PMC

Signaling pathway in the regulation of gluconeogenesis by carbon source-responsive element-binding factors, Cat8p and Sip4p. Adapted from .

The expression profiles of genes CAT8, MIG1, and MLS1 in the BY and RM cocultures. The mRNA levels of genes CAT8 (A), MIG1 (B), and MLS1 (C) were quantified by TaqMan qRT-RCR using BY or RM allele-specific probes. The data from 2 replicates of the BY and RM cocultures are represented by solid and dotted lines and indicated as batch 1 and 2, respectively.

The growth and glucose concentration curves of the laboratory (BY) and wild (RM) strains. The cultures were started in yeast-rich growth media, YPAD, which has 2% glucose. The culture density is presented as the absorbance at 600 nm. At least 3 biological replicates were performed, and the growth and glucose concentration curves were similar to the ones shown here.

The expression profiles of genes CAT8, MIG1, and MLS1 in the BY–RM hybrid diploid strains YL123 and YL125. YL123 and YL125 were 2 independent reciprocal crosses that served as the replicates for the experiment. The mRNA levels of genes CAT8 (A), MIG1 (B), and MLS1 (C) were quantified by TaqMan qRT-RCR using BY or RM allele-specific probes. The data from strains YL123 and YL125 are represented by solid and dotted lines, respectively.

The expression profiles of genes CAT8 and MLS1 in the hybrid diploid strains YL135 and YL136 generated by mating BY and the swapped RM strain. YL135 and YL136 were 2 independent swapped hybrid clones that served as the replicates of the experiments. The mRNA levels of genes CAT8 (A) and MLS1 (B) were quantified by Taqman qRT-RCR using BY or RM allele-specific probes. The data from YL135 and YL136 are represented by solid and dotted lines, respectively.

The expression profiles of genes CAT8, MIG1, MLS1, and SNF1 in the BY and RM and YJM separated cultures. Each culture was harvested at different time points for RNA extraction, and the mRNA levels of genes CAT8 (A), MIG1 (B), and MLS1 (C), and SNF1 (D) were quantified by qRT-RCR. At least 3 biological replicates were performed, and the induction folds for CAT8, MIG1, and MLS1 were similar to the ones shown here.

The expression profiles of genes CAT8, MLS1, and MIG1 in separated cultures of BY, RM, and RM-swapped strains YL129 and YL130 in which we replaced the CAT8 promoter region of the RM allele by the one from the BY allele. YL129 and YL130 were 2 independent swapped clones that served as the replicates of the experiments. The mRNA levels of genes CAT8 (A), MLS1 (B), and MIG1 (C) were quantified by qRT-RCR. The data from the BY and RM strains are presented by solid lines, whereas those from YL129 and YL130 are presented by dotted lines.

Pyrosequencing results of MIG1, CAT8, and MLS1 from cocultures versus hybrids. The ratios of allele expression levels with the corresponding time pairs of coculture and hybrid are presented in the log scale. The diagonal line represents the 1-to-1 ratio, that is, BY/RM_hybrid = BY/RM_coculture. The vertical and horizontal bars on each dot represent the standard deviations from 2 biological and 2 technical repeats. (A) Pyrosequencing results of MIG1, CAT8, and MLS1 between strains. MIG1 is in red, CAT8 in green, and MLS1 in blue. The MIG1 results that are close to the vertical line are for YJM/RM and those near the diagonal line are for BY/RM or BY/YJM. The data points of CAT8 and MLS1 are for BY/RM (below the diagonal line) or YJM/RM (above the horizontal line). (B) Pyrosequencing results of MIG1 between the parental BY strain, the promoter-swapped strains, and the RM strain. The swapped strains were obtained by replacing the MIG1 promoter region in the BY strain that included 3 or 4 substitution sites with the same promoter region of the RM strain as described in Materials and Methods. The number “3” or “4” represents the number of key candidate nucleotides (see text) that were swapped. The data indicated in blue are for the parental BY and RM strains, whereas those indicated in orange are for the swapped strain with 3 substitution sites and the RM strain, and those indicated in red are for the swapped strain with 4 substitution sites and the RM strain.