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Chemical-genetic profile analysis of five inhibitory compounds in yeast.

Alamgir M, Erukova V, Jessulat M, Azizi A, Golshani A - BMC Chem Biol (2010)

Bottom Line: Further investigations were carried out to assess the activity of three profiled genes in the process of protein biosynthesis using relative fitness of double mutants and other genetic assays.Our follow-up investigations into the activity of three profiled genes in the process of protein biosynthesis provided further evidence concerning the usefulness of chemical-genetic analyses for annotating gene functions.We termed these genes TAE2, TAE3 and TAE4 for translation associated elements 2-4.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biology, Carleton University, 1125 Colonel By Drive, Ottawa, K1 S 5B6, ON, Canada. ashkan_golshani@carleton.ca.

ABSTRACT

Background: Chemical-genetic profiling of inhibitory compounds can lead to identification of their modes of action. These profiles can help elucidate the complex interactions between small bioactive compounds and the cell machinery, and explain putative gene function(s).

Results: Colony size reduction was used to investigate the chemical-genetic profile of cycloheximide, 3-amino-1,2,4-triazole, paromomycin, streptomycin and neomycin in the yeast Saccharomyces cerevisiae. These compounds target the process of protein biosynthesis. More than 70,000 strains were analyzed from the array of gene deletion mutant yeast strains. As expected, the overall profiles of the tested compounds were similar, with deletions for genes involved in protein biosynthesis being the major category followed by metabolism. This implies that novel genes involved in protein biosynthesis could be identified from these profiles. Further investigations were carried out to assess the activity of three profiled genes in the process of protein biosynthesis using relative fitness of double mutants and other genetic assays.

Conclusion: Chemical-genetic profiles provide insight into the molecular mechanism(s) of the examined compounds by elucidating their potential primary and secondary cellular target sites. Our follow-up investigations into the activity of three profiled genes in the process of protein biosynthesis provided further evidence concerning the usefulness of chemical-genetic analyses for annotating gene functions. We termed these genes TAE2, TAE3 and TAE4 for translation associated elements 2-4.

No MeSH data available.


Related in: MedlinePlus

Strain sensitivity to different translation-inhibitory drugs. Wild type (WT) or gene deletion mutant strains (yploo9cΔ, yil137cΔ, ypl183w-aΔ, ydr056cCΔ and yjr111cΔ) were serially diluted to 10-3 to 10-6 and spotted on solid medium with sub-inhibitory concentrations of cycloheximide, paromomycin, 3-AT, streptomycin and neomycin as indicated, or without drugs (control). The plates were incubated at 30°C for 1-2 days. Deletion of ypl009c confers increased sensitivity to cycloheximide; yil137c and ypl183w-a to 3-AT, ydr056c to streptomycin and neomycin, and yjr111c to streptomycin.
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Figure 2: Strain sensitivity to different translation-inhibitory drugs. Wild type (WT) or gene deletion mutant strains (yploo9cΔ, yil137cΔ, ypl183w-aΔ, ydr056cCΔ and yjr111cΔ) were serially diluted to 10-3 to 10-6 and spotted on solid medium with sub-inhibitory concentrations of cycloheximide, paromomycin, 3-AT, streptomycin and neomycin as indicated, or without drugs (control). The plates were incubated at 30°C for 1-2 days. Deletion of ypl009c confers increased sensitivity to cycloheximide; yil137c and ypl183w-a to 3-AT, ydr056c to streptomycin and neomycin, and yjr111c to streptomycin.

Mentions: To investigate the accuracy of our large-scale approach to detect drug sensitive mutants, five deletion strains were selected and subjected to spot test analysis (Figure 2). This analysis confirmed that deletion of YPL009C confers increased sensitivity to cycloheximide, deletion of YDR056C increases sensitivity to streptomycin and neomycin, deletion of YJR111C increases sensitivity to streptomycin, and deletions of YIL137C and YPL183W-A increase sensitivity to 3-AT. These results are in agreement with the large-scale analysis and confirm that this approach can identify strains that are sensitive to the drugs used in this study.


