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Amino acids can induce yeast cell adhesion but how amino acids are sensed and signal the modulation of the  FLO  adhesion genes is not clear. We discovered that the budding yeast  Saccharomyces cerevisiae  CEN.PK evolved invasive growth ability under prolonged nitrogen limitation. Such invasive mutants were used to identify amino acid transporters as regulators of  FLO11  and invasive growth. One invasive mutant had elevated levels of  FLO11  mRNA and a Q320STOP mutation in the  SFL1  gene that encodes a protein kinase A pathway regulated repressor of  FLO11 . Glutamine-transporter genes  DIP5  and  GNP1  were essential for  FLO11  expression, invasive growth and biofilm formation in this mutant. Invasive growth relied on known regulators of  FLO11  and the Ssy1-Ptr3-Ssy5 complex that controls  DIP5  and  GNP1 , suggesting that Dip5 and Gnp1 operates downstream of the Ssy1-Ptr3-Ssy5 complex for regulation of  FLO11  expression in a protein kinase A dependent manner. The role of Dip5 and Gnp1 appears to be conserved in the  S. cerevisiae  strain ∑1278b since the  dip5 gnp1  ∑1278b mutant showed no invasive phenotype.  Secondly, the amino acid transporter gene  GAP1  was found to influence invasive growth through  FLO11  as well as other  FLO  genes. Cells carrying a dominant loss-of-function  PTR3 647::CWNKNPLSSIN   allele had increased transcription of the adhesion genes  FLO1, 5 ,  9 ,  10 ,  11  and the amino acid transporter gene  GAP1 . Deletion of  GAP1  caused loss of  FLO11  expression and invasive growth. However, deletions of  FLO11  and genes encoding components of the mitogen-activated protein kinase pathway or the protein kinase A pathway were not sufficient to abolish invasive growth, suggesting involvement of other  FLO  genes and alternative pathways. Increased intracellular amino acid pools in the  PTR3 647::CWNKNPLSSIN  -containing strain opens the possibility that Gap1 regulates the  FLO  genes through alteration of the amino acid pool sizes.

Amino Acid Transporter Genes Are Essential for FLO11-Dependent and FLO11-Independent Biofilm Formation and Invasive Growth in Saccharomyces cerevisiae