scholarly journals Isolation of temperature-sensitive diphtheria toxins in yeast and their effects on Drosophila cells

Development ◽  
1992 ◽  
Vol 114 (3) ◽  
pp. 787-796 ◽  
Author(s):  
H.J. Bellen ◽  
D. D'Evelyn ◽  
M. Harvey ◽  
S.J. Elledge
Keyword(s):  
Photoreceptor Cells ◽  
Temperature Sensitive ◽  
Cellular Functions ◽  
Yeast Saccharomyces Cerevisiae ◽  
Similar Temperature Dependence ◽  
Wide Range ◽  
Similar Temperature ◽  
Arrest Growth ◽  
Adp Ribosyltransferase ◽  
Drosophila Cells

We have isolated temperature-sensitive diphtheria toxins (DT-A(ts)) to develop a method that allows temporal impedement of cellular functions. Four DT-A(ts) genes were isolated in a mutagenesis screen using the yeast, Saccharomyces cerevisiae. When expressed in yeast, these DT-A(ts) arrest growth at 18 degrees C but not at 30 degrees C. Three DT-A(ts) were subsequently tested in the R1-R6 photoreceptor cells of transgenic fruit flies, Drosophila melanogaster. The toxins show similar temperature dependence in both organisms, suggesting that they may be useful in a wide range of non-homeothermic species. DNA sequence analysis revealed that three of the four DT-A(ts) mutations are novel. Interestingly, the fourth DT-A(ts) carries the same point mutation as the extensively characterized CRM197, an ADP ribosyltransferase-defective form of diphtheria toxin.

scholarly journals Coupling between fast and slow oscillator circuits in Cancer borealis is temperature-compensated

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Daniel Powell ◽  
Sara A Haddad ◽  
Srinivas Gorur-Shandilya ◽  
Eve Marder
Keyword(s):  
Integer Number ◽  
Temperature Sensitive ◽  
Gastric Mill ◽  
Temperature Dependent ◽  
Cancer Borealis ◽  
Cycle Frequency ◽  
Temperature Changes ◽  
Wide Range ◽  
Similar Temperature ◽  
Increasing Temperature

Coupled oscillatory circuits are ubiquitous in nervous systems. Given that most biological processes are temperature-sensitive, it is remarkable that the neuronal circuits of poikilothermic animals can maintain coupling across a wide range of temperatures. Within the stomatogastric ganglion (STG) of the crab, Cancer borealis, the fast pyloric rhythm (~1 Hz) and the slow gastric mill rhythm (~0.1 Hz) are precisely coordinated at ~11°C such that there is an integer number of pyloric cycles per gastric mill cycle (integer coupling). Upon increasing temperature from 7°C to 23°C, both oscillators showed similar temperature-dependent increases in cycle frequency, and integer coupling between the circuits was conserved. Thus, although both rhythms show temperature-dependent changes in rhythm frequency, the processes that couple these circuits maintain their coordination over a wide range of temperatures. Such robustness to temperature changes could be part of a toolbox of processes that enables neural circuits to maintain function despite global perturbations.

scholarly journals Coupling between fast and slow oscillator circuits in Cancer borealis is temperature compensated

2020 ◽  
Author(s):  
D.J. Powell ◽  
S.A. Haddad ◽  
S. Gorur-Shandilya ◽  
E. Marder
Keyword(s):  
Integer Number ◽  
Temperature Sensitive ◽  
Gastric Mill ◽  
Temperature Dependent ◽  
Cancer Borealis ◽  
Cycle Frequency ◽  
Temperature Changes ◽  
Wide Range ◽  
Similar Temperature ◽  
Increasing Temperature

AbstractCoupled oscillatory circuits are ubiquitous in nervous systems. Given that most biological processes are temperature sensitive, it is remarkable that the neuronal circuits of poikilothermic animals can maintain coupling across a wide range of temperatures. Within the stomatogastric ganglion (STG) of the crab, Cancer borealis, the fast pyloric rhythm (~1Hz) and the slow gastric mill rhythm (~0.1Hz) are precisely coordinated at ~11°C such that there is an integer number of pyloric cycles per gastric mill cycle (integer coupling). Upon increasing temperature from 7-23°C, both oscillators showed similar temperature-dependent increases in cycle frequency, and integer coupling between the circuits was conserved. Thus, although both rhythms show temperature dependent changes in rhythm frequency, the processes that couple these circuits maintain their coordination over a wide range of temperature. Such robustness to temperature changes could be part of a toolbox of processes that enables neural circuits to maintain function despite global perturbations.

scholarly journals Sirtuins and their role as physiological modulators of metabolism

