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Fission yeast

Lydia Herzel, Korinna Straube, Karla M Neugebauer
Pre-mRNA splicing is accomplished by the spliceosome, a megadalton complex that assembles de novo on each intron. Because spliceosome assembly and catalysis occur cotranscriptionally, we hypothesized that introns are removed in the order of their transcription in genomes dominated by constitutive splicing. Remarkably little is known about splicing order and the regulatory potential of nascent transcript remodeling by splicing, due to the limitations of existing methods that focus on analysis of mature splicing products (mRNAs) rather than substrates and intermediates...
June 14, 2018: Genome Research
Patricia Richard, Koichi Ogami, Yaqiong Chen, Shuang Feng, James J Moresco, John R Yates, James L Manley
The RNA helicase Mtr4 is a versatile protein that is a crucial component of several distinct RNA surveillance complexes. Here we describe a novel complex that contains Mtr4, but has a role distinct from any of those previously described. We found that Mtr4 association with the human homolog of fission yeast Nrl1, NRDE-2, defines a novel function for Mtr4 in the DNA damage response (DDR) pathway. We provide biochemical evidence that Mtr4 and NRDE-2 are part of the same complex and show that both proteins play a role in the DDR by maintaining low DNA double-strand break levels...
June 14, 2018: RNA Biology
Takahide Hibi, Hokuto Ohtsuka, Takafumi Shimasaki, Shougo Inui, Masatoshi Shibuya, Hideki Tatsukawa, Kei Kanie, Yoshihiko Yamamoto, Hirofumi Aiba
Most antiaging factors or life span extenders are associated with calorie restriction (CR). Very few of these factors function independently of, or additively with, CR. In this study, we focused on tschimganine, a compound that was reported to extend chronological life span (CLS). Although tschimganine led to the extension of CLS, it also inhibited yeast cell growth. We acquired a Schizosaccharomyces pombe mutant with a tolerance for tschimganine due to the gene crm1. The resulting Crm1 protein appears to export the stress-activated protein kinase Sty1 from the nucleus to the cytosol even under stressful conditions...
June 14, 2018: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
Pabitra K Parua, Gregory T Booth, Miriam Sansó, Bradley Benjamin, Jason C Tanny, John T Lis, Robert P Fisher
The end of the RNA polymerase II (Pol II) transcription cycle is strictly regulated to prevent interference between neighbouring genes and to safeguard transcriptome integrity 1 . The accumulation of Pol II downstream of the cleavage and polyadenylation signal can facilitate the recruitment of factors involved in mRNA 3'-end formation and termination 2 , but how this sequence is initiated remains unclear. In a chemical-genetic screen, human protein phosphatase 1 (PP1) isoforms were identified as substrates of positive transcription elongation factor b (P-TEFb), also known as the cyclin-dependent kinase 9 (Cdk9)-cyclin T1 (CycT1) complex 3 ...
June 13, 2018: Nature
Min Lu, Xiangwei He
Distinct chromatin organization features, such as centromeres and heterochromatin domains, are inherited epigenetically. However, the mechanisms that modulate the accuracy of epigenetic inheritance, especially at the individual nucleosome level, are not well understood. Here, using ChIP and next-generation sequencing (ChIP-Seq), we characterized Ccp1, a homolog of the histone chaperone Vps75 in budding yeast that functions in centromere chromatin duplication and heterochromatin maintenance in fission yeast (Schizosaccharomyces pombe)...
June 13, 2018: Journal of Biological Chemistry
Wei-Tong Zhang, En Li, Yan-Kui Guo, Shi-Xia Yu, Zhi-Yuan Wan, Ting Ma, Sha Li, Masa H Sato, Tomoko Hirano, Yan Zhang
Pollen viability depends on dynamic vacuolar changes during pollen development, involving increases and decreases of vacuolar volume through water and osmolite accumulation and vacuolar fission. Mutations in FAB1A-D, the genes encoding phosphatidylinositol 3,5-bisphosphate [PI(3,5)P2]-converting kinases, are male-gametophytic lethal in Arabidopsis thaliana due to defective vacuolar fission after pollen mitosis I (PMI), suggesting a key role of the phospholipid in dynamic vacuolar organization. However, other genetic components that regulate the production of PI(3,5)P2 and its involvement in pollen germination and tube growth are unknown...
June 8, 2018: Plant Physiology
Yang Su, Jie Chen, Ying Huang
Pentatricopeptide repeat (PPR) proteins are major players in mitochondrial and chloroplast RNA metabolism which is essential for normal organellar function. The fission yeast Schizosaccharomyces pombe has ten PPR proteins. We have previously reported that loss of ppr3, ppr4, ppr6, or ppr10 perturbs iron homeostasis leading to accumulation of reactive oxygen species and apoptotic cell death. In the present study, we show that loss of ppr3, ppr4, ppr6, or ppr10 can cause non-sexual flocculation and filamentous growth of cells...
