Increased Expression of Hsp70 for Resistance to Deuterium Oxide in a Yeast Mutant Cell Line

Unno, Keiko; Kishido, Takahiro; Morioka, Mizue; Okada, Shoji; Oku, Naoto

doi:10.1248/bpb.26.799

Cited by 9 publications

(5 citation statements)

References 30 publications

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“…However, inducing molecular chaperones through a heat shock prior to incubation with D2O did not protect cells from the D2Oinduced growth retardation. This indicates that activation of the stress response pathway is not tuned to counter the adverse effects of the solvent exchange, although it has been reported that overexpression of Hsp70 may protect budding yeast cells from D2O [55]. In line with chaperones being unable to protect fission yeast cells against D2O, we were unable to adapt S. pombe to growth at higher D2O concentrations and we did not identify any chaperone mutants as hypersensitive or resistant to D2O.…”

Section: Discussionmentioning

confidence: 58%

“…This indicates that the activation of this stress response pathway is not tuned to counter the adverse effects of the solvent exchange; however it has been reported that the overexpression of Hsp70 may protect budding yeast cells from D 2 O. 62 In line with chaperones being unable to protect fission yeast cells against D 2 O, we were unable to adapt S. pombe to growth at higher D 2 O concentrations, and we did not identify any chaperone mutants as hypersensitive or resistant to D 2 O. Conversely, it was not surprising to find that pretreating cells with D 2 O led to an increased tolerance toward heat and oxidizing conditions, which is most likely caused by the D 2 O-induced activation of the heatshock response. This activated stress response and slow growth phenotypes are probably also connected with our observation that exposure to D 2 O caused a strong decrease in the expression of several tRNAs.…”

Section: ■ Discussionmentioning

confidence: 64%

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Mutations in a Single Signaling Pathway Allow Cell Growth in Heavy Water

et al. 2020

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Since life is completely dependent on water, it is difficult to gauge the impact of solvent change. To analyze the role of water as a solvent in biology, we replaced water with heavy water (D2O), and investigated the biological effects by a wide range of techniques, using the fission yeast Schizosaccharomyces pombe as model organism. We show that high concentrations of D2O lead to altered glucose metabolism, growth retardation, and inhibition of meiosis. However, mitosis and overall cell viability were only slightly affected. After prolonged incubation in D2O, cells displayed gross morphological changes, thickened cell walls as well as aberrant septa and cytoskeletal organization. RNA sequencing revealed that D2O causes a strong downregulation of most tRNAs and triggers activation of the general stress response pathway. Genetic screens identified several D2O sensitive mutants, while mutants compromised in the cell integrity pathway, including the protein kinase genes pmk1, mkh1, pek1 and pck2, that control cell wall biogenesis, were more tolerant to D2O. We speculate that D2O affects the phospholipid membrane or cell wall glycans causing an activation of the cell integrity pathway. In conclusion, the effects of solvent replacement are pleiotropic but the D2O-triggered activation of the cell integrity pathway and subsequent increased deposition of cell wall material and septation problems appear most critical for the cell growth defects..

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Section: Discussionmentioning

confidence: 58%

Section: ■ Discussionmentioning

confidence: 64%

Mutations in a Single Signaling Pathway Allow Cell Growth in Heavy Water

et al. 2020

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“…A slightly higher induction of the HSP gene in the D2Ocontaining medium was found for all treatments tested. This is most probably due to an increased general stress caused by the D2O-containing medium rather than due to any specific stress caused by 1 O2 (47). Earlier expression analysis with DNA microarrays indicates that only a few genes are induced by 1 O2 in C. reinhardtii, suggesting the involvement of a specific mechanism in the response to 1 O2 (unpublished data).…”

