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. 2013 Dec 13;196(2):427–438. doi: 10.1534/genetics.113.160093

Figure 5.

Figure 5

Mutations in tV(UAC)D inhibit expression and function of Atg31 protein. (A) Western blot analysis was performed on wild-type and tDNA mutants containing an ATG31–9X-myc tag allele to determine whether the extended transcript affects translation of Atg31p. Compared to wild type, tDNA mutants showed significantly reduced expression of Atg31p. No other bands were observed on the Western blot, demonstrating that the extended transcript does not lead to production of an extended polypeptide. Strains used were DDY5012 (wt), DDY5014 (tdnaΔ), DDY5018 (B-boxΔ), and DDY5020 (B-box mut). Extracts were prepared from cells grown in YPD media. (B) Inhibition of autophagy induction in yeast expressing the extended transcript as measured by the Pho8Δ60 alkaline phosphatase assay. Strains used were: DDY5051 (wt control), DDY5072 (tdnaΔ), DDY5078 (B-boxΔ), DDY5081 (B-box mut), DDY5044 (atg8Δ), and DDY5046 (atg31Δ). (C) Production of the extended transcript is associated with reduced survival under nitrogen starvation conditions. Strains used were: DDY5012 (wt), DDY5081 (B-box mut), and DDY4764 (atg31Δ).