June 6, 2018: FEMS Microbiology Letters
Kosuke Okazaki, Hiroaki Kato, Tetsushi Iida, Kaori Shinmyozu, Jun-Ichi Nakayama, Yota Murakami, Takeshi Urano
BACKGROUND: Heat-shock molecular chaperone proteins (Hsps) promote the loading of small interfering RNA (siRNA) onto RNA interference (RNAi) effector complexes. While the RNAi process is coupled with heterochromatin assembly in several model organisms, it remains unclear whether the Hsps contribute to epigenetic gene regulation. In this study, we used the fission yeast Schizosaccharomyces pombe as a model organism and investigated the roles of Hsp90 and Mas5 (a nucleocytoplasmic type-I Hsp40 protein) in RNAi-dependent heterochromatin assembly...
June 4, 2018: Epigenetics & Chromatin
Hogyu David Seo, Daeyoup Lee
Random mutagenesis of a target gene is commonly used to identify mutations that yield the desired phenotype. Of the methods that may be used to achieve random mutagenesis, error-prone PCR is a convenient and efficient strategy for generating a diverse pool of mutants (i.e., a mutant library). Error-prone PCR is the method of choice when a researcher seeks to mutate a pre-defined region, such as the coding region of a gene while leaving other genomic regions unaffected. After the mutant library is amplified by error-prone PCR, it must be cloned into a suitable plasmid...
May 15, 2018: Journal of Visualized Experiments: JoVE
Xinyun Jing, Bingran Xie, Longxian Chen, Niubing Zhang, Yiyi Jiang, Hang Qin, Hongbing Wang, Pei Hao, Sheng Yang, Xuan Li
In contrast to genome editing, which introduces genetic changes at the DNA level, disrupting or editing gene transcripts provides a distinct approach to perturbing a genetic system, offering benefits complementary to classic genetic approaches. To develop a new toolset for manipulating RNA, we first implemented a member of the type VI CRISPR systems, Cas13a from Leptotrichia shahii (LshCas13a), in Schizosaccharomyces pombe, an important model organism employed by biologists to study key cellular mechanisms conserved from yeast to humans...
May 31, 2018: Nucleic Acids Research
Alba Domènech, José Ayté, Fernando Antunes, Elena Hidalgo
BACKGROUND: Hydrogen peroxide (H2 O2 ) is generated as a by-product of metabolic reactions during oxygen use by aerobic organisms, and can be toxic or participate in signaling processes. Cells, therefore, need to be able to sense and respond to H2 O2 in an appropriate manner. This is often accomplished through thiol switches: Cysteine residues in proteins that can act as sensors, and which are both scarce and finely tuned. Bacteria and eukaryotes use different types of such sensors-either a one-component (OxyR) or two-component (Pap1-Tpx1) redox relay, respectively...
June 1, 2018: BMC Biology
Anna H Klemm, Agneza Bosilj, Matko Gluncˇic, Nenad Pavin, Iva M Tolic
During metaphase, sister chromatids are connected to microtubules extending from the opposite spindle poles via kinetochores to protein complexes on the chromosome. Kinetochores congress to the equatorial plane of the spindle and oscillate around it, with kinesin-8 motors restricting these movements. Yet, the physical mechanism underlying kinetochore movements is unclear. We show that kinetochore movements in the fission yeast Schizosaccharomyces pombe are regulated by kinesin-8-promoted microtubule catastrophe, force-induced rescue, and microtubule dynamic instability...
June 1, 2018: Molecular Biology of the Cell
Touko Niimi, Taro Nakamura
The spindle pole body (SPB) plays a central role in spore plasma membrane formation in addition to its recognized role in microtubule organization. During meiosis, a biomembrane called the forespore membrane (FSM) is newly formed at the SPB. Although several SPB proteins essential for the initiation of FSM formation (meiotic SPB components) have been identified, the molecular mechanism is still unknown. Here, we report the isolation and functional characterization of Dms1 as a component of the SPB. We show that FSM formation does not initiate in dms1Δ cells...
2018: PloS One
Juan Carlos G Cortés, Mariona Ramos, Mami Konomi, Iris Barragán, M Belén Moreno, María Alcaide-Gavilán, Sergio Moreno, Masako Osumi, Pilar Pérez, Juan Carlos Ribas
It is widely accepted in eukaryotes that the cleavage furrow only initiates after mitosis completion. In fission yeast, cytokinesis requires the synthesis of a septum tightly coupled to cleavage furrow ingression. The current cytokinesis model establishes that simultaneous septation and furrow ingression only initiate after spindle breakage and mitosis exit. Thus, this model considers that although Cdk1 is inactivated at early-anaphase, septation onset requires the long elapsed time until mitosis completion and full activation of the Hippo-like SIN pathway...
