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A 4906-bp DNA fragment, which complemented Schizosaccharomyces pombe strains partly defective in pyruvate dehydrogenase (PDH), was cloned and sequenced. The fragment contained an open reading frame (ORF) of 366 amino acids (aa), which showed 62% identity with the Saccharomyces cerevisiae gene encoding the PDH E1β subunit, and significant similarity to subunits from a number of other 2-oxo-acid dehydrogenase...
Here we report the isolation of an enolase (Eno)-encoding cDNA clone from Schizosaccharomyces pombe. The deduced amino acid (aa) sequence of the 1.4-kb cDNA shares identities with a number of Eno from Escherichia coli to humans. The highest degree of similarity is to the known Eno from Saccharomyces cerevisiae and an Eno from Candida albicans. Northern blot analysis identified a single transcript...
We have constructed a new multipurpose stable shuttle vector for the fission yeast Schizosaccharomyces pombe (Sp). Plasmid pDblet was designed to provide convenient features for molecular work and to overcome the inconveniences of previously designed Sp vectors. It contains the Sp ura4 gene as selectable marker and a new highly efficient ARS (autonomously replicating sequence) element, allowing...
Mitochondrial (mt) DNA replication is carried out by the nuclear-encoded DNA polymerase-γ (Pol-γ). We have cloned a new DNA polymerase-encoding gene from Schizosaccharomyces pombe (Sp), which we believe encodes the homologue of the Saccharomyces cerevisiae (Sc) mt DNA polymerase (MIP1). The putative Sp polγ gene expressed a transcript of approx. 4-kb that contained a 3-kb open reading frame encoding...
Major transitions in the eukaryotic cell cycle are regulated by the cyclin-dependent protein kinases (CDK). In particular, the G2/M transition is initiated by the activity of a complex formed by a CDK of the Cdc2/Cdc28 family and B-type cyclins of the Cdc13/Clb family in the yeasts, Schizosaccharomyces pombe (Sp) and Saccharomyces cerevisiae (Sc). To study the molecular mechanisms that control the...
Defects in the Schizosaccharomyces pombe (Sp) cell cycle-controlling genes prevent the cell cycle progression. Mutations in one of the late septation genes, cdc4, cause Sp cells to arrest at cytokinesis and result in an elongated cellular morphology. By functional complementation of one of the temperature-sensitive (ts) mutant alleles of cdc4, cdc4-31, with a Sp cDNA library, a novel gene, scf1,...
The construction of vectors for use in Schizosaccharomyces pombe using the his3 + gene as a selectable marker is described. In addition, we report the construction of a genomic library in a his3 + -containing shuttle vector to facilitate the cloning of genes by complementation of mutant function in strains defective for His3 activity.
In the course of the Schizosaccharomyces pombe cDNA project, we succeeded in cloning all the genes encoding translation elongation factors EF-1α, EF-1β, EF-1γ, EF-2 and EF-3. With the exception of the EF-1γ gene, the nucleotide (nt) sequence ofS. pombe elongation factors has not been previously reported. For EF-1α, we found three genes whose amino acid (aa) sequences are quite homologous each other...
RNA polymerase II purified from the fission yeast Schizosaccharomyces pombe contains 10 different species of polypeptides. Previously, we cloned and sequenced both cDNA and the genes encoding the four large subunits, Rpb1, Rpb2, Rpb3 and Rpb5. Later, other groups isolated the genes for Rpb6 and Rpb12 and cDNA for Rpb10. Here, we cloned both cDNA and the genes encoding four small subunits, Rpb7, Rpb8,...
A Schizosaccharomyces pombe temperature-sensitive mutant, rae1-1, was previously identified by us as being defective in nuclear export of Poly(A) + RNA when grown at restrictive temperature. Here, we report the isolation of the human homologue of the S. pombe rae1 gene. The RAE1 genes are highly conserved in evolution in both structure and function. The human RAE1 cDNA, when expressed from...
We report on the cloning of sep1 + , a gene whose mutation causes filamentous growth in Schizosaccharomyces pombe. Since cell growth and propagation are not affected by the mutation, it could not be cloned using selective conditions for the identification of the positive transformants. Instead, we cloned it from a cosmid of a contig (Hoheisel et al., Cell 73, 109-1120, 1993) supposed to cover...
A novel heat shock protein 70 (HSP70) gene, pss1 + , of fission yeast, Schizosaccharomyces pombe (S. pombe), has been isolated as a multicopy suppressor of a synthetic lethal mutant of ras1 + , which shows severe retardation of growth and aggregation phenotype when the ras1 gene function is absent. The pss1 + gene functionally complements the growth defect of the mutant. Sequence...
In contrast to homologous genes in other fungal mitochondrial genomes, the gene encoding subunit 2 of cytochrome oxidase (cox2) in several Schizosaccharomyces pombe strains contains a large group II intron. Its 2436 nucleotides can be folded into a typical group II intron secondary structure, possessing all the expected sequence motifs for subgroup IIA1 (Michel et al., 1989). This intron is remarkable...
Both the rpb9 gene and its cDNA encoding the subunit 9 of RNA polymerase II were cloned from the fission yeast Schizosaccharomyces pombe. From the DNA sequences, Rpb9 was predicted to consist of 113 amino acid residues with a molecular mass of 13 175. S. pombe Rpb9 is 47, 40 and 36% identical in amino acid sequence to the corresponding subunits from Saccharomyces cerevisiae, human and Drosophila...
A series of vectors is described which enables the episomal expression of proteins fused to different tag sequences in Schizosaccharomyces pombe. Proteins can be expressed with their amino termini fused to GFP/EGFP, three copies of the HA or Pk epitopes or a combined tag which contains two copies of the myc epitope and six histidine residues (MH). Fusion of the carboxyl terminus of a protein to a...
Reversible tyrosine phosphorylation of proteins is one of the major regulatory physiological events in response to cell-cell- and cell-matrix contact in Metazoa. Previously it was documented that the tyrosine phosphorylating enzymes, the tyrosine kinases (TKs), are autapomorphic characters of Metazoa, including sponges. In this paper the tyrosine dephosphorylating enzymes, the protein-tyrosine phosphatases...
The increasing use of the fission yeast Schizosaccharomyces pombe as a model organism for elucidating the mechanisms of critical biological processes such as cell-cycle control, DNA replication, and stress-mediated signal transduction has fostered the development and utilization of expression systems for gene function analysis. Using the promoter of the ctr4 + copper transporter gene from...
The fork-head type transcription factors are a class of regulators that function in a broad spectrum of cellular and developmental processes in many species ranging from yeasts to human. Previous data on yeast fork-head genes suggested roles for these regulators in the control of cell division, sexual differentiation and development. The genome of Schizosaccharomyces pombe has four genes that code...
The Rec12 (Spo11) protein of the fission yeast Schizosaccharomyces pombe is a meiosis-specific ortholog of the catalytic subunit of type VI topoisomerases and is thought to catalyze double-strand DNA breaks that initiate recombination. We tested the hypothesis that the rec12-117 allele affects the choice of pathways by which recombination is resolved. DNA sequence analysis revealed a single missense...
Integration of retroviral elements into the host genome is a phenomena observed among many classes of retroviruses. Much information concerning the integration of retroviral elements has been documented based on in vitro analysis or expression of selectable markers. To identify possible Tf1 integration events within silent regions of the Schizosaccharomyces pombe genome, we focused on performing an...
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