scholarly journals The endo-beta-1,3-glucanase eng1p is required for dissolution of the primary septum during cell separation in Schizosaccharomyces pombe

2003 ◽  
Vol 116 (9) ◽  
pp. 1689-1698 ◽  
Author(s):  
A. B. Martin-Cuadrado
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Cell Separation ◽  
Primary Septum

scholarly journals Scw1p Antagonizes the Septation Initiation Network To Regulate Septum Formation and Cell Separation in the Fission Yeast Schizosaccharomyces pombe

2003 ◽  
Vol 2 (3) ◽  
pp. 510-520 ◽  
Author(s):  
Quan-Wen Jin ◽  
Dannel McCollum
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Cell Separation ◽  
Deletion Mutant ◽  
Temperature Sensitive ◽  
Growth Defects ◽  
Septum Formation ◽  
Primary Septum ◽  
Septation Initiation Network ◽  
Ring Constriction

ABSTRACT Cytokinesis in the fission yeast Schizosaccharomyces pombe is regulated by a signaling pathway termed the septation initiation network (SIN). The SIN is essential for initiation of actomyosin ring constriction and septum formation. In a screen to search for mutations that can rescue the sid2-250 SIN mutant, we obtained scw1-18. Both the scw1-18 mutant and the scw1 deletion mutant (scw1Δ mutant), have defects in cell separation. Both the scw1-18 and scw1Δ mutations rescue the growth defects of not just the sid2-250 mutant but also the other temperature-sensitive SIN mutants. Other cytokinesis mutants, such as those defective for actomyosin ring formation, are not rescued by scw1Δ. scw1Δ does not seem to rescue the SIN by restoring SIN signaling defects. However, scw1Δ may function downstream of the SIN to promote septum formation, since scw1Δ can rescue the septum formation defects of the cps1-191β-1,3-glucan synthase mutant, which is required for synthesis of the primary septum.

scholarly journals Rho3p Regulates Cell Separation by Modulating Exocyst Function in Schizosaccharomyces pombe

Genetics ◽  
2003 ◽  
Vol 164 (4) ◽  
pp. 1323-1331
Author(s):  
Hongyan Wang ◽  
Xie Tang ◽  
Mohan K Balasubramanian
Keyword(s):  
Cell Division ◽  
Schizosaccharomyces Pombe ◽  
Cell Separation ◽  
Model Organism ◽  
Temperature Sensitive ◽  
Complex Cells ◽  
Exocyst Complex ◽  
Primary Septum ◽  
Rho Family ◽  
Growth Phenotype

Abstract Cytokinesis is the final stage of the cell division cycle in which the mother cell is physically divided into two daughters. In recent years the fission yeast Schizosaccharomyces pombe has emerged as an attractive model organism for the study of cytokinesis, since it divides using an actomyosin ring whose constriction is coordinated with the centripetal deposition of new membranes and a division septum. The final step of cytokinesis in S. pombe requires the digestion of the primary septum to liberate two daughters. We have previously shown that the multiprotein exocyst complex is essential for this process. Here we report the isolation of rho3+, encoding a Rho family GTPase, as a high-copy suppressor of an exocyst mutant, sec8-1. Overproduction of Rho3p also suppressed the temperature-sensitive growth phenotype observed in cells lacking Exo70p, another conserved component of the S. pombe exocyst complex. Cells deleted for rho3 arrest at higher growth temperatures with two or more nuclei and uncleaved division septa between pairs of nuclei. rho3Δ cells accumulate ∼100-nm vesicle-like structures. These phenotypes are all similar to those observed in exocyst component mutants, consistent with a role for Rho3p in modulation of exocyst function. Taken together, our results suggest the possibility that S. pombe Rho3p regulates cell separation by modulation of exocyst function.

scholarly journals The α-glucanase Agn1p is required for cell separation in Schizosaccharomyces pombe

2005 ◽  
Vol 97 (7) ◽  
pp. 569-576 ◽  
Author(s):  
Ignacio García ◽  
David Jiménez ◽  
Victoria Martín ◽  
Angel Durán ◽  
Yolanda Sánchez
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Cell Separation

Regulation of the start of DNA replication in Schizosaccharomyces pombe

1999 ◽  
Vol 112 (6) ◽  
pp. 939-946 ◽  
Author(s):  
C.R. Carlson ◽  
B. Grallert ◽  
T. Stokke ◽  
E. Boye
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Growth Rate ◽  
Schizosaccharomyces Pombe ◽  
Growth Rates ◽  
Cell Separation ◽  
Cell Mass ◽  
Nitrogen Sources ◽  
S Phase ◽  
G1 Phase

