type by4742 cells  (Favorgen Biotech)

Journal: PLoS ONE

Article Title: Mmi1, the Yeast Homologue of Mammalian TCTP, Associates with Stress Granules in Heat-Shocked Cells and Modulates Proteasome Activity

doi: 10.1371/journal.pone.0077791

Figure Lengend Snippet: Yeast strains used in this study.

Article Snippet: An aliquot of 1 µg of the PCR product was transformed into BY4741 or BY4742 wild type cells, chromosomal integration was selected by plating the cells on YPD plates containing 200 µg/ml G418 (Geneticin, Sigma Aldrich, USA).

Techniques:

Journal: PLoS ONE

Article Title: Mmi1, the Yeast Homologue of Mammalian TCTP, Associates with Stress Granules in Heat-Shocked Cells and Modulates Proteasome Activity

doi: 10.1371/journal.pone.0077791

Figure Lengend Snippet: (A) Growth curves of BY4742 wild type (WT) cells as well as mmi1 Δ cells in the BY4742 strain (strain CRY1981) with and without the addition of 1 mM and 3 mM hydrogen peroxide. (B) Survival of WT and mmi1 Δ cells of both mating types (strains BY4741, BY4742, CRY1107, CRY1981) after a temperature shift from 30°C to 46°C for time periods of up to 100 min. Note the very marked increase of heat shock resistance of the deletion mutants. Error bars denote standard deviations of the mean obtained form 3 independent repeats of the experiment. (C) Changes of Mmi1-GFP distribution after a temperature shift from 30°C to 37°C, 40°C, 42°C and 46°C for 10 min each (strain CRY1838). Scale bar 4 µm. The figures show transfer of Mmi1 to part of the nuclear compartment at intermediate temperatures and transfer to both the nucleus and cytoplasmic granules upon 10 min heat shock at 46°C.

Article Snippet: An aliquot of 1 µg of the PCR product was transformed into BY4741 or BY4742 wild type cells, chromosomal integration was selected by plating the cells on YPD plates containing 200 µg/ml G418 (Geneticin, Sigma Aldrich, USA).

Techniques:

Journal: PLoS ONE

Article Title: Mmi1, the Yeast Homologue of Mammalian TCTP, Associates with Stress Granules in Heat-Shocked Cells and Modulates Proteasome Activity

doi: 10.1371/journal.pone.0077791

Figure Lengend Snippet: We compared the distribution of stress granules markers Pab1-GFP and Rpg1-RFP fusion proteins produced from sites on chromosomes in wild type (strain CRY527) and mmi1 Δ (strain CRY1060) cells. (A) Stress granule formation after robust heat shock was not influenced at all by the absence of Mmi1 in the mmi1 Δ deletion strain. (B) In the same strains that were used in (A) recovery from heat shock was observed for 60 min (shown) and 120 min. As shown, absence of Mmi1 had no influence on the dissolution of stress granules during recovery from the heat shock. Scale bar 4 µm.

Article Snippet: An aliquot of 1 µg of the PCR product was transformed into BY4741 or BY4742 wild type cells, chromosomal integration was selected by plating the cells on YPD plates containing 200 µg/ml G418 (Geneticin, Sigma Aldrich, USA).

Techniques: Produced

Journal: PLoS ONE

Article Title: Mmi1, the Yeast Homologue of Mammalian TCTP, Associates with Stress Granules in Heat-Shocked Cells and Modulates Proteasome Activity

doi: 10.1371/journal.pone.0077791

Figure Lengend Snippet: (A) Both fusion proteins Mmi1-GFP and Rpn1-RFP were expressed from the chromosomal sites (strain CRY1231) and co-localized in control (30°C) and 10 min heat-shocked cells (46°C). Control cells at 30°C displayed almost no overlaps of the two fusion proteins. This was confirmed by a negative value of the Pearsońs correlation coefficient (R r ). However, the cells heat-shocked at 46°C for 10 min displayed co-localization of both fusion proteins at the nuclear region. (B) In cells recovering from the heat stress, Mmi1 granules were dissolving during the time indicated whereas partial Mmi1-GFP location in the nuclear region remained detectable. However, decreased values of Rr in cells recovering from the heat shock for 60 min indicate continuous separation of the Mmi1-GFP and the Rpn1-RFP signals. Scale bar 4 µm. (C, D, E, F) We measured proteasomal proteolytic activity in low speed (50× g) supernatants prepared from cells either of the wild type strain (WT; strain CRY564) or the mmi1Δ strain (strain CRY1062), growing at 30°C or heat-shocked at 46°C for 10 min. Error bars indicate standard errors of eleven independent experiments. (C) A small but highly significant (p = 0.005) increase of 6.7% in proteasomal activity of the mmi1Δ strain was observed at 30°C. (D) Influence of the heat shock treatment on proteasomal activity in WT cells. A modest but significant (p = 0.003) increase of 7.6% in proteasomal activity was observed in WT cells. (E) Influence of the heat shock treatment on proteasomal activity in mmi1Δ cells . An 12.2% increase in proteasomal activity with high significance (p = 1. E-5) was observed. (F) Comparison of the WT and mmi1Δ strains after heat shock at 46°C. A large and significant (p = 0.007) increase of 11.8% in the proteasomal activity of the mmi1Δ strain was observed after heat-shock. We conclude that heat shock results in increase of the proteasomal activity and a presence of Mmi1 displays an inhibitory function in regulation of the proteasomal activity which is most pronounced after heat-shock.

Article Snippet: An aliquot of 1 µg of the PCR product was transformed into BY4741 or BY4742 wild type cells, chromosomal integration was selected by plating the cells on YPD plates containing 200 µg/ml G418 (Geneticin, Sigma Aldrich, USA).

Techniques: Activity Assay

Journal: FEMS yeast research

Article Title: End-of-life cell cycle arrest contributes to stochasticity of yeast replicative aging

doi: 10.1111/1567-1364.12030

Figure Lengend Snippet: Table 1

Article Snippet: Consistent with the results of Falcon and Aris( Falcon & Aris, 2003 ), we find that BY4742 wild type cells containing a CEN marked pMoBY vector are slightly short-lived relative to untransformed cells and also tend to arrest more frequently as budded cells ( ).

Techniques: