The isolation of biologically active mating pheromone, a-factor, from the yeast, Saccharomyces cerevisiae.
Haploid cell-types of baker's yeast, Saccharomyces cerevisiae, secrete pheromones which are essential for conjugation. Recently a putative structure for the elusive a-factor pheromone has been reported. In this report we present a procedure to obtain a-factor from batch cultures of cells using hydrophobic Amberlite XAD-2 resin in the growth medium with subsequent differential washings of the resin with organic solvents. We have determined the biological activity of the a-factor preparation by verifying that there is an increase in transcription of the a-factor receptor gene, STE3, by Northern analysis of STE3 mRNA before and after exposure of the appropriate cell type to a-factor. Furthermore, a beta-galactosidase assay of the putative receptor gene fused to the lacZ gene, coding for beta-galactosidase (STE3-lacZ), was done to quantify the biological activity of the a-factor.
Proteau
G
G
Department of Zoology, Erindale College, University of Toronto, Mississauga, Ontario, Canada.
Gelaznikas
R
R
Merante
F
F
D000595:Amino Acid Sequence; D007475:Ion Exchange Resins; D000072235:Mating Factor; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D010675:Pheromones / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D011137:Polystyrenes; D011956:Receptors, Cell Surface / Q000096:biosynthesis / Q000378:metabolism; D051964:Receptors, Mating Factor; D018000:Receptors, Peptide; D012441:Saccharomyces cerevisiae / Q000032:analysis*; D014157:Transcription Factors
D007475:Ion Exchange Resins; D010455:Peptides; D010675:Pheromones; D011137:Polystyrenes; D011956:Receptors, Cell Surface; D051964:Receptors, Mating Factor; D018000:Receptors, Peptide; D014157:Transcription Factors; D000072235:Mating Factor; C003771:Amberlite XAD-2 resin