Identification of dosage-sensitive genes in Saccharomyces cerevisiae using the genetic tug-of-war method

Koji Makanae, Reiko Kintaka, Takashi Makino, Hiroaki Kitano, Hisao Moriya

    Research output: Contribution to journalArticlepeer-review

    60 Citations (Scopus)

    Abstract

    Gene overexpression beyond a permissible limit causes defects in cellular functions. However, the permissible limits of most genes are unclear. Previously, we developed a genetic method designated genetic tug-of-war (gTOW) to measure the copy number limit of overexpression of a target gene. In the current study, we applied gTOW to the analysis of all protein-coding genes in the budding yeast Saccharomyces cerevisiae. We showed that the yeast cellular system was robust against an increase in the copy number by up to 100 copies in >80% of the genes. After frameshift and segmentation analyses, we isolated 115 dosage-sensitive genes (DSGs) with copy number limits of 10 or less. DSGs contained a significant number of genes involved in cytoskeletal organization and intracellular transport. DSGs tended to be highly expressed and to encode protein complex members. We demonstrated that the protein burden caused the dosage sensitivity of highly expressed genes using a gTOW experiment in which the open reading frame was replaced with GFP. Dosage sensitivities of some DSGs were rescued by the simultaneous increase in the copy numbers of partner genes, indicating that stoichiometric imbalances among complexes cause dosage sensitivity. The results obtained in this study will provide basic knowledge about the physiology of chromosomal abnormalities and the evolution of chromosomal composition.

    Original languageEnglish
    Pages (from-to)300-311
    Number of pages12
    JournalGenome Research
    Volume23
    Issue number2
    DOIs
    Publication statusPublished - 2013 Feb

    ASJC Scopus subject areas

    • Genetics
    • Genetics(clinical)

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