Chemical-genetic profile analysis of five inhibitory compounds in yeast.

Alamgir M, Erukova V, Jessulat M, Azizi A, Golshani A - BMC Chem Biol (2010)

Strain sensitivity to different translation-inhibitory drugs. Wild type (WT) or gene deletion mutant strains (yploo9cΔ, yil137cΔ, ypl183w-aΔ, ydr056cCΔ and yjr111cΔ) were serially diluted to 10-3 to 10-6 and spotted on solid medium with sub-inhibitory concentrations of cycloheximide, paromomycin, 3-AT, streptomycin and neomycin as indicated, or without drugs (control). The plates were incubated at 30°C for 1-2 days. Deletion of ypl009c confers increased sensitivity to cycloheximide; yil137c and ypl183w-a to 3-AT, ydr056c to streptomycin and neomycin, and yjr111c to streptomycin.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2925817&req=5

Figure 2: Strain sensitivity to different translation-inhibitory drugs. Wild type (WT) or gene deletion mutant strains (yploo9cΔ, yil137cΔ, ypl183w-aΔ, ydr056cCΔ and yjr111cΔ) were serially diluted to 10-3 to 10-6 and spotted on solid medium with sub-inhibitory concentrations of cycloheximide, paromomycin, 3-AT, streptomycin and neomycin as indicated, or without drugs (control). The plates were incubated at 30°C for 1-2 days. Deletion of ypl009c confers increased sensitivity to cycloheximide; yil137c and ypl183w-a to 3-AT, ydr056c to streptomycin and neomycin, and yjr111c to streptomycin.
Mentions: To investigate the accuracy of our large-scale approach to detect drug sensitive mutants, five deletion strains were selected and subjected to spot test analysis (Figure 2). This analysis confirmed that deletion of YPL009C confers increased sensitivity to cycloheximide, deletion of YDR056C increases sensitivity to streptomycin and neomycin, deletion of YJR111C increases sensitivity to streptomycin, and deletions of YIL137C and YPL183W-A increase sensitivity to 3-AT. These results are in agreement with the large-scale analysis and confirm that this approach can identify strains that are sensitive to the drugs used in this study.

Bottom Line: Further investigations were carried out to assess the activity of three profiled genes in the process of protein biosynthesis using relative fitness of double mutants and other genetic assays.Our follow-up investigations into the activity of three profiled genes in the process of protein biosynthesis provided further evidence concerning the usefulness of chemical-genetic analyses for annotating gene functions.We termed these genes TAE2, TAE3 and TAE4 for translation associated elements 2-4.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biology, Carleton University, 1125 Colonel By Drive, Ottawa, K1 S 5B6, ON, Canada. ashkan_golshani@carleton.ca.

ABSTRACT

Background: Chemical-genetic profiling of inhibitory compounds can lead to identification of their modes of action. These profiles can help elucidate the complex interactions between small bioactive compounds and the cell machinery, and explain putative gene function(s).

Results: Colony size reduction was used to investigate the chemical-genetic profile of cycloheximide, 3-amino-1,2,4-triazole, paromomycin, streptomycin and neomycin in the yeast Saccharomyces cerevisiae. These compounds target the process of protein biosynthesis. More than 70,000 strains were analyzed from the array of gene deletion mutant yeast strains. As expected, the overall profiles of the tested compounds were similar, with deletions for genes involved in protein biosynthesis being the major category followed by metabolism. This implies that novel genes involved in protein biosynthesis could be identified from these profiles. Further investigations were carried out to assess the activity of three profiled genes in the process of protein biosynthesis using relative fitness of double mutants and other genetic assays.

Conclusion: Chemical-genetic profiles provide insight into the molecular mechanism(s) of the examined compounds by elucidating their potential primary and secondary cellular target sites. Our follow-up investigations into the activity of three profiled genes in the process of protein biosynthesis provided further evidence concerning the usefulness of chemical-genetic analyses for annotating gene functions. We termed these genes TAE2, TAE3 and TAE4 for translation associated elements 2-4.

No MeSH data available.


Related in: MedlinePlus