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https://www.readbyqxmd.com/read/28096973/a-fission-yeast-cell-based-system-for-multidrug-resistant-hiv-1-proteases
#1
Zsigmond Benko, Dong Liang, Ge Li, Robert T Elder, Anindya Sarkar, Jun Takayama, Arun K Ghosh, Richard Y Zhao
BACKGROUND: HIV-1 protease (PR) is an essential enzyme for viral production. Thus, PR inhibitors (PIs) are the most effective class of anti-HIV drugs. However, the main challenge to the successful use of PI drugs in patient treatment is the emergence of multidrug resistant PRs (mdrPRs). This study aimed to develop a fission yeast cell-based system for rapid testing of new PIs that combat mdrPRs. RESULTS: Three mdrPRs were isolated from HIV-infected patients that carried seven (M7PR), ten (M10PR) and eleven (M11PR) PR gene mutations, respectively...
2017: Cell & Bioscience
https://www.readbyqxmd.com/read/28096519/the-nuclear-poly-a-binding-protein-of-mammals-but-not-of-fission-yeast-participates-in-mrna-polyadenylation
#2
Uwe Kühn, Juliane Buschmann, Elmar Wahle
The nuclear poly(A) binding protein (PABPN1) has been suggested, on the basis of biochemical evidence, to play a role in mRNA polyadenylation by strongly increasing the processivity of poly(A) polymerase. While experiments in metazoans have tended to support such a role, the results were not unequivocal, and genetic data show that the S. pombe orthologue of PABPN1, Pab2, is not involved in mRNA polyadenylation. The specific model that PABPN1 increases the rate of poly(A) tail elongation has never been examined in vivo...
January 17, 2017: RNA
https://www.readbyqxmd.com/read/28096402/establishment-of-expression-state-boundaries-by-rif1-and-taz1-in-fission-yeast
#3
Tea Toteva, Bethany Mason, Yutaka Kanoh, Peter Brøgger, Daniel Green, Janne Verhein-Hansen, Hisao Masai, Geneviève Thon
The Shelterin component Rif1 has emerged as a global regulator of the replication-timing program in all eukaryotes examined to date, possibly by modulating the 3D-organization of the genome. In fission yeast a second Shelterin component, Taz1, might share similar functions. Here, we identified unexpected properties for Rif1 and Taz1 by conducting high-throughput genetic screens designed to identify cis- and trans-acting factors capable of creating heterochromatin-euchromatin boundaries in fission yeast. The preponderance of cis-acting elements identified in the screens originated from genomic loci bound by Taz1 and associated with origins of replication whose firing is repressed by Taz1 and Rif1...
January 17, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28093891/multifaceted-effects-of-antimetabolite-and-anticancer-drug-2-deoxyglucose-on-eukaryotic-cancer-models-budding-and-fission-yeast
#4
REVIEW
Akshay Vishwanatha, Cletus Joseph Michael D'Souza
Glycolytic inhibitors are of interest therapeutically as they are effective against cancers that display increased glycolytic rate and mitochondrial defects. 2-Deoxyglucose (2-DG) is one such glycolytic inhibitor and was identified to be a competitive inhibitor of glucose. Studies from past few decades have shown that the mechanism of action of 2-DG is complex involving several metabolic and signaling pathways. Budding yeast Saccharomyces cerevisiae and fission yeast Schizosaccharomyces pombe are two important models for studying metabolism, cell cycle and cell signaling...
January 17, 2017: IUBMB Life
https://www.readbyqxmd.com/read/28089756/glycan-alteration-imparts-cellular-resistance-to-a-membrane-lytic-anticancer-peptide
#5
Ken Ishikawa, Scott H Medina, Joel P Schneider, Amar J S Klar
Although resistance toward small-molecule chemotherapeutics has been well studied, the potential of tumor cells to avoid destruction by membrane-lytic compounds remains unexplored. Anticancer peptides (ACPs) are a class of such agents that disrupt tumor cell membranes through rapid and non-stereospecific mechanisms, encouraging the perception that cellular resistance toward ACPs is unlikely to occur. We demonstrate that eukaryotic cells can, indeed, develop resistance to the model oncolytic peptide SVS-1, which preferentially disrupts the membranes of cancer cells...
