scholarly journals Budding yeast centromeric DNA and A+T rich bacterial DNA can function as centromeres in the fission yeast Schizosaccharomyces pombe

2019 ◽  
Author(s):  
Anne C Barbosa ◽  
Zhengyao Xu ◽  
Kazhal Karari ◽  
Silke Hauf ◽  
William RA Brown
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Dna Sequence ◽  
Budding Yeast ◽  
Bacterial Dna ◽  
Centromeric Dna ◽  
Serine Recombinases ◽  
Centromere Function ◽  
And Function

Eukaryotic centromeric DNA is famously variable in evolution but currently, this cannot be reconciled with the conservation of eukaryotic centromere function. It seems likely that centromeric DNA from different organisms contains conserved functionally important features but the identity of these features is unknown. The point centromeres of the budding yeast Saccharomyces cerevisiae and the regional centromeres of the fission yeast Schizosaccharomyces pombe are separated by 350 million years of evolution and are canonical examples of the paradoxical relationship1 between centromeric DNA sequence and function. We have established a centromere-replacement strategy in Schizosaccharomyces pombe in order to resolve this paradox experimentally. Centromere-replacement shows that an A+T rich bacterial DNA sequence has weak centromere function and that elements of the Saccharomyces cerevisiae centromere embedded in short sequences from the non-centromeric S. pombe wee1 gene function almost as well as native S. pombe centromeric DNA. These observations demonstrate that determinants of centromere function are held in common by the budding and fission yeasts and that A+T rich DNA is both necessary and sufficient for function in S. pombe. Given the evolutionary distance between these yeasts, it is likely that A+T rich DNA has centromere function in a wide variety of eukaryotes. Centromere-replacement uses unidirectional serine recombinases that work well in many organisms2 3 and our experimental strategy should allow this idea to be tested in other eukaryotes.

scholarly journals CENP-A exceeds microtubule attachment sites in centromere clusters of both budding and fission yeast

2011 ◽  
Vol 195 (4) ◽  
pp. 563-572 ◽  
Author(s):  
Valerie C. Coffman ◽  
Pengcheng Wu ◽  
Mark R. Parthun ◽  
Jian-Qiu Wu
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Budding Yeast ◽  
Histone H3 ◽  
Kinetochore Assembly ◽  
Microtubule Attachment ◽  
Attachment Sites ◽  
Histone H3 Variant ◽  
Architectural Models

The stoichiometries of kinetochores and their constituent proteins in yeast and vertebrate cells were determined using the histone H3 variant CENP-A, known as Cse4 in budding yeast, as a counting standard. One Cse4-containing nucleosome exists in the centromere (CEN) of each chromosome, so it has been assumed that each anaphase CEN/kinetochore cluster contains 32 Cse4 molecules. We report that anaphase CEN clusters instead contained approximately fourfold more Cse4 in Saccharomyces cerevisiae and ∼40-fold more CENP-A (Cnp1) in Schizosaccharomyces pombe than predicted. These results suggest that the number of CENP-A molecules exceeds the number of kinetochore-microtubule (MT) attachment sites on each chromosome and that CENP-A is not the sole determinant of kinetochore assembly sites in either yeast. In addition, we show that fission yeast has enough Dam1–DASH complex for ring formation around attached MTs. The results of this study suggest the need for significant revision of existing CEN/kinetochore architectural models.

Structural organization and functional analysis of centromeric DNA in the fission yeast Schizosaccharomyces pombe

1988 ◽  
Vol 8 (2) ◽  
pp. 754-763
Author(s):  
B Fishel ◽  
H Amstutz ◽  
M Baum ◽  
J Carbon ◽  
L Clarke
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Dna Sequences ◽  
Centromeric Dna ◽  
Centromere Function ◽  
Leu2 Gene ◽  
Centromeric Repeat ◽  
Sequence Elements ◽  
Chromosome Iii ◽  
Chromosome Ii

Centromeric DNA in the fission yeast Schizosaccharomyces pombe was isolated by chromosome walking and by field inversion gel electrophoretic fractionation of large genomic DNA restriction fragments. The centromere regions of the three chromosomes were contained on three SalI fragments (120 kilobases [kb], chromosome III; 90 kb, chromosome II; and 50 kb, chromosome I). Each fragment contained several repetitive DNA sequences, including repeat K (6.4 kb), repeat L (6.0 kb), and repeat B, that occurred only in the three centromere regions. On chromosome II, these repeats were organized into a 35-kb inverted repeat that included one copy of K and L in each arm of the repeat. Site-directed integration of a plasmid containing the yeast LEU2 gene into K repeats at each of the centromeres or integration of an intact K repeat into a chromosome arm had no effect on mitotic or meiotic centromere function. The centromeric repeat sequences were not transcribed and possessed many of the properties of constitutive heterochromatin. Thus, S. pombe is an excellent model system for studies on the role of repetitive sequence elements in centromere function.

scholarly journals Structural organization and functional analysis of centromeric DNA in the fission yeast Schizosaccharomyces pombe.

