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. 2016 Feb 8;202(4):1421–1435. doi: 10.1534/genetics.115.185207

Figure 2.

Figure 2

Kel1p overexpression suppresses the mating defect of fus2 mutations. (A) Suppressor plasmid pMR6441, containing KEL1 and REC104 ORFs, partially rescues the mating defect of Fus2p C-terminal mutations. fus2∆ (MY10904) strains already containing a plasmid with either fus2-L674A (pMR6501) or fus2-670UAG (pMR6775) were transformed with either pMR6441 or an empty 2μ plasmid (pRS425). These strains were mated to a fus1∆fus2∆ (JY429) for 3 hr at 30°. (B and C) KEL1 ORF is responsible for the suppression. (B) Two amino acids were inserted near the N termini of either KEL1 (pMR6730) or REC104 (pMR6731) in pMR6441 to create a stop codon and frameshift. Suppression was assessed via quantitative filter matings against a fus1∆fus2∆ (JY429) for 4 hr at 30°. (C) pMR6775 was transformed into a wild-type KEL1 strain (MY10904) as well as a strain where KEL1 was expressed under the control of the GAL1 promoter integrated at the KEL1 locus (MY13522). Suppression of fus2-670UAG was assessed via diploid formation. (D and E) High-copy KEL1 partially suppresses the mating defect of a complete fus2 deletion. (D) A fus2∆ strain (MY10904) was transformed with pMR6441 or an empty 2μ plasmid (pRS425). These strains were mated to a fus1∆fus2∆ (JY429) for 3 hr at 30°. (E) High-copy KEL1 suppresses C-terminal mutations better than the complete deletion. The same strains as in A and D were mated to a fus1∆fus2∆ (JY429) for 4 hr at 30° and suppression was assessed via diploid formation.