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https://www.readbyqxmd.com/read/29331410/implementing-crispr-cas-technologies-in-conventional-and-non-conventional-yeasts-current-state-and-future-prospects
#1
REVIEW
Hana Raschmanova, Astrid Weninger, Anton Glieder, Karin Kovar, Thomas Vogl
Within five years, the CRISPR-Cas system has emerged as the dominating tool for genome engineering, while also changing the speed and efficiency of metabolic engineering in conventional (Saccharomyces cerevisiae and Schizosaccharomyces pombe) and non-conventional (Yarrowia lipolytica, Pichia pastoris syn. Komagataella phaffii, Kluyveromyces lactis, Candida albicans and C. glabrata) yeasts. Especially in S. cerevisiae, an extensive toolbox of advanced CRISPR-related applications has been established, including crisprTFs and gene drives...
January 10, 2018: Biotechnology Advances
https://www.readbyqxmd.com/read/29330317/trna-production-links-nutrient-conditions-to-the-onset-of-sexual-differentiation-through-the-torc1-pathway
#2
Yoko Otsubo, Tomohiko Matsuo, Akiko Nishimura, Masayuki Yamamoto, Akira Yamashita
Target of rapamycin (TOR) kinase controls cell growth and metabolism in response to nutrient availability. In the fission yeast Schizosaccharomyces pombe, TOR complex 1 (TORC1) promotes vegetative growth and inhibits sexual differentiation in the presence of ample nutrients. Here, we report the isolation and characterization of mutants with similar phenotypes as TORC1 mutants, in that they initiate sexual differentiation even in nutrient-rich conditions. In most mutants identified, TORC1 activity is downregulated and the mutated genes are involved in tRNA expression or modification...
January 12, 2018: EMBO Reports
https://www.readbyqxmd.com/read/29300771/demonstration-of-translation-elongation-factor-3-activity-from-a-non-fungal-species-phytophthora-infestans
#3
Maria K Mateyak, Justyna K Pupek, Alexandra E Garino, McCllelan C Knapp, Sarah F Colmer, Terri Goss Kinzy, Stephen Dunaway
In most eukaryotic organisms, translation elongation requires two highly conserved elongation factors eEF1A and eEF2. Fungal systems are unique in requiring a third factor, the eukaryotic Elongation Factor 3 (eEF3). For decades, eEF3, a ribosome-dependent ATPase, was considered "fungal-specific", however, recent bioinformatics analysis indicates it may be more widely distributed among other unicellular eukaryotes. In order to determine whether divergent eEF3-like proteins from other eukaryotic organisms can provide the essential functions of eEF3 in budding yeast, the eEF3-like proteins from Schizosaccharomyes pombe and an oomycete, Phytophthora infestans, were cloned and expressed in Saccharomyces cerevisiae...
2018: PloS One
https://www.readbyqxmd.com/read/29290612/removal-of-rtf2-from-stalled-replisomes-promotes-maintenance-of-genome-integrity
#4
Molly C Kottemann, Brooke A Conti, Francis P Lach, Agata Smogorzewska
The protection and efficient restart of stalled replication forks is critical for the maintenance of genome integrity. Here, we identify a regulatory pathway that promotes stalled forks recovery from replication stress. We show that the mammalian replisome component C20orf43/RTF2 (homologous to S. pombe Rtf2) must be removed for fork restart to be optimal. We further show that the proteasomal shuttle proteins DDI1 and DDI2 are required for RTF2 removal from stalled forks. Persistence of RTF2 at stalled forks results in fork restart defects, hyperactivation of the DNA damage signal, accumulation of single-stranded DNA (ssDNA), sensitivity to replication drugs, and chromosome instability...
January 4, 2018: Molecular Cell
https://www.readbyqxmd.com/read/29236736/functions-for-fission-yeast-splicing-factors-spslu7-and-spprp18-in-alternative-splice-site-choice-and-stress-specific-regulated-splicing
#5
Geetha Melangath, Titash Sen, Rakesh Kumar, Pushpinder Bawa, Subha Srinivasan, Usha Vijayraghavan
Budding yeast spliceosomal factors ScSlu7 and ScPrp18 interact and mediate intron 3'ss choice during second step pre-mRNA splicing. The fission yeast genome with abundant multi-intronic transcripts, degenerate splice signals and SR proteins is an apt unicellular fungal model to deduce roles for core spliceosomal factors in alternative splice-site choice, intron retention and to study the cellular implications of regulated splicing. From our custom microarray data we deduce a stringent reproducible subset of S...
