Homing at an extragenic locus mediated by VDE (PI-SceI) in Saccharomyces cerevisiae

Authors

  • Satoru Nogami,

    1. Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Bldg. FSB-101, 5-1-5 Kashiwanoha, Kashiwa, Chiba Prefecture, 277-8562 Japan
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  • Tomoyuki Fukuda,

    1. Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Bldg. FSB-101, 5-1-5 Kashiwanoha, Kashiwa, Chiba Prefecture, 277-8562 Japan
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  • Yuri Nagai,

    1. Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Bldg. FSB-101, 5-1-5 Kashiwanoha, Kashiwa, Chiba Prefecture, 277-8562 Japan
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  • Shizu Yabe,

    1. Department of Biological Sciences, Graduate School of Sciences, University of Tokyo, Tokyo 113-0033, Japan
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  • Masako Sugiura,

    1. Department of Biological Sciences, Graduate School of Sciences, University of Tokyo, Tokyo 113-0033, Japan
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  • Ryuta Mizutani,

    1. Faculty of Pharmaceutical Sciences, University of Tokyo, Tokyo 113-0033, Japan
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  • Yoshinori Satow,

    1. Faculty of Pharmaceutical Sciences, University of Tokyo, Tokyo 113-0033, Japan
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  • Yasuhiro Anraku,

    1. Department of Biosciences, Teikyo University of Science and Technology, Yatsuzawa 2525, Uenohara-machi, Yamanashi 409-0193, Japan
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  • Yoshikazu Ohya

    Corresponding author
    1. Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Bldg. FSB-101, 5-1-5 Kashiwanoha, Kashiwa, Chiba Prefecture, 277-8562 Japan
    • Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Bldg. FSB-101, 5-1-5 Kashiwanoha, Kashiwa, Chiba Prefecture, 277-8562 Japan.
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Abstract

PI-SceI (VDE), a homing endonuclease with protein splicing activity, is a genomic parasite in the VMA1 gene of Saccharomyces cerevisiae. In a heterozygous diploid of the VDE-less VMA1 allele and a VDE-containing VMA1 allele, VDE specifically cleaves its recognition sequence (VRS) in the VDE-less VMA1 allele at meiosis, followed by ‘homing’, i.e. a conversion to a VDE-containing allele. We found that upon VDE expression, homing of a marker gene at an extragenic locus occurs only when a 45 bp element containing the VRS is inserted at its allelic site, while mutants of VDE with no endonuclease activity lack authentic extragenic homing activity. Thus, both the VRS and VDE are required for homing. Insertion of the VRS in a homozygous diploid significantly lowered the spore germination ability, indicating that a template for gene repair at its allelic locus is essential for efficient homing and survival of yeast cells. Copyright © 2002 John Wiley & Sons, Ltd.

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