Schizosaccharomyces Pombe

14,000,000 Leading Edge Experts on the ideXlab platform

Scan Science and Technology

Contact Leading Edge Experts & Companies

Scan Science and Technology

Contact Leading Edge Experts & Companies

The Experts below are selected from a list of 20889 Experts worldwide ranked by ideXlab platform

Yasushi Hiraoka - One of the best experts on this subject based on the ideXlab platform.

John Davey - One of the best experts on this subject based on the ideXlab platform.

  • the sxa2 dependent inactivation of the p factor mating pheromone in the fission yeast Schizosaccharomyces Pombe
    Molecular Microbiology, 1996
    Co-Authors: Graham Ladds, Olaf Nielsen, Erik Michael Rasmussen, Tom W Young, John Davey
    Abstract:

    Summary Haploid cells of the fission yeast Schizosaccharomyces Pombe exist in one of two mating types, referred to as M and P. Conjugation occurs between cells of opposite mating type and is controlled by the reciprocal action of diffusible pheromones. Loss of function of the sxa2 gene in M cells causes hypersensitivity to the P-factor mating pheromone and a reduction in mating efficiency. Here we demonstrate the secretion of an sxa2-dependent carboxypeptidase that inactivates P-factor by removal of the C-terminal leucine residue.

  • chemical synthesis of the m factor mating pheromone from Schizosaccharomyces Pombe
    Yeast, 1994
    Co-Authors: Shuhua Wang, John Davey, Chubiao Xue, Olaf Nielsen, Fred Naider
    Abstract:

    Conjugation in the fission yeast Schizosaccharomyces Pombe is controlled by the reciprocal action of mating pheromones. We recently showed that M-factor, the pheromone released by cells of the cellular mating type Minus, is a nonapeptide in which the C-terminal cysteine residue is carboxyl-methylated and S-alkylated, probably with a farnesyl residue (Davey, 1992): Tyr-Thr-Pro-Lys-Val-Pro-Tyr-Met-Cys(S-farnesyl)- OCH3. Here we describe the chemical synthesis of this modified peptide and show that it exhibits all of the properties of the native pheromone. These results confirm the structure of the M-factor while the production of relatively large amounts of pure pheromone will be invaluable for studying the mating response in this yeast.

Antony M Carr - One of the best experts on this subject based on the ideXlab platform.

  • transformation of Schizosaccharomyces Pombe lithium acetate dimethyl sulfoxide procedure
    CSH Protocols, 2016
    Co-Authors: Johanne M Murray, Adam T Watson, Antony M Carr
    Abstract:

    Transformation of Schizosaccharomyces Pombe with DNA requires the conditioning of cells to promote DNA uptake followed by cell growth under conditions that select and maintain the plasmid or integration event. The three main methodologies are electroporation, treatment with lithium cations, and transformation of protoplasts. The lithium acetate method described here is widely used because it is simple and reliable.

  • Postreplication Repair and PCNA Modification in Schizosaccharomyces Pombe
    Molecular biology of the cell, 2006
    Co-Authors: Jonathan Frampton, Antony M Carr, Kanji Furuya, Anja Irmisch, Catherine M. Green, Andrea Neiss, Michelle Trickey, Helle D. Ulrich, Felicity Z. Watts, Alan R. Lehmann
    Abstract:

    Ubiquitination of proliferating cell nuclear antigen (PCNA) plays a crucial role in regulating replication past DNA damage in eukaryotes, but the detailed mechanisms appear to vary in different organisms. We have examined the modification of PCNA in Schizosaccharomyces Pombe. We find that, in response to UV irradiation, PCNA is mono- and poly-ubiquitinated in a manner similar to that in Saccharomyces cerevisiae. However in undamaged Schizosaccharomyces Pombe cells, PCNA is ubiquitinated in S phase, whereas in S. cerevisiae it is sumoylated. Furthermore we find that, unlike in S. cerevisiae, mutants defective in ubiquitination of PCNA are also sensitive to ionizing radiation, and PCNA is ubiquitinated after exposure of cells to ionizing radiation, in a manner similar to the response to UV-irradiation. We show that PCNA modification and cell cycle checkpoints represent two independent signals in response to DNA damage. Finally, we unexpectedly find that PCNA is ubiquitinated in response to DNA damage when cells are arrested in G2.