2017: PloS One
https://www.readbyqxmd.com/read/29235477/nucleotide-and-mal3-dependent-changes-in-fission-yeast-microtubules-suggest-a-structural-plasticity-view-of-dynamics
#6
Ottilie von Loeffelholz, Neil A Venables, Douglas Robert Drummond, Miho Katsuki, Robert Cross, Carolyn A Moores
Using cryo-electron microscopy, we characterize the architecture of microtubules assembled from Schizosaccharomyces pombe tubulin, in the presence and absence of their regulatory partner Mal3. Cryo-electron tomography reveals that microtubules assembled from S. pombe tubulin have predominantly B-lattice interprotofilament contacts, with protofilaments skewed around the microtubule axis. Copolymerization with Mal3 favors 13 protofilament microtubules with reduced protofilament skew, indicating that Mal3 adjusts interprotofilament interfaces...
December 13, 2017: Nature Communications
https://www.readbyqxmd.com/read/29234668/a-genome-wide-screen-for-fty720-sensitive-mutants-reveals-genes-required-for-ros-homeostasis
#7
Kanako Hagihara, Kanako Kinoshita, Kouki Ishida, Shihomi Hojo, Yoshinori Kameoka, Ryosuke Satoh, Teruaki Takasaki, Reiko Sugiura
Fingolimod hydrochloride (FTY720), a sphingosine-1-phosphate (S1P) analogue, is an approved immune modulator for the treatment of multiple sclerosis (MS). Notably, in addition to its well-known mode of action as an S1P modulator, accumulating evidence suggests that FTY720 induces apoptosis in various cancer cells via reactive oxygen species (ROS) generation. Although the involvement of multiple signaling molecules, such as JNK (Jun N-terminal kinase), Akt (alpha serine/threonine-protein kinase) and Sphk has been reported, the exact mechanisms how FTY720 induces cell growth inhibition and the functional relationship between FTY720 and these signaling pathways remain elusive...
November 27, 2017: Microbial Cell
https://www.readbyqxmd.com/read/29232693/host-factors-that-promote-retrotransposon-integration-are-similar-in-distantly-related-eukaryotes
#8
Sudhir Kumar Rai, Maya Sangesland, Michael Lee, Caroline Esnault, Yujin Cui, Atreyi Ghatak Chatterjee, Henry L Levin
Retroviruses and Long Terminal Repeat (LTR)-retrotransposons have distinct patterns of integration sites. The oncogenic potential of retrovirus-based vectors used in gene therapy is dependent on the selection of integration sites associated with promoters. The LTR-retrotransposon Tf1 of Schizosaccharomyces pombe is studied as a model for oncogenic retroviruses because it integrates into the promoters of stress response genes. Although integrases (INs) encoded by retroviruses and LTR-retrotransposons are responsible for catalyzing the insertion of cDNA into the host genome, it is thought that distinct host factors are required for the efficiency and specificity of integration...
December 12, 2017: PLoS Genetics
https://www.readbyqxmd.com/read/29220656/mpk1-slt2-links-multiple-stress-responses-with-gene-expression-in-budding-yeast-by-phosphorylating-tyr1-of-the-rnap-ii-ctd
#9
Nathan Yurko, Xiaochuan Liu, Takashi Yamazaki, Mainul Hoque, Bin Tian, James L Manley
The RNA polymerase II largest subunit C-terminal domain consists of repeated YSPTSPS heptapeptides. The role of tyrosine-1 (Tyr1) remains incompletely understood, as, for example, mutating all Tyr1 residues to Phe (Y1F) is lethal in vertebrates but a related mutant has only a mild phenotype in S. pombe. Here we show that Y1F substitution in budding yeast resulted in a strong slow-growth phenotype. The Y1F strain was also hypersensitive to several different cellular stresses that involve MAP kinase signaling...
December 7, 2017: Molecular Cell
https://www.readbyqxmd.com/read/29205883/fission-yeast-myo2-molecular-organization-and-diffusion-in-the-cytoplasm
#10
Janice E Friend, Wasim A Sayyad, Rajesh Arasada, Chad D McCormick, John E Heuser, Thomas D Pollard
Myosin-II is required for the assembly and constriction of cytokinetic contractile rings in fungi and animals. We used electron microscopy, fluorescence recovery after photobleaching (FRAP), and fluorescence correlation spectroscopy (FCS) to characterize the physical properties of Myo2 from fission yeast Schizosaccharomyces pombe. By electron microscopy, Myo2 has two heads and a coiled-coiled tail like myosin-II from other species. The first 65 nm of the tail is a stiff rod, followed by a flexible, less-ordered region up to 30 nm long...