Cells of Schizosaccharomyces pombe were grown in minimal medium with different nitrogen sources under steady-state conditions, with doubling times ranging from 2.5 to 14 hours. Flow cytometry and fluorescence microscopy confirmed earlier findings that at rapid growth rates, the G1 phase was short and cell separation occurred at the end of S phase. For some nitrogen sources, the growth rate was greatly decreased, the G1 phase occupied 30–50% of the cell cycle, and cell separation occurred in early G1. In contrast, other nitrogen sources supported low growth rates without any significant increase in G1 duration. The method described allows manipulation of the length of G1 and the relative cell cycle position of S phase in wild-type cells. Cell mass was measured by flow cytometry as scattered light and as protein-associated fluorescence. The extensions of G1 were not related to cell mass at entry into S phase. Our data do not support the hypothesis that the cells must reach a certain fixed, critical mass before entry into S. We suggest that cell mass at the G1/S transition point is variable and determined by a set of molecular parameters. In the present experiments, these parameters were influenced by the different nitrogen sources in a way that was independent of the actual growth rate.

scholarly journals Phylogenetic and comparative functional analysis of the cell-separation α-glucanase Agn1p in Schizosaccharomyces

Microbiology ◽  
2014 ◽  
Vol 160 (6) ◽  
pp. 1063-1074 ◽  
Author(s):  
Matthias Sipiczki ◽  
Anita Balazs ◽  
Aniko Monus ◽  
Laszlo Papp ◽  
Anna Horvath ◽  
...  
Keyword(s):  
Cell Wall ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Mother Cell ◽  
Cell Separation ◽  
Yeast Cells ◽  
Glycoside Hydrolase Family ◽  
Binding Domains ◽  
Yeast Phase ◽  
Mother Cell Wall

The post-cytokinetic separation of cells in cell-walled organisms involves enzymic processes that degrade a specific layer of the division septum and the region of the mother cell wall that edges the septum. In the fission yeast Schizosaccharomyces pombe, the 1,3-α-glucanase Agn1p, originally identified as a mutanase-like glycoside hydrolase family 71 (GH71) enzyme, dissolves the mother cell wall around the septum edge. Our search in the genomes of completely sequenced fungi identified GH71 hydrolases in Basidiomycota, Taphrinomycotina and Pezizomycotina, but not in Saccharomycotina. The most likely Agn1p orthologues in Pezizomycotina species are not mutanases having mutanase-binding domains, but experimentally non-characterized hypothetical proteins that have no carbohydrate-binding domains. The analysis of the GH71 domains corroborated the phylogenetic relationships of the Schizosaccharomyces species determined by previous studies, but suggested a closer relationship to the Basidiomycota proteins than to the Ascomycota proteins. In the Schizosaccharomyces genus, the Agn1p proteins are structurally conserved: their GH71 domains are flanked by N-terminal secretion signals and C-terminal sequences containing the conserved block YNFNAY/HTG. The inactivation of the agn1Sj gene in Schizosaccharomyces japonicus, the only true dimorphic member of the genus, caused a severe cell-separation defect in its yeast phase, but had no effect on the hyphal growth and yeast-to-mycelium transition. It did not affect the mycelium-to-yeast transition either, only delaying the separation of the yeast cells arising from the fragmenting hyphae. The heterologous expression of agn1Sj partially rescued the separation defect of the agn1Δ cells of Schizosaccharomyces pombe. The results presented indicate that the fission yeast Agn1p 1,3-α-glucanases of Schizosaccharomyces japonicus and Schizosaccharomyces pombe share conserved functions in the yeast phase.

scholarly journals Roles of Hof1p, Bni1p, Bnr1p, and Myo1p in Cytokinesis inSaccharomyces cerevisiae

2000 ◽  
Vol 11 (2) ◽  
pp. 593-611 ◽  
Author(s):  
Elizabeth A. Vallen ◽  
Juliane Caviston ◽  
Erfei Bi
Keyword(s):  
Cell Separation ◽  
Synthetic Lethality ◽  
Temperature Sensitive ◽  
Concerted Action ◽  
Septum Formation ◽  
Primary Septum ◽  
Bud Neck ◽  
Single Ring ◽  
Functional Pathway ◽  
Actomyosin Contraction

