scholarly journals Fission yeast TORC1 prevents eIF2  phosphorylation in response to nitrogen and amino acids via Gcn2 kinase

2012 ◽  
Vol 125 (24) ◽  
pp. 5955-5959 ◽  
Author(s):  
N. Valbuena ◽  
A. E. Rozalen ◽  
S. Moreno
Keyword(s):  
Amino Acids ◽  
Fission Yeast ◽  
Eif2 Phosphorylation

scholarly journals Amino acids whose intracellular levels change most during aging alter chronological lifespan of fission yeast

2020 ◽  
Author(s):  
Charalampos Rallis ◽  
Michael Mülleder ◽  
Graeme Smith ◽  
Yan Zi Au ◽  
Markus Ralser ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Fission Yeast ◽  
Yeast Cells ◽  
Total Amino Acid ◽  
Wild Type ◽  
Chronological Lifespan ◽  
Biomarkers Of Aging ◽  
Concentration Changes ◽  
Amino Acid Levels

AbstractAmino acid deprivation or supplementation can affect cellular and organismal lifespan, but we know little about the role of concentration changes in free, intracellular amino acids during aging. Here, we determine free amino-acid levels during chronological aging of non-dividing fission yeast cells. We compare wild-type with long-lived mutant cells that lack the Pka1 protein of the protein kinase A signalling pathway. In wild-type cells, total amino-acid levels decrease during aging, but much less so in pka1 mutants. Two amino acids strongly change as a function of age: glutamine decreases, especially in wild-type cells, while aspartate increases, especially in pka1 mutants. Supplementation of glutamine is sufficient to extend the chronological lifespan of wild-type but not of pka1Δ cells. Supplementation of aspartate, on the other hand, shortens the lifespan of pka1Δ but not of wild-type cells. Our results raise the possibility that certain amino acids are biomarkers of aging, and their concentrations during aging can promote or limit cellular lifespan.

scholarly journals Phosphorylation of Arp2 is not essential for Arp2/3 complex activity in fission yeast

2018 ◽  
Vol 1 (5) ◽  
pp. e201800202 ◽  
Author(s):  
Alexander E Epstein ◽  
Sofia Espinoza-Sanchez ◽  
Thomas D Pollard
Keyword(s):  
Mass Spectrometry ◽  
Amino Acids ◽  
Fission Yeast ◽  
Aspartic Acid ◽  
Actin Filaments ◽  
Yeast Genome ◽  
Complex Activity ◽  
Actin Patch

LeClaire et al presented evidence that phosphorylation of three sites on the Arp2 subunit activates the Arp2/3 complex to nucleate actin filaments. We mutated the homologous residues of Arp2 (Y198, T233, and T234) in the fission yeast genome to amino acids that preclude or mimic phosphorylation. Arp2/3 complex is essential for the viability of fission yeast, yet strains unable to phosphorylate these sites grew normally. Y198F/T233A/T234A Arp2 was only nonfunctional if GFP-tagged, as observed by LeClaire et al in Drosophila cells. Replacing both T233 and T234 with aspartic acid was lethal, suggesting that phosphorylation might be inhibitory. Nevertheless, blocking phosphorylation at these sites had the same effect as mimicking it: slowing assembly of endocytic actin patches. Mass spectrometry revealed phosphorylation at a fourth conserved Arp2 residue, Y218, but both blocking and mimicking phosphorylation of Y218 only slowed actin patch assembly slightly. Therefore, phosphorylation of Y198, T233, T234, and Y218 is not required for the activity of fission yeast Arp2/3 complex.

Cloning and sequencing of the cyclin-related cdc13+ gene and a cytological study of its role in fission yeast mitosis

1988 ◽  
Vol 91 (4) ◽  
pp. 587-595 ◽  
Author(s):  
I. Hagan ◽  
J. Hayles ◽  
P. Nurse
Keyword(s):  
Amino Acids ◽  
Protein Kinase ◽  
Fission Yeast ◽  
Gene Function ◽  
Mammalian Cells ◽  
Major Part ◽  
Cytological Study ◽  
Partial Loss ◽  
Cloning And Sequencing ◽  
Microtubular Cytoskeleton

We have cloned and sequenced the cdc13+ gene from fission yeast. When a major part of the cdc13+ gene is deleted from the chromosome, cells arrest in interphase, but partial loss of gene activity leads to cells containing condensed chromosomes, aberrant septa and a microtubular cytoskeleton with characteristics of both G2 and M. Expression of this phenotype is influenced by the nutritional status of the cell. Our results suggest that the cdc13+ gene function is required for the control of the G2 to M transition. It appears to play a role in regulating the separate pathways of events involved in the physical process of mitosis, for example in the reorganization of the cytoskeleton on transition from G2 to mitosis. The cdc13+ gene function interacts closely with both the yeast and human homologues of cdc2+, suggesting that mammalian cells may contain a cdc13+ homologue. The gene encodes a putative polypeptide of 482 amino acids, and a central region of 176 amino acids of this polypeptide is 50% identical with sea urchin cyclin. Therefore, the cdc13+ protein is cyclin related and could act as a regulator or substrate of the p34cdc2 protein kinase, which initiates mitosis.

scholarly journals Phosphorylation of Arp2 is not essential for Arp2/3 complex activity in fission yeast

2018 ◽  
Author(s):  
Alexander E. Epstein ◽  
Sofia Espinoza-Sanchez ◽  
Thomas D. Pollard
Keyword(s):  
Mass Spectrometry ◽  
Amino Acids ◽  
Fission Yeast ◽  
Aspartic Acid ◽  
Actin Filaments ◽  
Yeast Genome ◽  
Complex Activity ◽  
Actin Patch

