YLR006C

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Systematic name YLR006C
Gene name SSK1
Aliases
Feature type ORF, Verified
Coordinates Chr XII:163893..161755
Primary SGDID S000003996


Description of YLR006C: Cytoplasmic response regulator; part of a two-component signal transducer that mediates osmosensing via a phosphorelay mechanism; required for mitophagy; dephosphorylated form is degraded by the ubiquitin-proteasome system; potential Cdc28p substrate[1][2][3][4]




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Community Commentary

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Protein Details

Protein Modification

Modification(s): Phosphorylation

Identified as an efficient substrate of Clb2-Cdk1-as1 in a screen of a proteomic GST-fusion library. [4] [5]

Acids

UW-Stout/Acids_SP23

File:MEL control v.s knockout -2

Caffeine

UW-Stout/Caffeine SP23

KnockoutGeneControl.jpg
KnockoutGene2.jpg

Analysis


Glucose Sensitivity

Protocol

[1]


TableGlucose.jpg

Analysis

The graph displayed above shows a viability assay of SSK1 knockout gene as compared to BY4735 (wild type) and other knockout strains. We found that there was about 20% less cell death then in the control wild type. It was surprising to find that by removing this gene there was an increase in cell viability. Further research may be conducted to test this slight resistance to all cell death, but more importantly the resistance to glucose-induced apoptosis as noted in the wild type BY4735 for more conclusive results.

Isopropanol

UW-Stout/Isopropanol_SP23

Picture11.jpg

Analysis

These results were what was expected with only a little bit of difference. The first trial and third trials were close and there was less growth than the control which could have been because the knockout genes were more sensitive to the isopropanol. The second trial gave different results in which there was more growth than the control which is the opposite of the first and third. This could have been caused by an error because there was consistency in the other two.


Heat Shock

UW-Stout/Heat_shock_SP23

SSK1 Yeast Colonies at 40 degrees Celsius
Time (sec) Colonies
0 24
15 14
45 8
75 9
105 10
135 15
Wild Type Yeast Colonies at 40 degrees Celsius
Time (sec) Colonies
0 24
15 20
45 17
75 16
105 18
135 14

DKATrial2Yeast.jpg

KDA SSK1.jpg


Analysis

The general trend of our data yielded a negative relationship between time of heat shock. About half of the yeast cells died off in a matter of 135 seconds at 40 degrees Celsius. The SSK1 yeast were dying off at a very comparable rate over time compared to the Wild Yeast strain. This means that the gene that was knocked out must not have a correlation with heat shock.

Bases

EMYeast2.jpg

Analysis Yeast 2, A4 A5 A6, showed almost no growth compared to the Wild Type yeast, A1 A2 A3. This knockout gene likely did not benefit the resistance to basic pH changes in the environment.

Protocol- UW-Stout/Bases SP23

References

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  1. Mao K, et al. (2011) Two MAPK-signaling pathways are required for mitophagy in Saccharomyces cerevisiae. J Cell Biol 193(4):755-67 SGD PMID 21576396
  2. Posas F, et al. (1996) Yeast HOG1 MAP kinase cascade is regulated by a multistep phosphorelay mechanism in the SLN1-YPD1-SSK1 two-component osmosensor. Cell 86(6):865-75 SGD PMID 8808622
  3. Sato N, et al. (2003) Phosphorelay-regulated degradation of the yeast Ssk1p response regulator by the ubiquitin-proteasome system. Mol Cell Biol 23(18):6662-71 SGD PMID 12944490
  4. 4.0 4.1 Ubersax JA, et al. (2003) Targets of the cyclin-dependent kinase Cdk1. Nature 425(6960):859-64 SGD PMID 14574415 Cite error: Invalid <ref> tag; name "S000074306" defined multiple times with different content
  5. submitted by Jeff Ubersax on 2004-01-29

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