December 4, 2017: Cytoskeleton
https://www.readbyqxmd.com/read/29195389/multiplexed-fluctuation-dissipation-theorem-calibration-of-optical-tweezers-inside-living-cells
#11
Hao Yan, Jessica F Johnston, Sidney B Cahn, Megan C King, Simon G J Mochrie
In order to apply optical tweezers-based force measurements within an uncharacterized viscoelastic medium such as the cytoplasm of a living cell, a quantitative calibration method that may be applied in this complex environment is needed. We describe an improved version of the fluctuation-dissipation-theorem calibration method, which has been developed to perform in situ calibration in viscoelastic media without prior knowledge of the trapped object. Using this calibration procedure, it is possible to extract values of the medium's viscoelastic moduli as well as the force constant describing the optical trap...
November 2017: Review of Scientific Instruments
https://www.readbyqxmd.com/read/29191091/schizosaccharomyces-japonicus-has-low-levels-of-coq10-synthesis-respiration-deficiency-and-efficient-ethanol-production
#12
Tomohiro Kaino, Kai Tonoko, Shiomi Mochizuki, Yuriko Takashima, Makoto Kawamukai
Coenzyme Q (CoQ) is essential for mitochondrial respiration and as a cofactor for sulfide quinone reductase. Schizosaccharomyces pombe produces a human-type CoQ10. Here, we analyzed CoQ in other fission yeast species. S. cryophilus and S. octosporus produce CoQ9. S. japonicus produces low levels of CoQ10, although all necessary genes for CoQ synthesis have been identified in its genome. We expressed three genes (dps1, dlp1, and ppt1) for CoQ synthesis from S. japonicus in the corresponding S. pombe mutants, and confirmed that they were functional...
December 1, 2017: Bioscience, Biotechnology, and Biochemistry
https://www.readbyqxmd.com/read/29188185/metabolic-engineering-of-schizosaccharomyces-pombe-via-crispr-cas9-genome-editing-for-lactic-acid-production-from-glucose-and-cellobiose
#13
Aiko Ozaki, Rie Konishi, Chisako Otomo, Mayumi Kishida, Seiya Takayama, Takuya Matsumoto, Tsutomu Tanaka, Akihiko Kondo
Modification of the Schizosaccharomyces pombe genome is often laborious, time consuming due to the lower efficiency of homologous recombination. Here, we constructed metabolically engineered S. pombe strains using a CRISPR-Cas9 system and also demonstrated D-lactic acid (D-LA) production from glucose and cellobiose. Genes encoding two separate pyruvate decarboxylases (PDCs), an L-lactic acid dehydrogenase (L-LDH), and a minor alcohol dehydrogenase (SPBC337.11) were disrupted, thereby attenuating ethanol production...
December 2017: Metabolic Engineering Communications
https://www.readbyqxmd.com/read/29187422/dependency-of-heterochromatin-domains-on-replication-factors
#14
Leonie Johanna Jahn, Bethany Mason, Peter Brøgger, Tea Toteva, Dennis Kim Nielsen, Genevieve Thon
Chromatin structure regulates both genome expression and dynamics in eukaryotes where large heterochromatic regions are epigenetically silenced through the methylation of histone H3K9, histone deacetylation, and assembly of repressive complexes. Previous genetic screens with the fission yeast Schizosaccharomyces pombe have led to the identification of key enzymatic activities and structural constituents of heterochromatin. We report here on additional factors discovered by screening a library of deletion mutants for silencing defects at the edge of a heterochromatic domain bound by its natural boundary - the IR-R+ element - or by ectopic boundaries...
November 29, 2017: G3: Genes—Genomes—Genetics
https://www.readbyqxmd.com/read/29183750/pheromone-inducible-expression-vectors-for-fission-yeast-schizosaccharomyces-pombe
#15
Stefan Hennig, Nadine Hornauer, Gerhard Rödel, Kai Ostermann
The fission yeast Schizosaccharomyces pombe is an attractive host for heterologous gene expression. However, expression systems for industrially viable large-scale fermentations are scarce. Several inducible expression vectors for S. pombe have been reported, with the strong thiamine-repressible nmt1+ promoter or derivatives thereof most commonly employed. Previously, the promoter regions of the genes sxa2+ and rep1+ were utilized to couple pheromone signaling to the expression of reporter genes for quantitative assessment of the cellular response to mating pheromones...
