Fig 4.

Roles of Rsp5 WW domains and multiple PPxY proteins in Tat1 degradation in response to high pressure. (A) Cells of the wild-type, HPG1-1, rsp5-ww1, rsp5-ww2, rsp5-ww3, and bul1Δ bul2Δ strains were exposed to a pressure of 25 MPa for 3 h. (B) Cells of the wild type and of art single deletion mutants were exposed to a pressure of 25 MPa for 3 h. (C) (Left) Cells of the wild-type (BY4742), 9-arrestin (EN60), and 9-arrestin bul1Δ bul2Δ (EN67) strains were exposed to a pressure of 25 MPa for 3 h. Whole-cell extracts were prepared for Western blotting to detect the variant 3HA-Tat1 proteins. Adh1 was used as a loading control. (Right) Amounts of 3HA-Tat1 relative to that of Adh1 were calculated from the signals detected in an ImageQuant LAS4000 minisystem. Data are shown as means ± standard deviations for three independent experiments.