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Distinct associations of the Saccharomyces cerevisiae Rad9 protein link Mac1-regulated transcription to DNA repair

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Abstract

While it is known that ScRad9 DNA damage checkpoint protein is recruited to damaged DNA by recognizing specific histone modifications, here we report a different way of Rad9 recruitment on chromatin under non DNA damaging conditions. We found Rad9 to bind directly with the copper-modulated transcriptional activator Mac1, suppressing both its DNA binding and transactivation functions. Rad9 was recruited to active Mac1-target promoters (CTR1, FRE1) and along CTR1 coding region following the association pattern of RNA polymerase (Pol) II. Hir1 histone chaperone also interacted directly with Rad9 and was partly required for its localization throughout CTR1 gene. Moreover, Mac1-dependent transcriptional initiation was necessary and sufficient for Rad9 recruitment to the heterologous ACT1 coding region. In addition to Rad9, Rad53 kinase also localized to CTR1 coding region in a Rad9-dependent manner. Our data provide an example of a yeast DNA-binding transcriptional activator that interacts directly with a DNA damage checkpoint protein in vivo and is functionally restrained by this protein, suggesting a new role for Rad9 in connecting factors of the transcription machinery with the DNA repair pathway under unchallenged conditions.

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Acknowledgments

We thank Iannis Talianidis and the late George Thireos for materials and helpful suggestions, Dennis Winge for Mac1up-expressing plasmids, the late Yannis Papanikolau and Androniki Kretsovali for advice on protein methodologies, Ioannis Kagiampakis and Christos Andreadis for communicating experimental data, and George A. Garinis for critical reading of the manuscript. Dedicated to the late Alexandros Argyrokastritis.

Funding

This work was supported by the Greek Ministry of Development-GSRT (IMBB funding and PENED grant 01ED119) and Greek Ministry of Education (PYTHAGORAS grant 89184).

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Authors and Affiliations

  1. Department of Biology, University of Crete, Vasilika Vouton, P.O.Box 2208, 710 03, Heraklion, Crete, Greece

    Kalliopi Gkouskou & Despina Alexandraki

  2. Institute of Molecular Biology & Biotechnology, Foundation for Research and Technology—Hellas, N. Plastira 100, 70013, Heraklion, Crete, Greece

    George S. Fragiadakis, Alexandra Voutsina & Despina Alexandraki

  3. Embiodiagnostics, Heraklion, Crete, Greece

    Kalliopi Gkouskou

  4. Laboratory of Tumor Cell Biology, School of Medicine, University of Crete, P.O. Box 2208, 710 03, Heraklion, Crete, Greece

    Alexandra Voutsina

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  1. Kalliopi Gkouskou
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  2. George S. Fragiadakis
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  3. Alexandra Voutsina
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  4. Despina Alexandraki
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Correspondence to Despina Alexandraki.

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Communicated by M. Kupiec.

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Gkouskou, K., Fragiadakis, G.S., Voutsina, A. et al. Distinct associations of the Saccharomyces cerevisiae Rad9 protein link Mac1-regulated transcription to DNA repair. Curr Genet 66, 531–548 (2020). https://doi.org/10.1007/s00294-019-01047-w

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  • DOI: https://doi.org/10.1007/s00294-019-01047-w

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