November 25, 2017: Plasmid
https://www.readbyqxmd.com/read/29180432/phosphorylation-of-cenp-c-by-aurora-b-facilitates-kinetochore-attachment-error-correction-in-mitosis
#16
Xing Zhou, Fan Zheng, Chengliang Wang, Minhao Wu, Xiaozhen Zhang, Qian Wang, Xuebiao Yao, Chuanhai Fu, Xuan Zhang, Jianye Zang
Kinetochores are superprotein complexes that orchestrate chromosome segregation via a dynamic interaction with spindle microtubules. A physical connection between CENP-C and the Mis12-Ndc80-Knl1 (KMN) protein network is an important pathway that is used to assemble kinetochores on CENP-A nucleosomes. Multiple outer kinetochore components are phosphorylated by Aurora B kinase to activate the spindle assembly checkpoint (SAC) and to ensure accurate chromosome segregation. However, it is unknown whether Aurora B can phosphorylate inner kinetochore components to facilitate proper mitotic chromosome segregation...
November 27, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29168508/structural-basis-for-the-initiation-of-eukaryotic-transcription-coupled-dna-repair
#17
Jun Xu, Indrajit Lahiri, Wei Wang, Adam Wier, Michael A Cianfrocco, Jenny Chong, Alissa A Hare, Peter B Dervan, Frank DiMaio, Andres E Leschziner, Dong Wang
Eukaryotic transcription-coupled repair (TCR) is an important and well-conserved sub-pathway of nucleotide excision repair that preferentially removes DNA lesions from the template strand that block translocation of RNA polymerase II (Pol II). Cockayne syndrome group B (CSB, also known as ERCC6) protein in humans (or its yeast orthologues, Rad26 in Saccharomyces cerevisiae and Rhp26 in Schizosaccharomyces pombe) is among the first proteins to be recruited to the lesion-arrested Pol II during the initiation of eukaryotic TCR...
November 22, 2017: Nature
https://www.readbyqxmd.com/read/29167352/two-spatially-distinct-kinesin-14-pkl1-and-klp2-generate-collaborative-inward-forces-against-kinesin-5-cut7-in-s-pombe
#18
Masashi Yukawa, Yusuke Yamada, Tomoaki Yamauchi, Takashi Toda
Kinesin motors play central roles in bipolar spindle assembly. In many eukaryotes, spindle pole separation is driven by Kinesin-5 that generates outward force. This outward force is balanced by antagonistic inward force elicited by Kinesin-14 and/or Dynein. In fission yeast, two Kinesin-14s, Pkl1 and Klp2, play an opposing role against Kinesin-5/Cut7. However, how these two Kinesin-14s coordinate individual activities remains elusive. Here we show that while deletion of either pkl1 or klp2 rescues temperature sensitive cut7 mutants, only pkl1 deletion can bypass the lethality caused by cut7 deletion...
November 22, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29160296/structure-of-the-fission-yeast-s-pombe-telomeric-tpz1-poz1-rap1-complex
#19
Jing Xue, Hongwen Chen, Jian Wu, Miho Takeuchi, Haruna Inoue, Yanmei Liu, Hong Sun, Yong Chen, Junko Kanoh, Ming Lei
Telomeric shelterin complex caps chromosome ends and plays a crucial role in telomere maintenance and protection. In the fission yeast Schizosaccharomyces pombe, shelterin is composed of telomeric single- and double-stranded DNA-binding protein subcomplexes Pot1-Tpz1 and Taz1-Rap1, which are bridged by their interacting protein Poz1. However, the structure of Poz1 and how Poz1 functions as an interaction hub in the shelterin complex remain unclear. Here we report the crystal structure of Poz1 in complex with Poz1-binding motifs of Tpz1 and Rap1...
November 21, 2017: Cell Research
https://www.readbyqxmd.com/read/29155976/description-of-a-mass-poisoning-in-a-rural-district-in-mozambique-the-first-documented-bongkrekic-acid-poisoning-in-africa
#20
Eduardo Samo Gudo, Kyla Cook, Amelia M Kasper, Alfredo Vergara, Cristolde Salomão, Fernanda Oliveira, Hamida Ismael, Cristovão Saeze, Carla Mosse, Quinhas Fernandes, Sofia Omar Viegas, Cynthia S Baltazar, Timothy J Doyle, Ellen Yard, Alaina Steck, Mayda Serret, Travis M Falconer, Sara E Kern, Jennifer L Brzezinski, James A Turner, Brian L Boyd, Ilesh V Jani
Background: On January 9, 2015, in a rural town in Mozambique, over 230 people became sick and 75 died from an illness linked to drinking pombe, a traditional alcoholic beverage. Methods: An investigation was conducted to identify cases and determine the cause of the outbreak. A case was defined as any resident of Chitima who developed any new or unexplained neurologic, gastrointestinal, or cardiovascular symptom from January 9 at 6:00 a.m. through January 12. We conducted medical record reviews; healthcare worker and community surveys; anthropological and toxicological investigations of local medicinal plants and commercial pesticides; and laboratory testing of the suspect and control pombe...
November 16, 2017: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America
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