Banuelos MG~Gharakhanian E, 2010

Pubmed ID 20043226
Title Genomic analysis of severe hypersensitivity to hygromycin B reveals linkage to vacuolar defects and new vacuolar gene functions in Saccharomyces cerevisiae.
Authors M G Banuelos, D E Moreno, D K Olson, Q Nguyen, F Ricarte, C R Aguilera-Sandoval, Editte Gharakhanian
Abstract The vacuole of Saccharomyces cerevisiae has been a seminal model for studies of lysosomal trafficking, biogenesis, and function. Several yeast mutants defective in such vacuolar events have been unable to grow at low levels of hygromycin B, an aminoglycoside antibiotic. We hypothesized that such severe hypersensitivity to hygromycin B (hhy) is linked to vacuolar defects and performed a genomic screen for the phenotype using a haploid deletion strain library of non-essential genes. Fourteen HHY genes were initially identified and were subjected to bioinformatics analyses. The uncovered hhy mutants were experimentally characterized with respect to vesicular trafficking, vacuole morphology, and growth under various stress and drug conditions. The combination of bioinformatics analyses and phenotypic characterizations implicate defects in vesicular trafficking, vacuole fusion/fission, or vacuole function in all hhy mutants. The collection was enriched for sensitivity to monensin, indicative of vacuolar trafficking defects. Additionally, all hhy mutants showed severe sensitivities to rapamycin and caffeine, suggestive of TOR kinase pathway defects. Our experimental results also establish a new role in vacuolar and vesicular functions for two genes: PAF1, encoding a RNAP II-associated protein required for expression of cell cycle-regulated genes, and TPD3, encoding the regulatory subunit of protein phosphatase 2A. Thus, our results support linkage between severe hypersensitivity to hygromycin B and vacuolar defects.
Citation Curr. Genet. 2010; 56:121-37

Datasets

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Papers Phenotype Conditions Collection Tested mutants Data Details
Banuelos MG~Gharakhanian E, 2010 growth (spot assay) hygromycin B [25 ug/ml] hap alpha ~4,828 Discrete

Curation history

Data

March 26, 2014 Data waiting for tested.
Jan. 28, 2015 Data to load.
Dec. 9, 2016 Data loaded.

Tested strains

March 26, 2014 Tested strains requested.
Nov. 11, 2014 Tested strains requested.
Jan. 28, 2015 Tested strains abandoned.