2020 ◽  
Vol 74 ◽  
pp. 489-497
Author(s):  
Grażyna Sygitowicz ◽  
Dariusz Sitkiewicz
Keyword(s):  
Energy Homeostasis ◽  
Metabolic Diseases ◽  
Whole Body ◽  
Acid Oxidation ◽  
Special Role ◽  
Cellular Functions ◽  
Current State ◽  
Expression Of Genes ◽  
Wide Range ◽  
Adp Ribosyltransferase

The sirtuins are a family of highly evolutionary conserved NAD+-dependent deacetylases (SIRT1, 2, 3, 5). Certain human sirtuins (SIRT4, 6) have, in addition, an ADP-ribosyltransferase activity. SIRT1 and SIRT2 are located in the nucleus and cytoplasm; SIRT3 exists predominantly in mitochondria, and SIRT6 is located in the nucleus. The mammalian sirtuins have emerged as key metabolic sensors that directly link environmental nutrient signals to metabolic homeostasis. SIRT1 is involved in the regulation of gluconeogenesis and fatty acid oxidation, as well as inhibiting lipogenesis and inflammation in the liver. In addition, they contribute to the mobilization of fat in white adipose tissue, sense nutrient availability in the hypothalamus; regulate insulin secretion in the pancreas; as well as modulating the expression of genes responsible for the activity of the circadian clock in metabolic tissues. Sirtuins are implicated in a variety of cellular functions ranging from gene silencing, through the control of the cell cycle, to energy homeostasis. Caloric restriction, supported by polyphenols, including resveratrol, which is the SIRT1 activator, plays a special role in maintaining energy homeostasis. On a whole body level, the wide range of cellular activities of the sirtuins suggests that they could constitute a therapeutic target to combat obesity and related metabolic diseases. In addition, this work presents the current state of knowledge in the field of sirtuin activity in relation to nutritional status and lifespan.

The kinetically effective stoichiometry of reactions in the carbon-sulphur dioxide system

1973 ◽  
Vol 26 (4) ◽  
pp. 723 ◽  
Author(s):  
JD Blackwood ◽  
DJ McCarthy
Keyword(s):  
Carbon Dioxide ◽  
Sulphur Dioxide ◽  
Elevated Temperatures ◽  
Carbon Disulphide ◽  
Temperature Sensitive ◽  
Similar Temperature Dependence ◽  
Similar Temperature ◽  
Carbon Dioxide Reaction ◽  
Carbonyl Sulphide ◽  
Overall Reactions

The kinetically effective stoichiometry of reactions resulting from contacting carbon and sulphur dioxide at elevated temperatures involves the five overall reactions: 2C + 2SO2 → 2CO2 + S2 C+ S2 + CO2 → 2COS C+ 2COS → CS2 + 2CO C+ COz -, 2C0 C+S, -+ CS, The rates of the two reactions forming carbon disulphide are shown to have similar temperature dependence in the range 800-950�C, while the carbon-carbon dioxide reaction is shown to be more temperature sensitive than the carbon-carbonyl sulphide reaction in the same temperature range. The existence of parallel routes for carbon disulphide formation was detected by using a mechanistic argument to remove the redundancy in the system where carbon disulphide was concerned.

scholarly journals Iron in Translation: From the Beginning to the End

2021 ◽  
Vol 9 (5) ◽  
pp. 1058
Author(s):  
Antonia María Romero ◽  
María Teresa Martínez-Pastor ◽  
Sergi Puig
Keyword(s):  
Translational Regulation ◽  
Translation Termination ◽  
Protein Biosynthesis ◽  
Regulatory Protein ◽  
Dynamic Control ◽  
Essential Element ◽  
Cellular Functions ◽  
Yeast Saccharomyces Cerevisiae ◽  
Eukaryotic Translation ◽  
Responsive Element

Iron is an essential element for all eukaryotes, since it acts as a cofactor for many enzymes involved in basic cellular functions, including translation. While the mammalian iron-regulatory protein/iron-responsive element (IRP/IRE) system arose as one of the first examples of translational regulation in higher eukaryotes, little is known about the contribution of iron itself to the different stages of eukaryotic translation. In the yeast Saccharomyces cerevisiae, iron deficiency provokes a global impairment of translation at the initiation step, which is mediated by the Gcn2-eIF2α pathway, while the post-transcriptional regulator Cth2 specifically represses the translation of a subgroup of iron-related transcripts. In addition, several steps of the translation process depend on iron-containing enzymes, including particular modifications of translation elongation factors and transfer RNAs (tRNAs), and translation termination by the ATP-binding cassette family member Rli1 (ABCE1 in humans) and the prolyl hydroxylase Tpa1. The influence of these modifications and their correlation with codon bias in the dynamic control of protein biosynthesis, mainly in response to stress, is emerging as an interesting focus of research. Taking S. cerevisiae as a model, we hereby discuss the relevance of iron in the control of global and specific translation steps.

