YOR387C

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Systematic name YOR387C
Gene name
Aliases
Feature type ORF, Uncharacterized
Coordinates Chr XV:1070241..1069621
Primary SGDID S000005914


Description of YOR387C: Putative protein of unknown function; regulated by the metal-responsive Aft1p transcription factor; highly inducible in zinc-depleted conditions; localizes to the soluble fraction[1][2][3]




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Community Commentary

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This gene is part of the UW-Stout Orphan Gene Project. Learn more here.

Growth Curve

YOR387C-water.png

In a BY4735 background, knocking out YOR387C does not seem to have much effect on the strain's growth rate. In this experiment, the BY4735 strain's doubling time was 162 minutes, while the YOR387C knock-out strain's doubling time was 137 minutes. (These doubling times are the means of three experiments.)

Hydroxychloroquine

YOR387C.png

Under normal conditions, the BY4753 doubling time is 144 minutes, while that of YOR387C is 158 minutes. On average, the YOR387C lag phase was slightly longer than that of BY4753 under normal conditions. When grown in an environment with hydroxychloroquine, YOR387C doubled in 249 minutes, while BY4753 doubled in 276 minutes. This shows that hydroxychloroquine inhibits BY4753 slightly more than YOR387C. (These doubling times and curves are the means of three experiments.)


Salt Concentration (NaCl)

NaCl-BY4735vYOR387C-Graph.JPG


0mM NaCl conc. BY4735 strain's doubling time: 169 minutes

0mM NaCl conc. YOR387C strain's doubling time: 129 minutes

750mM NaCl conc. BY4735 strain's doubling time: 294 minutes

750mM NaCl conc. YOR387C strain's doubling time: 282 minutes


The graph above shows the growth rate for the previously listed strains and the relative level of NaCl concentration. Knocking out the gene positively influences the growth rate. Comparing the YOR387C doubling times in 0mM and 750mM NaCl, there is a large effect on the growth rate. This effect negatively impacts the knockout strain's (YOR387C) growth rate.


Caffeine Group 1

gene1vswildgene123.jpg

Caffeine had an effect on the YOR387C strain; it had slowed the growth rate of the yeast. The doubling times are as follows:

Doubling time of YOR387C with caffeine: 458 minutes

Doubling time of YOR387C without caffeine: 122 minutes

Doubling time of BY4735 with caffeine: 138 minutes

Doubling time of BY4735 without caffeine: 187 minutes


Competative Co-culture

YOR387CCompetetiveCoCulture.jpg

These are the results of a competitive co-culture protocol. The knockout strain was grown in a culture to test fitness against a green, fluorescent wild-type strain. The percentage of analyzed cells in the culture measured was those displaying green fluorescence. In theory, this should mean if a knockout has reduced the fitness of a strain of yeast, the fluorescent wild-type strain would have a higher percentage than the knockout strain. If the knockout does not decrease fitness, they would be roughly equal. This may not be so in results. Sources of error may include contamination and human error.

The results here are not expected in theory. Test 1 was more likely than not contaminated with a faster growing organism than yeast that did not possess fluorescence. Strain 2 gave an expected result of about a 1:1 ratio that suggests the knockout of YOR378C in yeast does not reduced its fitness in a fair, competitive scenario.

Ammonium Sulfate

TNKO1.jpg

YOR387C gene does not show any major sensitivities to nitrogen starvation. The growth rate is 7.3% slower in the negative control than that of the 5g/ml treatment. The wild strains comparative growth rate was 17.8% slower. This is not much different in the scope of the data, but it could indicate that YOR387C is slightly less sensitive to nitrogen fixation than the wild strain.

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UV Light

Knock-out Strain 1

Results:

  • Experiment 1:0sec=342 colonies, 600sec=13 colonies.
  • Experiment 2:0sec=222 colonies, 600sec=42 colonies.

Interpretation: The two 0sec plates are on top and the 600sec plates are on the bottom. Assuming the plates had around the same amount of yeast cells, the UV light killed a majority of the yeast cells meaning it's possible that the knocked-out gene had something to do with UV resistance/DNA repair.

References

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  1. Higgins VJ, et al. (2003) Application of genome-wide expression analysis to identify molecular markers useful in monitoring industrial fermentations. Appl Environ Microbiol 69(12):7535-40 SGD PMID 14660410
  2. Rutherford JC, et al. (2003) Aft1p and Aft2p mediate iron-responsive gene expression in yeast through related promoter elements. J Biol Chem 278(30):27636-43 SGD PMID 12756250
  3. Terashima H, et al. (2002) Sequence-based approach for identification of cell wall proteins in Saccharomyces cerevisiae. Curr Genet 40(5):311-6 SGD PMID 11935221

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