AbstractLeClaire et al. presented evidence that phosphorylation of three sites on the Arp2 subunit activates Arp2/3 complex to nucleate actin filaments. We mutated the homologous residues of Arp2 (Y198, T233 and T234) in the fission yeast genome to amino acids that preclude or mimic phosphorylation. Arp2/3 complex is essential for the viability of fission yeast, yet strains unable to phosphorylate these sites grew normally. Y198F/T233A/T234A Arp2 was only nonfunctional if GFP-tagged, as observed by LeClaire et al. in Drosophila cells. Replacing both T233 and T234 with aspartic acid was lethal, suggesting that phosphorylation might be inhibitory. Nevertheless, blocking phosphorylation at these sites had the same effect as mimicking it: slowing assembly of endocytic actin patches. Mass spectrometry revealed phosphorylation at a fourth conserved Arp2 residue, Y218, but both blocking and mimicking phosphorylation of Y218 only slowed actin patch assembly slightly. Therefore, phosphorylation of Y198, T233, T234 and Y218 is not required for the activity of fission yeast Arp2/3 complex.SummaryPrevious research concluded that phosphorylation at three sites on Arp2 is necessary to activate Arp2/3 complex. Epstein et al. make genomic substitutions blocking or mimicking phosphorylation to demonstrate that phosphorylation of these three sites does not regulate Arp2/3 complex in fission yeast.

scholarly journals Construction, Growth, and Harvesting of Fission Yeast Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) Strains

2017 ◽  
Vol 2017 (6) ◽  
pp. pdb.prot091678 ◽  
Author(s):  
André Koch ◽  
Claudia C. Bicho ◽  
Weronika E. Borek ◽  
Alejandro Carpy ◽  
Boris Maček ◽  
...  
Keyword(s):  
Amino Acids ◽  
Cell Culture ◽  
Stable Isotope ◽  
Fission Yeast ◽  
Isotope Labeling ◽  
Stable Isotope Labeling

scholarly journals A functional 125-kDa core polypeptide of fission yeast DNA topoisomerase II.

1991 ◽  
Vol 11 (12) ◽  
pp. 6093-6102 ◽  
Author(s):  
K Shiozaki ◽  
M Yanagida
Keyword(s):  
Amino Acids ◽  
Fission Yeast ◽  
Topoisomerase Ii ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Limited Proteolysis ◽  
Dna Topoisomerase Ii ◽  
Dna Topoisomerase ◽  
The Core ◽  
Topo Ii

We purified fission yeast DNA topoisomerase II (topo II) to apparent homogeneity. It consists of a single 165-kDa polypeptide in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and, upon treatment with a bifunctional reagent, doubles its molecular weight. Limited proteolysis of intact topo II by papain produces a 125-kDa core, which lacks the N-terminal 75 and the C-terminal approximately 260 amino acids but still contains regions similar to those of bacterial or phage T4 topo II subunits. The core retains relaxing and unknotting activities. Further digestion inactivates the core, cleaving it at the middle of the GyrB-like domain and at the beginning of the GyrA-like domain. Therefore, papain appears to cleave spatially distinct subdomains of topo II. We made top2 mutant genes deleted of the C-terminal 286 or N-terminal 74 amino acids, which can substitute for the wild-type top2+ gene in mitosis and meiosis. However, a mutant containing deletions of both termini cannot rescue the top2 null mutant, despite the fact that the product is enzymatically active. Therefore, the top2 product of the doubly truncated gene may not fulfill all of the in vivo requirements for top2+ function.

scholarly journals Mitochondrial respiration is required to provide amino acids during fermentative proliferation of fission yeast

EMBO Reports ◽  
2020 ◽  
Vol 21 (11) ◽  
Author(s):  
Michal Malecki ◽  
Stephan Kamrad ◽  
Markus Ralser ◽  
Jürg Bähler
Keyword(s):  
Amino Acids ◽  
Fission Yeast ◽  
Mitochondrial Respiration

scholarly journals Amino Acids Whose Intracellular Levels Change Most During Aging Alter Chronological Life Span of Fission Yeast

2020 ◽  
Author(s):  
Charalampos Rallis ◽  
Michael Mülleder ◽  
Graeme Smith ◽  
Yan Zi Au ◽  
Markus Ralser ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Life Span ◽  
Fission Yeast ◽  
Total Amino Acid ◽  
Wild Type ◽  
Biomarkers Of Aging ◽  
Chronological Life Span ◽  
Concentration Changes ◽  
Amino Acid Levels

Abstract Amino acid deprivation or supplementation can affect cellular and organismal life span, but we know little about the role of concentration changes in free, intracellular amino acids during aging. Here, we determine free amino acid levels during chronological aging of nondividing fission yeast cells. We compare wild-type with long-lived mutant cells that lack the Pka1 protein of the protein kinase A signalling pathway. In wild-type cells, total amino acid levels decrease during aging, but much less so in pka1 mutants. Two amino acids strongly change as a function of age: glutamine decreases, especially in wild-type cells, while aspartate increases, especially in pka1 mutants. Supplementation of glutamine is sufficient to extend the chronological life span of wild-type but not of pka1Δ cells. Supplementation of aspartate, on the other hand, shortens the life span of pka1Δ but not of wild-type cells. Our results raise the possibility that certain amino acids are biomarkers of aging, and their concentrations during aging can promote or limit cellular life span.

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