Figure 3.

slx5-K suppressor alleles are hypomorphic. (A) Strain PSY3884 (slx5∆) was transformed with the indicated SLX5 allele in a LEU2/CEN/ARS plasmid. Transformants were subjected to 10-fold serial dilutions, and pinned onto selective medium in the presence or absence of 0.1 M HU. (B) Strain JMY2462 (sgs1∆ slx5∆ plus pJM500 [SGS1/ADE3/URA3/CEN]) was treated as in (A) except that transformants were pinned onto selective medium or SC medium containing FOA. (C) Strain JMY3694 (slx5∆ smt3-3KR plus pJM7496 [SLX5/SMT3/ADE3/URA3/CEN]) was treated as in (B). (D) Whole-cell extracts were prepared under denaturing conditions from strains with the indicated SLX5 genotype. Extracts were subjected to SDS-PAGE and immunoblotted with anti-Smt3 antibody. Shown is the high molecular-weight region of the blot including the stacking gel. The lower panel shows a portion of the membrane stained with ponceau S as loading control. (E) Strain PSY3868 (ulp2∆ slx5∆ + pMS7481 [ULP2/ADE3/URA3/CEN]) was treated as in (B) except cells were pinned onto medium lacking leucine in the presence or absence of FOA. (F) Strain PSY3866 (siz1∆ siz2∆ slx5∆ + pJM7494 [SLX5/ADE3/URA3/CEN]) was treated as in (B) except that transformants were streaked onto SC medium containing FOA.