1988 ◽  
Vol 8 (2) ◽  
pp. 754-763 ◽  
Author(s):  
B Fishel ◽  
H Amstutz ◽  
M Baum ◽  
J Carbon ◽  
L Clarke
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Dna Sequences ◽  
Centromeric Dna ◽  
Centromere Function ◽  
Leu2 Gene ◽  
Centromeric Repeat ◽  
Sequence Elements ◽  
Chromosome Iii ◽  
Chromosome Ii

Centromeric DNA in the fission yeast Schizosaccharomyces pombe was isolated by chromosome walking and by field inversion gel electrophoretic fractionation of large genomic DNA restriction fragments. The centromere regions of the three chromosomes were contained on three SalI fragments (120 kilobases [kb], chromosome III; 90 kb, chromosome II; and 50 kb, chromosome I). Each fragment contained several repetitive DNA sequences, including repeat K (6.4 kb), repeat L (6.0 kb), and repeat B, that occurred only in the three centromere regions. On chromosome II, these repeats were organized into a 35-kb inverted repeat that included one copy of K and L in each arm of the repeat. Site-directed integration of a plasmid containing the yeast LEU2 gene into K repeats at each of the centromeres or integration of an intact K repeat into a chromosome arm had no effect on mitotic or meiotic centromere function. The centromeric repeat sequences were not transcribed and possessed many of the properties of constitutive heterochromatin. Thus, S. pombe is an excellent model system for studies on the role of repetitive sequence elements in centromere function.

scholarly journals Seg1 controls eisosome assembly and shape

2012 ◽  
Vol 198 (3) ◽  
pp. 405-420 ◽  
Author(s):  
Karen E. Moreira ◽  
Sebastian Schuck ◽  
Bianca Schrul ◽  
Florian Fröhlich ◽  
James B. Moseley ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Plasma Membrane ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Budding Yeast ◽  
Yeast Saccharomyces Cerevisiae ◽  
Normal Plasma ◽  
Cytoplasmic Proteins ◽  
Stable Domains

Eisosomes are stable domains at the plasma membrane of the budding yeast Saccharomyces cerevisiae and have been proposed to function in endocytosis. Eisosomes are composed of two main cytoplasmic proteins, Pil1 and Lsp1, that form a scaffold around furrow-like plasma membrane invaginations. We show here that the poorly characterized eisosome protein Seg1/Ymr086w is important for eisosome biogenesis and architecture. Seg1 was required for efficient incorporation of Pil1 into eisosomes and the generation of normal plasma membrane furrows. Seg1 preceded Pil1 during eisosome formation and established a platform for the assembly of other eisosome components. This platform was further shaped and stabilized upon the arrival of Pil1 and Lsp1. Moreover, Seg1 abundance controlled the shape of eisosomes by determining their length. Similarly, the Schizosaccharomyces pombe Seg1-like protein Sle1 was necessary to generate the filamentous eisosomes present in fission yeast. The function of Seg1 in the stepwise biogenesis of eisosomes reveals striking architectural similarities between eisosomes in yeast and caveolae in mammals.

scholarly journals The Schizosaccharomyces pombe cho1+ Gene Encodes a Phospholipid Methyltransferase

Genetics ◽  
1998 ◽  
Vol 150 (2) ◽  
pp. 553-562
Author(s):  
Margaret I Kanipes ◽  
John E Hill ◽  
Susan A Henry
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Sequence Analysis ◽  
Schizosaccharomyces Pombe ◽  
Gene Disruption ◽  
Structure And Function ◽  
Biosynthetic Enzyme ◽  
Gene Encoding ◽  
Complementation Groups ◽  
Eukaryotic Organisms ◽  
And Function

Abstract The isolation of mutants of Schizosaccharomyces pombe defective in the synthesis of phosphatidylcholine via the methylation of phosphatidylethanolamine is reported. These mutants are choline auxotrophs and fall into two unlinked complementation groups, cho1 and cho2. We also report the analysis of the cho1+ gene, the first structural gene encoding a phospholipid biosynthetic enzyme from S. pombe to be cloned and characterized. The cho1+ gene disruption mutant (cho1Δ) is viable if choline is supplied and resembles the cho1 mutants isolated after mutagenesis. Sequence analysis of the cho1+ gene indicates that it encodes a protein closely related to phospholipid methyltransferases from Saccharomyces cerevisiae and rat. Phospholipid methyltransferases encoded by a rat liver cDNA and the S. cerevisiae OPI3 gene are both able to complement the choline auxotrophy of the S. pombe cho1 mutants. These results suggest that both the structure and function of the phospholipid N-methyltransferases are broadly conserved among eukaryotic organisms.

scholarly journals Upstream – News in Genomics

2002 ◽  
Vol 3 (3) ◽  
pp. 221-225
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ovarian Cancer ◽  
Oryza Sativa ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Proteomic Analysis ◽  
Large Scale ◽  
Protein Complexes ◽  
The Other ◽  
Blood Samples

In recent months a bumper crop of genomes has been completed, including the fission yeast (Schizosaccharomyces pombe) and rice (Oryza sativa). Two large-scale studies ofSaccharomyces cerevisiaeprotein complexes provided a picture of the eukaryotic proteome as a network of complexes. Amongst the other stories of interest was a demonstration that proteomic analysis of blood samples can be used to detect ovarian cancer, perhaps even as early as stage I.

scholarly journals Analysis of centromeric DNA in the fission yeast Schizosaccharomyces pombe.

1986 ◽  
Vol 83 (21) ◽  
pp. 8253-8257 ◽  
Author(s):  
L. Clarke ◽  
H. Amstutz ◽  
B. Fishel ◽  
J. Carbon
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Centromeric Dna
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