scholarly journals A Mutation of the Yeast Gene Encoding PCNA Destabilizes Both Microsatellite and Minisatellite DNA Sequences

Genetics ◽  
1999 ◽  
Vol 151 (2) ◽  
pp. 511-519 ◽  
Author(s):  
Robert J Kokoska ◽  
Lela Stefanovic ◽  
Andrew B Buermeyer ◽  
R Michael Liskay ◽  
Thomas D Petes
Keyword(s):  
Dna Polymerase ◽  
Repetitive Dna ◽  
Dna Sequences ◽  
Repeat Unit ◽  
Nuclear Antigen ◽  
Temperature Sensitive ◽  
Dinucleotide Repeats ◽  
Yeast Saccharomyces Cerevisiae ◽  
Dna Polymerase Δ ◽  
Repeat Units

AbstractThe POL30 gene of the yeast Saccharomyces cerevisiae encodes the proliferating cell nuclear antigen (PCNA), a protein required for processive DNA synthesis by DNA polymerase δ and ϵ. We examined the effects of the pol30-52 mutation on the stability of microsatellite (1- to 8-bp repeat units) and minisatellite (20-bp repeat units) DNA sequences. It had previously been shown that this mutation destabilizes dinucleotide repeats 150-fold and that this effect is primarily due to defects in DNA mismatch repair. From our analysis of the effects of pol30-52 on classes of repetitive DNA with longer repeat unit lengths, we conclude that this mutation may also elevate the rate of DNA polymerase slippage. The effect of pol30-52 on tracts of repetitive DNA with large repeat unit lengths was similar, but not identical, to that observed previously for pol3-t, a temperature-sensitive mutation affecting DNA polymerase δ. Strains with both pol30-52 and pol3-t mutations grew extremely slowly and had minisatellite mutation rates considerably greater than those observed in either single mutant strain.

scholarly journals Genes Involved in Sister Chromatid Separation and Segregation in the Budding Yeast Saccharomyces cerevisiae

Genetics ◽  
2001 ◽  
Vol 159 (2) ◽  
pp. 453-470
Author(s):  
Sue Biggins ◽  
Needhi Bhalla ◽  
Amy Chang ◽  
Dana L Smith ◽  
Andrew W Murray
Keyword(s):  
Chromosome Segregation ◽  
Sister Chromatid ◽  
Mitotic Spindle ◽  
Budding Yeast ◽  
Temperature Sensitive ◽  
Chromosome Behavior ◽  
Yeast Saccharomyces Cerevisiae ◽  
Sister Chromatids ◽  
Marked Chromosome ◽  
Sister Chromatid Separation

Abstract Accurate chromosome segregation requires the precise coordination of events during the cell cycle. Replicated sister chromatids are held together while they are properly attached to and aligned by the mitotic spindle at metaphase. At anaphase, the links between sisters must be promptly dissolved to allow the mitotic spindle to rapidly separate them to opposite poles. To isolate genes involved in chromosome behavior during mitosis, we microscopically screened a temperature-sensitive collection of budding yeast mutants that contain a GFP-marked chromosome. Nine LOC (loss of cohesion) complementation groups that do not segregate sister chromatids at anaphase were identified. We cloned the corresponding genes and performed secondary tests to determine their function in chromosome behavior. We determined that three LOC genes, PDS1, ESP1, and YCS4, are required for sister chromatid separation and three other LOC genes, CSE4, IPL1, and SMT3, are required for chromosome segregation. We isolated alleles of two genes involved in splicing, PRP16 and PRP19, which impair α-tubulin synthesis thus preventing spindle assembly, as well as an allele of CDC7 that is defective in DNA replication. We also report an initial characterization of phenotypes associated with the SMT3/SUMO gene and the isolation of WSS1, a high-copy smt3 suppressor.

scholarly journals SEN1, a positive effector of tRNA-splicing endonuclease in Saccharomyces cerevisiae.

1992 ◽  
Vol 12 (5) ◽  
pp. 2154-2164 ◽  
Author(s):  
D J DeMarini ◽  
M Winey ◽  
D Ursic ◽  
F Webb ◽  
M R Culbertson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Amino Acids ◽  
Amino Acid ◽  
Gene Product ◽  
Signal Sequence ◽  
Null Alleles ◽  
Nucleoside Triphosphate ◽  
Temperature Sensitive ◽  
Yeast Saccharomyces Cerevisiae ◽  
Amino Terminal

