Current Genetics

, Volume 56, Issue 2, pp 121–137

Genomic analysis of severe hypersensitivity to hygromycin B reveals linkage to vacuolar defects and new vacuolar gene functions in Saccharomyces cerevisiae

  • M. G. Banuelos
  • D. E. Moreno
  • D. K. Olson
  • Q. Nguyen
  • F. Ricarte
  • C. R. Aguilera-Sandoval
  • Editte Gharakhanian
Research Article

DOI: 10.1007/s00294-009-0285-3

Cite this article as:
Banuelos, M.G., Moreno, D.E., Olson, D.K. et al. Curr Genet (2010) 56: 121. doi:10.1007/s00294-009-0285-3
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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.

Keywords

Yeast vacuole Lysosome Hygromycin B Vesicular trafficking 

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • M. G. Banuelos
    • 1
  • D. E. Moreno
    • 1
  • D. K. Olson
    • 1
  • Q. Nguyen
    • 1
  • F. Ricarte
    • 1
  • C. R. Aguilera-Sandoval
    • 1
  • Editte Gharakhanian
    • 1
  1. 1.Department of Biological SciencesCalifornia State University at Long BeachLong BeachUSA

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