Interactions Between Positive and Negative Regulators of GCN4 Controlling Gene Expression and Entry Into the Yeast Cell Cycle

Satoshi Harashima ¹, Ernest M. Hannig ¹, and Alan G. Hinnebusch ¹

¹ Laboratory of Molecular Genetics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892

The GCN4 gene encodes a transcriptional activator in yeast whose expression is regulated at the translational level in response to amino acid availability. gcn3 mutations block derepression of GCN4 expression in starvation conditions. gcd1 and gcd12 mutations restore derepression of GCN4 expression in gcn3 deletion mutants, suggesting that GCN3 positively regulates GCN4 indirectly by antagonism of these GCD functions. gcd1 and gcd12 mutations also lead to temperature-sensitive arrest in the G₁ phase of the cell cycle in gcn3 deletion mutants. The GCN3 allele completely suppresses both derepression of GCN4 expression and the temperature-sensitive growth conferred by gcd12 mutations and partially suppresses these phenotypes in gcd1 mutants. This suggests that the GCN3 product can promote or provide GCD function in nonstarvation conditions even though it opposes GCD function when cells are starved for amino acids. The gcn3-102 allele is completely defective for positive regulation of GCN4 expression; however, it mimics GCN3 in suppressing gcd1 and gcd12 mutations and thus retains the ability to restore GCD function in nonstarvation conditions. These data suggest that GCN3, GCD1 and GCD12 have closely related functions required for regulation of GCN4 expression and entry into the cell cycle. We suggest that GCN3 antagonizes the regulatory functions of GCD1 and GCD12 in starvation conditions either by competing with these factors for the same sites of action or by modifying their structures by physical interaction.

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