  • the Schizosaccharomyces Pombe rad3 checkpoint gene
    The EMBO Journal, 1996
    Co-Authors: Nicola J Bentley, D A Holtzman, G Flaggs, K S Keegan, A Demaggio, J C Ford, M Hoekstra, Antony M Carr
    Abstract:

    The rad3 gene of Schizosaccharomyces Pombe is required for checkpoint pathways that respond to DNA damage and replication blocks. We report the complete rad3 gene sequence and show that rad3 is the homologue of Saccharomyces cerevisiae ESR1 (MEC1/SAD3) and Drosophila melanogaster mei-41 checkpoint genes. This establishes Rad3/Mec1 as the only conserved protein which is required for all the DNA structure checkpoints in both yeast model systems. Rad3 is an inessential member of the 'lipid kinase' subclass of kinases which includes the ATM protein defective in ataxia telangiectasia patients. Mutational analysis indicates that the kinase domain is required for Rad3 function, and immunoprecipitation of overexpressed Rad3 demonstrates an associated protein kinase activity. The previous observation that rad3 mutations can be rescued by a truncated clone lacking the kinase domain may be due to intragenic complementation. Consistent with this, biochemical data suggest that Rad3 exists in a complex containing multiple copies of Rad3. We have identified a novel human gene (ATR) whose product is closely related to Rad3/Esr1p/Mei-41. ATR can functionally complement esr1-1 radiation sensitivity in S. cerevisiae. Together, the structural conservation and functional complementation suggest strongly that the mechanisms underlying the DNA structure checkpoints are conserved throughout evolution.

  • versatile shuttle vectors and genomic libraries for use with Schizosaccharomyces Pombe
    Gene, 1992
    Co-Authors: N C Barbet, W J Muriel, Antony M Carr
    Abstract:

    We have constructed a variety of pUC-based vectors designed for maintenance in Schizosaccharomyces Pombe. These can be used for both gene bank construction and subcloning. Plasmids pUR18 and pUR19 are modifications of pUC vectors containing the Sc. Pombe ars1 and ura4 sequences and retaining the lacZ XGal blue-white selection system for screening for DNA inserts. These vectors have been used to construct representative Sc. Pombe and Saccharomyces cerevisiae genomic libraries. To assist in the creation of gene deletions, we have constructed another two plasmids. Combined with the technique of partially filling-in 5' overhangs created with restriction enzymes, these plasmids simplify the replacement of all or part of an open reading frame by a functional ura4 gene. Furthermore, such constructs can be excised with SfiI as a linear fragment for use in Sc. Pombe transformations. When integrated into the Sc. Pombe genome, the site of integration can be easily mapped by pulsed-field gel electrophoresis using the presence of a novel NotI site.

  • dna repair mutants defining g2 checkpoint pathways in Schizosaccharomyces Pombe
    The EMBO Journal, 1992
    Co-Authors: F Alkhodairy, Antony M Carr
    Abstract:

    We have tested mutants corresponding to 20 DNA repair genes of the fission yeast Schizosaccharomyces Pombe for their ability to arrest in G2 after DNA damage. Of the mutants tested, four are profoundly defective in this damage dependent G2 arrest. In addition, these four mutants are highly sensitive to a transient inhibition of DNA synthesis by hydroxyurea. This suggests that the pathway responsible for the recognition of DNA damage and the subsequent mitotic arrest, shares many functions with the mechanism that controls the dependency of mitosis on the completion of S phase. The phenotype of these checkpoint rad mutants in wee mutant backgrounds indicate that the G2 arrest response is mediated either through, or in parallel with, the activity of the cdc2 gene product.

Edina Csaszar - One of the best experts on this subject based on the ideXlab platform.

  • an improved strategy for tandem affinity purification tagging of Schizosaccharomyces Pombe genes
    Proteomics, 2009
    Co-Authors: Lubos Cipak, Cornelia Rumpf, Karl Mechtler, Mario Spirek, Maria Novatchkova, Zhiming Chen, Wolfgang Lugmayr, Gustav Ammerer, Edina Csaszar
    Abstract:

    Tandem affinity purification (TAP) is a method that allows rapid purification of native protein complexes. We developed an improved technique to fuse the fission yeast genes with a TAP tag. Our technique is based on tagging constructs that contain regions homologous to the target gene cloned into vectors carrying a TAP tag. We used this technique to design strategies for TAP-tagging of predicted Schizosaccharomyces Pombe genes (http://mendel.imp.ac.at/Pombe_tagging/). To validate the approach, we purified the proteins, which associated with two evolutionarily conserved proteins Swi5 and Sfr1 as well as three protein kinases Ksg1, Orb6 and Sid1.

Akihisa Matsuyama - One of the best experts on this subject based on the ideXlab platform.

Related terms

Schizosaccharomyces Pombe - 15 days free trial to Access Experts & Abstracts