January 9, 2017: Cell Chemical Biology
https://www.readbyqxmd.com/read/28087710/targeting-argonaute-to-chromatin
#6
Jered M Wendte, Craig S Pikaard
In many eukaryotes, siRNAs bound to Argonaute proteins guide chromatin-modifying enzymes to complementary loci, resulting in transcriptional gene silencing. Multiple lines of evidence indicate that siRNAs base-pair with longer RNAs produced at target loci, but the possibility that siRNAs base-pair directly with DNA remains an attractive hypothesis. In a recent study, Shimada et al. (pp. 2571-2580) conducted experiments that address these alternative hypotheses, yielding additional evidence that fission yeast siRNA-Argonaute silencing complexes are recruited to target loci exclusively via interactions with nascent transcripts...
December 15, 2016: Genes & Development
https://www.readbyqxmd.com/read/28087675/dna-repair-and-mutations-during-quiescence-in-yeast
#7
Serge Gangloff, Benoit Arcangioli
Life is maintained through alternating phases of cell division and quiescence. The causes and consequences of spontaneous mutations have been extensively explored in proliferating cells, and the major sources include errors of DNA replication and DNA repair. The foremost consequences are genetic variations within a cell population that can lead to heritable diseases and drive evolution. While most of our knowledge on DNA damage response and repair has been gained through cells actively dividing, it remains essential to also understand how DNA damage is metabolized in cells which are not dividing...
January 12, 2017: FEMS Yeast Research
https://www.readbyqxmd.com/read/28069740/checkpoint-independent-regulation-of-origin-firing-by-mrc1-through-interaction-with-hsk1-kinase
#8
Seiji Matsumoto, Yutaka Kanoh, Michie Shimmoto, Motoshi Hayano, Kyosuke Ueda, Rino Fukatsu, Naoko Kakusho, Hisao Masai
Mrc1 is a conserved checkpoint mediator protein that transduces replication-stress signal to downstream effector kinase. Loss of mrc1 checkpoint activity results in aberrant activation of late/dormant origins in the presence of hydroxyurea. Mrc1 was also suggested to regulate orders of early-origin firing in a checkpoint-independent manner, but its mechanism was unknown. Here we identify HBS (Hsk1 Bypass Segment) on Mrc1. ΔHBS does not suppress late/dormant origin firing in the presence of hydroxyurea but causes precocious and enhanced activation of weak early-firing origins during normal S-phase progression, and bypasses the requirement of Hsk1 for growth...
January 9, 2017: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/28065318/in-vitro-reactivation-of-the-cytokinetic-contractile-ring-of-fission-yeast-cells
#9
I Mabuchi, J Kashiwazaki, M Mishra
Cytokinesis is a process by which a mother cell is divided into two daughter cells after chromosome segregation. In both animal and fungal cells, cytokinesis is carried out by the constriction of the contractile ring made up of actin, myosin-II, and other conserved proteins. Detailed genetic and cell biological analysis of cytokinesis has led to the identification of various genes involved in the process of cytokinesis including the cytological description of the process. However, detailed biochemical analysis of the process is lacking...
2017: Methods in Cell Biology
https://www.readbyqxmd.com/read/28065315/nuclear-displacement-and-fluorescence-recovery-after-photobleaching-frap-assays-to-study-division-site-placement-and-cytokinesis-in-fission-yeast
#10
P Ullal, P Bhatia, S G Martin
Cytokinesis is an essential cellular event that completes the cell division cycle. It begins with the assembly of an actomyosin contractile ring that undergoes constriction concomitant with the septum formation to divide the cell in two. Placement of the septum at the right position is important to ensure fidelity of the division process. In fission yeast, the medially placed nucleus is a major spatial cue to position the site of division. In this chapter, we describe a simple synthetic biology-based approach to displace the nucleus and study the consequence on division site positioning...
