient-cdc20-ubiquitination-and-checkpoint-arrest
#12
Karen M May, Flora Paldi, Kevin G Hardwick
During mitosis, cells must segregate the replicated copies of their genome to their daughter cells with extremely high fidelity. Segregation errors lead to an abnormal chromosome number (aneuploidy), which typically results in disease or cell death [1]. Chromosome segregation and anaphase onset are initiated through the action of the multi-subunit E3 ubiquitin ligase known as the anaphase-promoting complex or cyclosome (APC/C [2]). The APC/C is inhibited by the spindle checkpoint in the presence of kinetochore attachment defects [3, 4]...
March 28, 2017: Current Biology: CB
https://www.readbyqxmd.com/read/28366743/different-functionality-of-cdc20-binding-sites-within-the-mitotic-checkpoint-complex
#13
Katharina Sewart, Silke Hauf
The mitotic checkpoint is a cellular safeguard that prevents chromosome missegregation in eukaryotic cells [1, 2]. Suboptimal functioning may foster chromosome missegregation in cancer cells [3]. Checkpoint signaling produces the "mitotic checkpoint complex" (MCC), which prevents anaphase by targeting Cdc20, the activator of the anaphase-promoting complex/cyclosome (APC/C). Recent biochemical and structural studies revealed that the human MCC binds two Cdc20 molecules, one (Cdc20(M)) through well-characterized, cooperative binding to Mad2 and Mad3/BubR1 (forming the "core MCC") and the other one (Cdc20(A)) through additional binding sequences in Mad3/BubR1 [4-6]...
March 28, 2017: Current Biology: CB
https://www.readbyqxmd.com/read/28357352/chromatin-binding-and-silencing-two-roles-of-the-same-protein-lem2
#14
COMMENT
Ramón Ramos Barrales, Sigurd Braun
Transcriptionally repressed chromatin localizes to specific areas within the eukaryotic nucleus and is often found at the nuclear periphery, which is thought to provide a specialized compartment for gene silencing. However, the molecular mechanisms that establish this spatial chromatin organization are still poorly understood. In our recent work (Barrales et al. 2016), we identified the nuclear envelope protein Lem2, a homolog of metazoan lamin-associated proteins (LAPs), as a relevant factor for heterochromatin silencing and perinuclear localization in the fission yeast Schizosaccharomyces pombe...
April 4, 2016: Microbial Cell
https://www.readbyqxmd.com/read/28352647/single-step-marker-switching-in-schizosaccharomyces-pombe-using-a-lithium-acetate-transformation-protocol
#15
Simon D Brown, Alexander Lorenz
The ability to utilize different selectable markers for tagging or mutating multiple genes in Schizosaccharomyces pombe is hampered by the historical use of only two selectable markers, ura4(+) and kanMX6; the latter conferring resistance to the antibiotic G418 (geneticin). More markers have been described recently, but introducing these into yeast cells often requires strain construction from scratch. To overcome this problem we and other groups have created transformation cassettes with flanking homologies to ura4(+) and kanMX6 which enable an efficient and time-saving way to exchange markers in existing mutated or tagged fission yeast strains...
December 20, 2016: Bio-protocol
https://www.readbyqxmd.com/read/28350990/the-putative-drp1-inhibitor-mdivi-1-is-a-reversible-mitochondrial-complex-i-inhibitor-that-modulates-reactive-oxygen-species
#16
Evan A Bordt, Pascaline Clerc, Brian A Roelofs, Andrew J Saladino, László Tretter, Vera Adam-Vizi, Edward Cherok, Ahmed Khalil, Nagendra Yadava, Shealinna X Ge, T Chase Francis, Nolan W Kennedy, Lora K Picton, Tanya Kumar, Sruti Uppuluri, Alexandrea M Miller, Kie Itoh, Mariusz Karbowski, Hiromi Sesaki, R Blake Hill, Brian M Polster
Mitochondrial fission mediated by the GTPase dynamin-related protein 1 (Drp1) is an attractive drug target in numerous maladies that range from heart disease to neurodegenerative disorders. The compound mdivi-1 is widely reported to inhibit Drp1-dependent fission, elongate mitochondria, and mitigate brain injury. Here, we show that mdivi-1 reversibly inhibits mitochondrial complex I-dependent O2 consumption and reverse electron transfer-mediated reactive oxygen species (ROS) production at concentrations (e...
March 27, 2017: Developmental Cell
https://www.readbyqxmd.com/read/28349391/ribosome-profiling-for-the-analysis-of-translation-during-yeast-meiosis
#17
Caia Duncan, Juan Mata
Ribosome profiling provides a genome-wide view of translation with unprecedented resolution. Application of this approach to fission and budding yeast revealed widespread regulation of translational efficiency, translation of short open reading frames on unannotated transcripts, and frequent translation of open reading frames in 5' leader sequences. We present here a detailed protocol for the application of ribosome profiling to meiotic fission yeast cells, although the approach should be easily adapted to budding yeast...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28349389/quantitative-genome-wide-measurements-of-meiotic-dna-double-strand-breaks-and-protein-binding-in-s-pombe
#18
Randy W Hyppa, Kyle R Fowler, Gerald R Smith
The fission yeast Schizosaccharomyces pombe is especially well suited for studying meiosis in molecular detail. Experiments with S. pombe strains that undergo a nearly synchronous meiosis-at variable temperatures-have elucidated the mechanisms of meiotic progression and the proteins that are involved. For example, studies focused on the initiation of meiotic recombination by programmed DNA double-strand breaks (DSBs) have proven exceptionally informative. In meiosis, some regions of DNA have more frequent DSBs than the surrounding regions...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28348222/form-and-function-of-topologically-associating-genomic-domains-in-budding-yeast
#19
Umut Eser, Devon Chandler-Brown, Ferhat Ay, Aaron F Straight, Zhijun Duan, William Stafford Noble, Jan M Skotheim
The genome of metazoan cells is organized into topologically associating domains (TADs) that have similar histone modifications, transcription level, and DNA replication timing. Although similar structures appear to be conserved in fission yeast, computational modeling and analysis of high-throughput chromosome conformation capture (Hi-C) data have been used to argue that the small, highly constrained budding yeast chromosomes could not have these structures. In contrast, herein we analyze Hi-C data for budding yeast and identify 200-kb scale TADs, whose boundaries are enriched for transcriptional activity...
April 11, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28345120/fission-yeast-srm1-is-involved-in-stress-response-and-cell-cycle
#20
Aslıhan Örs Gevrekci
Polyamines are well-conserved, multifunctional polycations that contribute to a number of processes in the cells such as cell cycle, apoptosis, stress response, and gene expression. Therefore, polyamine levels should be kept under strict regulation by specific polyamine transporters and polyamine synthases. In this study, the aim is to experimentally characterize a predicted spermidine synthase gene srm1, which was identified upon sequence similarity, in fission yeast Schizosaccharomyces pombe. In an attempt to understand the role of this gene in cell cycle and stress response, deletion mutant of srm1 was generated and analyzed in terms of cell cycle regulation and environmental stress response...
March 27, 2017: Current Microbiology
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