G3: Genes|Genomes|Genetics Early Online, published on May 27, 2016 as doi:10.1534/g3.116.029520 1 Saccharomyces cerevisiae Tti2 regulates PIKK proteins and stress response Kyle S. Hoffman,* Martin L. Duennwald, † Jim Karagiannis,‡ Julie Genereaux,* Alexander S. McCarton,* Christopher J. Brandl*,1 *Department of Biochemistry, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, Ontario, Canada N6A5C1, †Department of Pathology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, Ontario, Canada N6A5C1, ‡Department of Biology, The University of Western Ontario, London, Ontario, Canada N6A5B7 © The Author(s) 2013. Published by the Genetics Society of America. 2 Running title: Tti2 is required for stress response Key words: TTT complex, PIKK proteins, heat shock response, chaperone, protein expression Corresponding Author Information: Dr. Christopher Brandl, Department of Biochemistry, The University of Western Ontario, 1151 Richmond St., London, Ontario, Canada N6A5C1 Tel: 519-661-2111 ext. 86857 email:
[email protected] 3 ABSTRACT The TTT complex is composed of the three essential proteins Tel2, Tti1 and Tti2. The complex is required to maintain the steady state levels of the phosphatidylinositol 3-kinase-related kinase (PIKK) proteins including mTOR, ATM/Tel1, ATR/Mec1, and TRRAP/Tra1, all of which serve as regulators of critical cell signaling pathways. Due to their association with heat shock proteins, and with newly synthesized PIKK peptides, components of the TTT complex may act as co-chaperones. Here, we analyze the consequences of depleting the cellular level of Tti2 in Saccharomyces cerevisiae. We show that yeast expressing low levels of Tti2 are viable under optimal growth conditions, but the cells are sensitive to a number of stress conditions that involve PIKK pathways.