2017: Methods in Cell Biology
https://www.readbyqxmd.com/read/28057763/the-ultrastructural-organization-of-actin-and-myosin-ii-filaments-in-the-contractile-ring-new-support-for-an-old-model-of-cytokinesis
#11
John H Henson, Casey E Ditzler, Aphnie Germain, Patrick M Irwin, Eric T Vogt, Shucheng Yang, Xufeng Wu, Charles B Shuster
Despite recent advances in our understanding of the components and spatial regulation of the contractile ring (CR), the precise ultrastructure of actin and myosin II within the animal cell CR remains an unanswered question. We employed super-resolution light microscopy and platinum replica transmission electron microscopy (TEM) to determine the structural organization of actin and myosin II in isolated cortical cytoskeletons prepared from dividing sea urchin embryos. 3D structured illumination microscopy (SIM) indicated that within the CR, actin and myosin II filaments were organized into tightly packed linear arrays oriented along the axis of constriction, and restricted to a narrow zone within the furrow...
January 5, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28049830/characterization-of-cytopathic-factors-through-genome-wide-analysis-of-the-zika-viral-proteins-in-fission-yeast
#12
Ge Li, Melissa Poulsen, Csaba Fenyvuesvolgyi, Yoko Yashiroda, Minoru Yoshida, J Marc Simard, Robert C Gallo, Richard Y Zhao
The Zika virus (ZIKV) causes microcephaly and the Guillain-Barré syndrome. Little is known about how ZIKV causes these conditions or which ZIKV viral protein(s) is responsible for the associated ZIKV-induced cytopathic effects, including cell hypertrophy, growth restriction, cell-cycle dysregulation, and cell death. We used fission yeast for the rapid, global functional analysis of the ZIKV genome. All 14 proteins or small peptides were produced under an inducible promoter, and we measured the intracellular localization and the specific effects on ZIKV-associated cytopathic activities of each protein...
January 17, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28049809/electron-microscopy-of-fission-yeast
#13
J Richard McIntosh, Mary K Morphew, Thomas H Giddings
Electron microscopy (EM) can provide images of cells with a spatial resolution that significantly surpasses that available from light microscopy (LM), even with modern methods that give LM "super resolution." However, EM resolution comes with costs in time spent with sample preparation, expense of instrumentation, and concerns regarding sample preparation artifacts. It is therefore important to know the limitations of EM as well as its strengths. Here we describe the most reliable methods for the preservation of fission yeast cells currently available...
January 3, 2017: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/28049779/cryoelectron-microscopy-of-fission-yeast
#14
Mary K Morphew, Thomas H Giddings, J Richard McIntosh
Fission yeast cells can be prepared for electron microscopy (EM) in the frozen-hydrated state. This eliminates the requirement for dehydration and heavy metal staining when preparing samples for EM. As with room temperature imaging, however, the yeast must be sectioned to make them thin enough for transmission of the electron beam. Cutting sections of vitreous ice with a microtome is challenging. An alternative method that uses a focused ion beam to make a thin sample by milling away much of the sample at liquid nitrogen temperatures is under development but is not yet available for routine use...
January 3, 2017: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/28049778/immunolocalization-of-proteins-in-fission-yeast-by-electron-microscopy
#15
Mary K Morphew, Thomas H Giddings, J Richard McIntosh
Electron microscopy (EM) immunolocalization of antigens in fission yeast can be accomplished with cells processed by rapid freezing and freeze-substitution followed by embedding in acrylic or methacrylate resins. Microtome sections of embedded cells are collected onto EM grids. Primary antibodies to the antigen of interest, followed by secondary antibodies conjugated to colloidal gold, are allowed to bind to antigens at the surface of these plastic sections. This type of postembed labeling provides information on antigen localization to a resolution of 10-20 nm, depending on the size of the metal particle used, the form of the antibody (Fab vs...