May 29, 2018: PLoS Genetics
Xun X Bao, Christos Spanos, Tomoko Kojidani, Eric M Lynch, Juri Rappsilber, Yasushi Hiraoka, Tokuko Haraguchi, Kenneth E Sawin
Non-centrosomal microtubule organizing centers (MTOCs) are important for microtubule organization in many cell types. In fission yeast Schizosaccharomyces pombe , the protein Mto1, together with partner protein Mto2 (Mto1/2 complex), recruits the g-tubulin complex to multiple non-centrosomal MTOCs, including the nuclear envelope (NE). Here, we develop a comparative-interactome mass spectrometry approach to determine how Mto1 localizes to the NE. Surprisingly, we find that Mto1, a constitutively cytoplasmic protein, docks at nuclear pore complexes (NPCs), via interaction with exportin Crm1 and cytoplasmic FG-nucleoporin Nup146...
May 29, 2018: ELife
Sojin An, Philipp Koldewey, Jennifer Chik, Lakxmi Subramanian, Uhn-Soo Cho
The Mis18 complex, composed of Mis16, Eic1, and Mis18 in fission yeast, selectively deposits the centromere-specific histone H3 variant, CENP-ACnp1 , at centromeres. How the intact Mis18 holo-complex oligomerizes and how Mis16, a well-known ubiquitous histone H4 chaperone, plays a centromere-specific role in the Mis18 holo-complex, remain unclear. Here, we report the stoichiometry of the intact Mis18 holo-complex as (Mis16)2 :(Eic1)2 :(Mis18)4 using analytical ultracentrifugation. We further determine the crystal structure of Schizosaccharomyces pombe Mis16 in complex with the C-terminal portion of Eic1 (Eic1-CT)...
May 1, 2018: Structure
Tomoyuki Fukuda, Kazuhiro Shiozaki
Target of rapamycin complex 1 (TORC1) is an evolutionarily conserved protein kinase complex, whose activation in response to nutrients suppresses autophagy. In mammalian cells, amino-acid stimuli induce lysosomal translocation and activation of MTORC1 through the RRAG GTPase heterodimer, which is tethered to the surface of lysosomes by the Ragulator complex. Our recent study demonstrated that the fission yeast Schizosaccharomyces pombe also has a Ragulator complex that anchors the Gtr1-Gtr2 Rag GTPase heterodimer to the vacuole, a lysosome-like organelle...
May 25, 2018: Autophagy
Jinyu Wang, Jessica R Eisenstatt, Julien Audry, Kristen Cornelius, Matthew Shaughnessy, Kathleen L Berkner, Kurt W Runge
Heterochromatin domains play important roles in chromosome biology, organismal development and aging, including centromere function, mammalian female X-chromosome inactivation and senescence-associated heterochromatin foci. In the fission yeast Schizosaccharomyces pombe and metazoans, heterochromatin contains histone H3 that is dimethylated at lysine 9. While factors required for heterochromatin have been identified, the dynamics of heterochromatin formation are poorly understood. Telomeres convert adjacent chromatin into heterochromatin...
May 21, 2018: Molecular and Cellular Biology
Hui Li, Fangfu Ye, Jing-Yi Ren, Peng-Ye Wang, Li-Lin Du, Ji-Long Liu
The metabolic enzyme cytidine triphosphate synthase has recently been found to form micron-sized filamentous structures termed cytoophidia, which are evolutionarily conserved across prokaryotes and eukaryotes. The cytoophidium represents a novel type of membraneless organelle and behaves dynamically inside the cell. The question of how cytoophidia transport is mediated, however, remains unanswered. For the first time, we detected in this study the active transport of cytoophidia, taking advantage of the fission yeast Schizosaccharomyces pombe as an excellent model for studying membraneless organelles...
May 21, 2018: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
Ignacio Flor-Parra, Ana Belén Iglesias-Romero, Fred Chang
The organization and number of microtubules (MTs) in a cell depend on the proper regulation of MT nucleation. Currently, the mechanism of nucleation is the most poorly understood aspect of MT dynamics. XMAP215/chTOG/Alp14/Stu2 proteins are MT polymerases that stimulate MT polymerization at MT plus ends by binding and releasing tubulin dimers. Although these proteins also localize to MT organizing centers and have nucleating activity in vitro, it is not yet clear whether these proteins participate in MT nucleation in vivo...
May 10, 2018: Current Biology: CB
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