Mutual suppression of phenotypes in the gas1 sas3 double mutant.

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Mutual suppression of phenotypes in the gas1 sas3 double mutant. Mutual suppression of phenotypes in the gas1 sas3 double mutant. (A) Deletion of SAS3 rescues gas1 temperature sensitivity and silencing defects at the telomere and rDNA array but not CFW sensitivity. In turn, deletion of GAS1 restores HM silencing in sas3 to wild-type levels. The sir2 mutant is included as a positive control for disruption of silencing. Top panel: Serial dilutions of wild type (LPY4924), sir2 (LPY5035), sas3 (LPY19731), gas1 (LPY19773), and gas1 sas3 (LPY16444) were plated on SC at 30° and 37°, SC with 5-FOA (TELVR::URA3) or SC −Trp (hml::TRP1). Middle panel: Serial dilutions of wild type (LPY2444), sir2 (LPY2447), sas3 (LPY17686), gas1 (LPY10074), and gas1 sas3 (LPY17685) were plated on SC or SC −Ura (RDN::Ty-1-mURA3) at 30°. Bottom panel: wild type (LPY5), sas3 (LPY8256), gas1 (LPY10129), and gas1 sas3 (LPY17520) were plated on either SC or SC with 10 μg/ml CFW. (B) NuA3 complex with subunits analyzed herein shaded green (adapted from Lafon et al. 2007). (C) Deletion of YNG1 does not have synthetic interactions with gas1. Serial dilutions of wild type (LPY6285), yng1 (LPY5526), gas1 (LPY9820), and gas1 yng1 (LPY16997) were plated on SC at either 30° or 37°. (D) Analysis of GAS1, GCN5, and SAS3 reveals distinct and opposing outcomes for synthetic interactions. Serial dilutions of wild type (LPY5), gcn5 (LPY8242), sas3 (LPY16039), gas1 (LPY10129), gas1 gcn5 + p-GCN5, URA3 (LPY16736), gas1 sas3 (LPY19823), and gas1 gcn5 sas3 + p-GCN5, URA3 (LPY19101) were plated on SC or SC with 5-FOA, to select against p-GCN5, URA3, at 30° with and without 1 M sorbitol. Moriah Eustice, and Lorraine Pillus Genetics 2014;196:1029-1039 Copyright © 2014 by the Genetics Society of America