January 3, 2017: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/28049777/preparing-fission-yeast-for-electron-microscopy
#16
Thomas H Giddings, Mary K Morphew, J Richard McIntosh
Freezing samples while simultaneously subjecting them to a rapid increase in pressure, which inhibits ice crystal formation, is a reliable method for cryofixing fission yeast. The procedure consists simply of harvesting cells and loading them into a high-pressure freezer (HPF), and then operating the device. If equipment for high-pressure freezing is not available, fission yeast can be frozen by plunging a monolayer of cells into a liquid cryogen, usually ethane or propane. Unlike the HPF, where relatively large volumes of cells can be frozen in a single run, plunge freezing requires cells to be dispersed in a layer <20 µm thick...
January 3, 2017: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/28046110/network-centrality-analysis-in-fungi-reveals-complex-regulation-of-lost-and-gained-genes
#17
Jasmin Coulombe-Huntington, Yu Xia
Gene gain and loss shape both proteomes and the networks they form. The increasing availability of closely related sequenced genomes and of genome-wide network data should enable a better understanding of the evolutionary forces driving gene gain, gene loss and evolutionary network rewiring. Using orthology mappings across 23 ascomycete fungi genomes, we identified proteins that were lost, gained or universally conserved across the tree, enabling us to compare genes across all stages of their life-cycle. Based on a collection of genome-wide network and gene expression datasets from baker's yeast, as well as a few from fission yeast, we found that gene loss is more strongly associated with network and expression features of closely related species than that of distant species, consistent with the evolutionary modulation of gene loss propensity through network rewiring...
2017: PloS One
https://www.readbyqxmd.com/read/28041796/a-pp2a-b55-mediated-crosstalk-between-torc1-and-torc2-regulates-the-differentiation-response-in-fission-yeast
#18
Ruth Martín, Marina Portantier, Nathalia Chica, Mari Nyquist-Andersen, Juan Mata, Sandra Lopez-Aviles
Extracellular cues regulate cell fate, and this is mainly achieved through the engagement of specific transcriptional programs. The TORC1 and TORC2 complexes mediate the integration of nutritional cues to cellular behavior, but their interplay is poorly understood. Here, we use fission yeast to investigate how phosphatase activity participates in this interplay during the switch from proliferation to sexual differentiation. We find that loss of PP2A-B55(Pab1) enhances the expression of differentiation-specific genes and leads to premature conjugation...
December 24, 2016: Current Biology: CB
https://www.readbyqxmd.com/read/28032397/dna-base-excision-repair-and-nucleotide-excision-repair-synergistically-contribute-to-survival-of-stationary-phase-cells-of-the-fission-yeast-schizosaccharomyces-pombe
#19
Takanori Senoo, Shinji Kawano, Shogo Ikeda
Defects of genome maintenance may causally contribute to aging. In general, base excision repair (BER) is involved in the repair of subtle base lesions and AP sites, and bulky helix-distorting lesions are restored by nucleotide excision repair (NER). Here, we measured the chronological lifespan (CLS) of BER- and NER-deficient mutants of the fission yeast Schizosaccharomyces pombe, and observed the aging process of cells. The CLS of the nth1 (gene for DNA glycosylase/AP lyase) mutant and the rad16 (a homolog of human XPF) mutant were slightly shorter than that of the wild-type (WT) strain...
December 29, 2016: Cell Biology International
https://www.readbyqxmd.com/read/28031482/involvement-of-fission-yeast-pdc2-in-rna-degradation-and-p-body-function
#20
Chun-Yu Wang, Yi-Ting Wang, Wan-Yi Hsiao, Shao-Win Wang
In this study we identified Pdc2, the fission yeast ortholog of human Pat1b protein, which forms complex with Lsm1-7 and plays a role in coupling deadenylation and decapping. The involvement of Pdc2 in RNA degradation and P-body function were also determined. We found that Pdc2 interacts with Dcp2 and is required for decapping in vivo. Although not absolutely essential for P-bodies assembly, over-expression of Pdc2 enhanced P-body formation even in the absence of Pdc1, the fission yeast functional homologue of human Edc4 protein, indicating that Pdc2 also plays a role in P-body formation...
December 28, 2016: RNA
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