Section: Discussionmentioning

confidence: 91%

“…We studied the effect of D 2 O-supplemented growth medium on the expression of several stress genes, known to be induced by RB and/or NR and thus analyzed, whether 1 O 2 may be involved in the response of these genes upon exposure to these photosensitizers. First, a culture of strain cw 15 arg 7 mt - was pregrown for 3−4 d in TAP medium containing 50% D 2 O to adapt the cells and allow the cells to overcome the D 2 O-caused stress ( , ). Subsequently, the cells were exposed to a low concentration of the photosensitizer RB (0.3 μM) or NR (1 μM) and to 2 mM H 2 O 2 , inducing an oxidative stress not involving 1 O 2 .…”

Section: Resultsmentioning

confidence: 99%

Photosensitizers Neutral Red (Type I) and Rose Bengal (Type II) Cause Light-Dependent Toxicity in Chlamydomonas reinhardtii and Induce the Gpxh Gene via Increased Singlet Oxygen Formation

Fischer

Krieger‐Liszkay

Eggen

2004

Environ. Sci. Technol.

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The connection between the mode of toxic action and the genetic response caused by the type I photosensitizer and photosynthesis inhibitor neutral red (NR) and the type II photosensitizer rose bengal (RB) was investigated in the green alga Chlamydomonas reinhardtii. For both photosensitizers, a light intensity-dependent increase in toxicity and expression of the glutathione peroxidase homologous gene (Gpxh) was found. The toxicity of RB was reduced by the singlet oxygen (1O2) quenchers 1,4-diazabicyclo[2.2.2]octane and L-histidine, and the RB-induced Gpxh expression was stimulated in deuterium oxide-supplemented growth medium. These observations clearly indicate the involvement of 1O2 in both toxicity and the genetic response caused by RB. NR up-regulated the expression of typical oxidative and general stress response genes, probably by a type I mechanism, and also strongly induced the Gpxh expression. The stimulating effect of deuterium oxide in the growth medium suggested the involvement of 1O2 also in the NR-induced response. Indeed, an increased 1O2 formation was detected with EPR-spin trapping in NR-treated spinach thylakoids. However, none of the 102 quenchers could reduce the light-dependent toxicity of NR in C. reinhardtii, indicating that NR has a different mode of toxic action than RB.

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“…Several studies have reported on the biological effects of deuterium on microorganisms but the underlying mechanisms of growth reduction remain poorly understood. In yeast which was adapted to growth in deuterated media an increased production of the heat shock protein, Hsp70 was observed 5 . In bacteria it was reported that deuterium in the growth medium increases catalase activity and induce effects similar to that of osmotic shock 6 .…”

Section: Introductionmentioning

confidence: 99%

Strain improvement of Escherichia coli K-12 for recombinant production of deuterated proteins

Kelpšas

Wachenfeldt

2019

Sci Rep

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Deuterium isotope labelling is important for structural biology methods such as neutron protein crystallography, nuclear magnetic resonance and small angle neutron scattering studies of proteins. Deuterium is a natural low abundance stable hydrogen isotope that in high concentrations negatively affect growth of cells. The generation time for Escherichia coli K-12 in deuterated medium is substantially increased compared to cells grown in hydrogenated (protiated) medium. By using a mutagenesis plasmid based approach we have isolated an E. coli strain derived from E. coli K-12 substrain MG1655 that show increased fitness in deuterium based growth media, without general adaptation to media components. By whole-genome sequencing we identified the genomic changes in the obtained strain and show that it can be used for recombinant production of perdeuterated proteins in amounts typically needed for structural biology studies.

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Increased Expression of Hsp70 for Resistance to Deuterium Oxide in a Yeast Mutant Cell Line

Cited by 9 publications

References 30 publications

Mutations in a Single Signaling Pathway Allow Cell Growth in Heavy Water

Mutations in a Single Signaling Pathway Allow Cell Growth in Heavy Water

Photosensitizers Neutral Red (Type I) and Rose Bengal (Type II) Cause Light-Dependent Toxicity in Chlamydomonas reinhardtii and Induce the Gpxh Gene via Increased Singlet Oxygen Formation

Strain improvement of Escherichia coli K-12 for recombinant production of deuterated proteins

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