FIGURE 2: Mec1 promotes the carbon source dependent regulation of Rnr3. (A) WT, sml1Δ, and mec1-kd sml1Δ cells were grown to mid-log phase in YPD (“D”) or YPG (“G”). WCE samples were prepared and subjected to Western blot analysis using α-Rnr1 and α-GFP antibodies. Tubulin was used as a loading control. (B, C) WT and mec1-4 cells were grown to mid log phase in YPD (D) or YPG (G) at 25°C, a permissive temperature for mec1-4 [8]. WCE samples were prepared and subjected to α-Rnr1 and α-GFP Western blot analysis. “G →D”: Cells grown to mid-log phase in YPG were collected and released into YPD and incubated at three different temperatures, 25°C, 30°C, and 37°C. WCE samples were prepared at 3- and 22-hours after the medium switch and subjected to Western blot analysis using α-Rnr1 and α-GFP antibodies. Tubulin was used as a loading control.

8. Corcoles-Saez I, Dong K, Waskiewicz E, Costanzo M, Boone C, and Cha RS (2018). Essential Function of Mec1, the Budding Yeast ATM/ATR Checkpoint-Response Kinase, in Protein Homeostasis. Dev Cell 46: 495–503. doi: 10.1016/j.devcel.2018.07.011