The SEN1 gene, which is essential for growth in the yeast Saccharomyces cerevisiae, is required for endonucleolytic cleavage of introns from all 10 families of precursor tRNAs. A mutation in SEN1 conferring temperature-sensitive lethality also causes in vivo accumulation of pre-tRNAs and a deficiency of in vitro endonuclease activity. Biochemical evidence suggests that the gene product may be one of several components of a nuclear-localized splicing complex. We have cloned the SEN1 gene and characterized the SEN1 mRNA, the SEN1 gene product, the temperature-sensitive sen1-1 mutation, and three SEN1 null alleles. The SEN1 gene corresponds to a 6,336-bp open reading frame coding for a 2,112-amino-acid protein (molecular mass, 239 kDa). Using antisera directed against the C-terminal end of SEN1, we detect a protein corresponding to the predicted molecular weight of SEN1. The SEN1 protein contains a leucine zipper motif, consensus elements for nucleoside triphosphate binding, and a potential nuclear localization signal sequence. The carboxy-terminal 1,214 amino acids of the SEN1 protein are essential for growth, whereas the amino-terminal 898 amino acids are dispensable. A sequence of approximately 500 amino acids located in the essential region of SEN1 has significant similarity to the yeast UPF1 gene product, which is involved in mRNA turnover, and the mouse Mov-10 gene product, whose function is unknown. The mutation that creates the temperature-sensitive sen1-1 allele is located within this 500-amino-acid region, and it causes a substitution for an amino acid that is conserved in all three proteins.

scholarly journals Osmotic stress and the yeast cytoskeleton: phenotype-specific suppression of an actin mutation.

1992 ◽  
Vol 118 (3) ◽  
pp. 561-571 ◽  
Author(s):  
S Chowdhury ◽  
K W Smith ◽  
M C Gustin
Keyword(s):  
Osmotic Stress ◽  
Actin Filament ◽  
Physiological Process ◽  
Actin Binding ◽  
Temperature Sensitive ◽  
Filamentous Actin ◽  
Cortical Actin ◽  
Yeast Saccharomyces Cerevisiae ◽  
Rhodamine Phalloidin ◽  
Diploid Cells

In the yeast Saccharomyces cerevisiae, actin filaments function to direct cell growth to the emerging bud. Yeast has a single essential actin gene, ACT1. Diploid cells containing a single copy of ACT1 are osmosensitive (Osms), i.e., they fail to grow in high osmolarity media (D. Shortle, unpublished observations cited by Novick, P., and D. Botstein. 1985. Cell. 40:415-426). This phenotype suggests that an underlying physiological process involving actin is osmosensitive. Here, we demonstrate that this physiological process is a rapid and reversible change in actin filament organization in cells exposed to osmotic stress. Filamentous actin was stained using rhodamine phalloidin. Increasing external osmolarity caused a rapid loss of actin filament cables, followed by a slower redistribution of cortical actin filament patches. In the recovery phase, cables and patches were restored to their original levels and locations. Strains containing an act1-1 mutation are both Osms and temperature-sensitive (Ts) (Novick and Botstein, 1985). To identify genes whose products functionally interact with actin in cellular responses to osmotic stress, we have isolated extragenic suppressors which revert only the Osms but not the Ts phenotype of an act1-1 mutant. These suppressors identify three genes, RAH1-RAH3. Morphological and genetic properties of a dominant suppressor mutation suggest that the product of the wild-type allele, RAH3+, is an actin-binding protein that interacts with actin to allow reassembly of the cytoskeleton following osmotic stress.

Electrical Properties of β-FeSi2 Thin Films on Insulating Substrates

2003 ◽  
Vol 796 ◽  
Author(s):  
Kensuke Akiyama ◽  
Takeshi Kimura ◽  
Shin Nishiyama ◽  
Takeo Hattori ◽  
Naoki Ohashi ◽  
...  
Keyword(s):  
Thin Films ◽  
Iron Silicide ◽  
Amorphous Film ◽  
Rf Magnetron Sputtering ◽  
Atomic Ratio ◽  
Similar Temperature Dependence ◽  
Electrical Conductivities ◽  
Similar Temperature ◽  
Band Conduction ◽  
Insulating Substrates

ABSTRACTIron silicide thin films were prepared on insulating substrates using RF magnetron sputtering method. Amorphous, polycrystalline and epitaxial β-FeSi2 were obtained on MgO(001), Al2O3(110) and Al2O3(001) substrates, respectively. Electrical conductivities of these films showed similar temperature dependence. Intrinsic band conduction and hopping conduction mechanism were predominant above and below 600K, respectively. The localized ordering in the polycrystalline and epitaxial films that controled the movement of carriers were as low as in the amorphous film. For the epitaxial β-FeSi2 film, electrical conductivity below 600K were affected by atomic ratio of silicon to iron (Si/Fe) in the films, because the localized ordering in the films decreased as Si/Fe atomic ratio decreased.

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