Nielsen, Ida5; Bentsen, Iben Bach6; Andersen, Anni Hangaard6; Gasser, Susan M.4; Bjergbæk, Lotte6
1 Department of Molecular Biology and Genetics, Science and Technology, Aarhus University2 Department of Molecular Biology and Genetics - Genome stability and technology, Department of Molecular Biology and Genetics, Science and Technology, Aarhus University3 Department of Biomedicine - Forskning og uddannelse, Syd, Department of Biomedicine, Health, Aarhus University4 Friedrich Miescher Institute for Biomedical Research, Basel5 Department of Biomedicine - Forskning og uddannelse, Syd, Department of Biomedicine, Health, Aarhus University6 Department of Molecular Biology and Genetics - Genome stability and technology, Department of Molecular Biology and Genetics, Science and Technology, Aarhus University
The conserved family of RecQ DNA helicases consists of caretaker tumour suppressors, that defend genome integrity by acting on several pathways of DNA repair that maintain genome stability. In budding yeast, Sgs1 is the sole RecQ helicase and it has been implicated in checkpoint responses, replisome stability and dissolution of double Holliday junctions during homologous recombination. In this study we investigate a possible genetic interaction between SGS1 and RAD9 in the cellular response to methyl methane sulphonate (MMS) induced damage and compare this with the genetic interaction between SGS1 and RAD24. The Rad9 protein, an adaptor for effector kinase activation, plays well-characterized roles in the DNA damage checkpoint response, whereas Rad24 is characterized as a sensor protein also in the DNA damage checkpoint response. Here we unveil novel insights into the cellular response to MMS-induced damage. Specifically, we show a strong synergistic functionality between SGS1 and RAD9 for recovery from MMS induced damage and for suppression of gross chromosomal rearrangements, which is not the case for SGS1 and RAD24. Intriguingly, it is a Rad53 independent function of Rad9, which becomes crucial for genome maintenance in the absence of Sgs1. Despite this, our dissection of the MMS checkpoint response reveals parallel, but unequal pathways for Rad53 activation and highlights significant differences between MMS- and hydroxyurea (HU)-induced checkpoint responses with relation to the requirement of the Sgs1 interacting partner Topoisomerase III (Top3). Thus, whereas earlier studies have documented a Top3-independent role of Sgs1 for an HU-induced checkpoint response, we show here that upon MMS treatment, Sgs1 and Top3 together define a minor but parallel pathway to that of Rad9.