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. 2012 Dec 11;7(12):e51551. doi: 10.1371/journal.pone.0051551

Figure 3. Yml081Wp regulates ALD gene expression and protein levels.

Figure 3

(A) Acetate biosynthesis pathway, adapted from Saint-Prix et al. [6]. (B) ALD gene expression was measured in mid-log phase wild-type and YML081W-null cells by quantitative PCR. Removal of YML081W resulted in a significant reduction in ALD4 and ALD6 mRNA levels. (C) ALD gene expression was measured in mid-log phase yeast cells overexpressing YML081W under the control of the PGK1 promoter, and compared to wild-type cells. YML081W-overexpressing cells produced significantly higher levels of ALD4 and ALD6 mRNA compared to wild-type cells. (D) ALD4 and ALD6 genes were tagged with the FLAG epitope at the C-terminus in wild-type, YML081W-null and YML081Woverexpressing strains. The cells were grown to mid-log phase, then harvested and lysed. The lysates were immunoblotted with an α-FLAG antibody to detect the levels of Ald4p-FLAG and Ald6p-FLAG protein. Cells lacking YML081W produced lower levels of FLAG-tagged Ald4p and Ald6p, compared to wild-type cells. Cells overexpressing YML081W produced significantly higher levels of Ald4p-FLAG protein compared to wild-type cells. ALD6p-FLAG levels did not appear to change significantly. Membrane staining shown below the immunoblots indicates the equivalance of total protein loading between lanes.