Role of the N-terminal region of Rap1p in the transcriptional activation of glycolytic genes in Saccharomyces cerevisiae

Authors

  • Takayuki Mizuno,

    1. Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki 305-8566, Japan
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  • Tomoko Kishimoto,

    1. Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki 305-8566, Japan
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  • Tomoko Shinzato,

    1. Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki 305-8566, Japan
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  • Robin Haw,

    1. Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki 305-8566, Japan
    Current affiliation:
    1. Banting & Best Department of Medical Research, University of Toronto, Toronto, Ontario, M5G 1L6, Canada.
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  • Alistair Chambers,

    1. Institute of Genetics, University of Nottingham, Queen's Medical Centre, Nottingham NG7 2UH, UK
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  • Jason Wood,

    1. Cancer Center, Harvard Medical School, Boston, MA 02115, USA
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  • David Sinclair,

    1. Cancer Center, Harvard Medical School, Boston, MA 02115, USA
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  • Hiroshi Uemura

    Corresponding author
    1. Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki 305-8566, Japan
    • Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), AIST Tsukuba Central 6 Higashi 1-1-1, Tsukuba, Ibaraki, 305-8566, Japan.
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Abstract

In the yeast two-hybrid system, the N-terminal region of Rap1p was shown to interact with Gcr1p and Gcr2p. Disruption of gcr1 and/or gcr2 in the two-hybrid reporter strain demonstrated that the interaction with Gcr1p does not require Gcr2p, whereas the interaction with Gcr2p is mediated through Gcr1p. Deletion of the N-terminal region of Rap1p alone did not show a growth phenotype, but a growth defect was observed when this mutation was combined with a gcr2 deletion. The poor growth of the gcr1 null mutant was not affected further by the N-terminal deletion of Rap1p, but the growth of gcr1 strains with mutations in the DNA binding region of Gcr1p was affected by the removal of the N-terminal region of Rap1p. These results suggest that one function of the N-terminal region of Rap1p, presumably the BRCT domain, is to facilitate the binding of Gcr1p to the promoter by a protein–protein interaction. Copyright © 2004 John Wiley & Sons, Ltd.

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