A ubiquitin-protein ligase(E3)mutation of Saccharomyces cerevisiae suppressed by co-overexpression of two ubiquitin-specific processing proteases

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To isolate mutations related to the ubiquitin system, I constructed a plasmid carrying the <i>YUH1</i> and <i>UBP1</i> genes (genes of ubiquitin-speeifie processing proteases) whose expressions were under the control of the galactose-inducible <i>GAL1-GAL10</i> promoter. Cells of a strain carrying the plasmid were mutagenized with ethyl methanesulfonate. One mutant, which showed galactose-dependent growth at a high temperature (37°C), was isolated from about 380,000 mutagenized colonies. The mutation responsible for galactose-dependent growth at 37°C was a single nuclear recessive mutation designated as <i>uby1-1</i>. <i>UBP1</i> and <i>YUH1</i> as well as the <i>GAL1-GAL10</i> promoter are required to suppress <i>uby1-1</i>. At the restrictive temperature, a <i>uby1-1</i> mutant did not arrest at a specific phase of the cell cycle, but still lost viability. Even at the permissive temperature (30°C), the <i>uby1-1</i> mutant grew somewhat slowly and showed pleiotropic phenotypes including hypersensitivity to stresses such as cadmium and canavanine, and sporulation defects. The genomic DNA fragments in a single-copy plasmid which complemented <i>uby1-1</i> were isolated. Chromosomal mapping, sequencing and subcloning analyses indicated that the gene complementing <i>uby1-1</i> is <i>RSP5</i>, which encodes a ubiquitin-protein ligase (E3) homologous to E6-AP (E6 associated protein). Deletion, complementation and linkage analyses revealed that <i>UBY1</i> and <i>RSP5</i> are the same gene. Therefore, the E3 protein encoded by <i>RSP5</i> (<i>UBY1</i>) is required for vegetative growth, sporulation and stress response. The present procedure using suppression by co-overexpression of two cloned genes will be useful to isolate mutations of related genes and to analyze biochemical pathways and gene-interactions.<br>

収録刊行物

  • Genes & genetic systems

    Genes & genetic systems 71(2), 75-83, 1996-04

    The Genetics Society of Japan

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各種コード

  • NII論文ID(NAID)
    10003759705
  • NII書誌ID(NCID)
    AA11077421
  • 本文言語コード
    ENG
  • 資料種別
    ART
  • ISSN
    13417568
  • NDL 記事登録ID
    4163710
  • NDL 雑誌分類
    ZR1(科学技術--生物学)
  • NDL 請求記号
    Z53-W539
  • データ提供元
    CJP書誌  NDL  J-STAGE 
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