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Suppressors of MPS3-G186K toxicity.

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posted on 17.11.2011, 00:22 by Jennifer M. Friederichs, Suman Ghosh, Christine J. Smoyer, Scott McCroskey, Brandon D. Miller, Kyle J. Weaver, Kym M. Delventhal, Jay Unruh, Brian D. Slaughter, Sue L. Jaspersen

(A) Mutants from the yeast deletion collection were transformed with pURA3-GAL-MPS3-G186K, and transformants were tested for their ability to grow on SC-URA+2% galactose at for 3 d at 30°C. Possible hits were then further examined by western blotting to ensure that Mps3-G186K was expressed at high levels, similar to the wild-type control. Shown here are the hits that both rescued the growth defect of MPS3-G186K and expressed wild-type levels of Mps3-G186K. A full list of genes recovered in this screen is presented in Table S2. Also shown are the wild-type strain and the gal4Δ mutant, which we isolated in the primary screen but eliminated in the secondary screen because it does not express MPS3-G186K. Hits are organized by function using GoSlim analysis. In some cases, we isolated a mutant that deletes part of an overlapping gene, which is indicated below in parenthesis. (B) The indicated genes were deleted in GAL-MPS3-G186K (SLJ1797) and growth was tested in a serial dilution assay at 30°C on YPD (glucose) for 2 d and YPGR (galactose) for 3 d.