Cytokinesis in Saccharomyces cerevisiae occurs by the concerted action of the actomyosin system and septum formation. Here we report on the roles of HOF1,BNI1, and BNR1 in cytokinesis, focusing on Hof1p. Deletion of HOF1 causes a temperature-sensitive defect in septum formation. A Hof1p ring forms on the mother side of the bud neck in G2/M, followed by the formation of a daughter-side ring. Around telophase, Hof1p is phosphorylated and the double rings merge into a single ring that contracts slightly and may colocalize with the actomyosin structure. Upon septum formation, Hof1p splits into two rings, disappearing upon cell separation. Hof1p localization is dependent on septins but not Myo1p. Synthetic lethality suggests that Bni1p and Myo1p belong to one functional pathway, whereas Hof1p and Bnr1p belong to another. These results suggest that Hof1p may function as an adapter linking the primary septum synthesis machinery to the actomyosin system. The formation of the actomyosin ring is not affected by bni1Δ, hof1Δ, orbnr1Δ. However, Myo1p contraction is affected bybni1Δ but not by hof1Δ orbnr1Δ. In bni1Δ cells that lack the actomyosin contraction, septum formation is often slow and asymmetric, suggesting that actomyosin contraction may provide directionality for efficient septum formation.

scholarly journals Role of the α-Glucanase Agn1p in Fission-Yeast Cell Separation

2004 ◽  
Vol 15 (8) ◽  
pp. 3903-3914 ◽  
Author(s):  
Nick Dekker ◽  
Dave Speijer ◽  
Christian H. Grün ◽  
Marlene van den Berg ◽  
Annett de Haan ◽  
...  
Keyword(s):  
Cell Division ◽  
Fission Yeast ◽  
Cell Separation ◽  
Wall Material ◽  
Dependent Manner ◽  
Cellular Functions ◽  
Daughter Cells ◽  
Primary Septum ◽  
Cycle Dependent ◽  
Division Cell

Cell division in the fission yeast Schizosaccharomyces pombe yields two equal-sized daughter cells. Medial fission is achieved by deposition of a primary septum flanked by two secondary septa within the dividing cell. During the final step of cell division, cell separation, the primary septum is hydrolyzed by an endo-(1,3)-β-glucanase, Eng1p. We reasoned that the cell wall material surrounding the septum, referred to here as the septum edging, also must be hydrolyzed before full separation of the daughter cells can occur. Because the septum edging contains (1,3)-α-glucan, we investigated the cellular functions of the putative (1,3)-α-glucanases Agn1p and Agn2p. Whereas agn2 deletion results in a defect in endolysis of the ascus wall, deletion of agn1 leads to clumped cells that remained attached to each other by septum-edging material. Purified Agn1p hydrolyzes (1,3)-α-glucan predominantly into pentasaccharides, indicating an endo-catalytic mode of hydrolysis. Furthermore, we show that the transcription factors Sep1p and Ace2p regulate both eng1 and agn1 expression in a cell cycle-dependent manner. We propose that Agn1p acts in concert with Eng1p to achieve efficient cell separation, thereby exposing the secondary septa as the new ends of the daughter cells.

scholarly journals Cdc42 promotes Bgs1 recruitment for septum synthesis and glucanase localization for cell separation during cytokinesis in fission yeast

2019 ◽  
Author(s):  
Udo N. Onwubiko ◽  
Julie Robinson ◽  
Rose Albu Mustaf ◽  
Maitreyi E. Das
Keyword(s):  
Fission Yeast ◽  
Membrane Trafficking ◽  
Cell Separation ◽  
Nucleotide Exchange ◽  
Timely Initiation ◽  
Division Site ◽  
Guanine Nucleotide Exchange ◽  
Nucleotide Exchange Factors ◽  
Primary Septum ◽  
Ring Constriction

AbstractCytokinesis in fission yeast involves actomyosin ring constriction concurrent to septum synthesis followed by septum digestion resulting in cell separation. A recent report indicates that endocytosis is required for septum synthesis and cell separation. The conserved GTPase Cdc42 is required for membrane trafficking and promotes endocytosis. Cdc42 is activated by Guanine nucleotide exchange factors (GEFs). Cdc42 GEFs have been shown to promote timely initiation of septum synthesis and proper septum morphology. Here we show that Cdc42 promotes the recruitment of the major primary septum synthesizing enzyme Bgs1 and consequent ring constriction. Cdc42 is also required for proper localization of the septum digesting glucanases at the division site. Thus, Cdc42 is required to promote multiple steps during cytokinesis.

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