2019 journal article

Systematic analysis of F-box proteins reveals a new branch of the yeast mating pathway

JOURNAL OF BIOLOGICAL CHEMISTRY, 294(40), 14717–14731.

By: N. Rangarajan*, C. Gordy*, L. Askew*, S. Bevill*, T. Elston*, B. Errede*, J. Hurst*, J. Kelley* ...

co-author countries: United States of America 🇺🇸
author keywords: G protein; mitogen-activated protein kinase (MAPK); yeast; ubiquitin; protein sorting; F-box; fungal mating; pheromone signaling
MeSH headings : Adaptor Proteins, Signal Transducing / genetics; Cell Cycle / genetics; Class III Phosphatidylinositol 3-Kinases / genetics; Endosomes / genetics; F-Box Proteins / chemistry; F-Box Proteins / genetics; GTP-Binding Protein alpha Subunits, Gq-G11 / genetics; GTP-Binding Protein beta Subunits / chemistry; GTP-Binding Protein beta Subunits / genetics; GTP-Binding Protein gamma Subunits / chemistry; GTP-Binding Protein gamma Subunits / genetics; Genes, Mating Type, Fungal / genetics; Morphogenesis / genetics; Pheromones / genetics; Pheromones / metabolism; Saccharomyces cerevisiae / genetics; Saccharomyces cerevisiae / physiology; Saccharomyces cerevisiae Proteins / genetics; Sequence Deletion / genetics; Signal Transduction; Transcription, Genetic; Vacuoles / genetics; Vacuoles / metabolism; cdc42 GTP-Binding Protein / genetics
Source: Web Of Science
Added: December 9, 2019

The mating pathway in yeast Saccharomyces cerevisiae has long been used to reveal new mechanisms of signal transduction. The pathway comprises a pheromone receptor, a heterotrimeric G protein, and intracellular effectors of morphogenesis and transcription. Polarized cell growth, in the direction of a potential mating partner, is accomplished by the G-protein βγ subunits and the small G-protein Cdc42. Transcription induction, needed for cell–cell fusion, is mediated by Gβγ and the mitogen-activated protein kinase (MAPK) scaffold protein Ste5. A potential third pathway is initiated by the G-protein α subunit Gpa1. Gpa1 signaling was shown previously to involve the F-box adaptor protein Dia2 and an endosomal effector protein, the phosphatidylinositol 3-kinase Vps34. Vps34 is also required for proper vacuolar sorting and autophagy. Here, using a panel of reporter assays, we demonstrate that mating pheromone stimulates vacuolar targeting of a cytoplasmic reporter protein and that this process depends on Vps34. Through a systematic analysis of F-box deletion mutants, we show that Dia2 is required to sustain pheromone-induced vacuolar targeting. We also found that other F-box proteins selectively regulate morphogenesis (Ydr306, renamed Pfu1) and transcription (Ucc1). These findings point to the existence of a new and distinct branch of the pheromone-signaling pathway, one that likely leads to vacuolar engulfment of cytoplasmic proteins and recycling of cellular contents in preparation for mating. The mating pathway in yeast Saccharomyces cerevisiae has long been used to reveal new mechanisms of signal transduction. The pathway comprises a pheromone receptor, a heterotrimeric G protein, and intracellular effectors of morphogenesis and transcription. Polarized cell growth, in the direction of a potential mating partner, is accomplished by the G-protein βγ subunits and the small G-protein Cdc42. Transcription induction, needed for cell–cell fusion, is mediated by Gβγ and the mitogen-activated protein kinase (MAPK) scaffold protein Ste5. A potential third pathway is initiated by the G-protein α subunit Gpa1. Gpa1 signaling was shown previously to involve the F-box adaptor protein Dia2 and an endosomal effector protein, the phosphatidylinositol 3-kinase Vps34. Vps34 is also required for proper vacuolar sorting and autophagy. Here, using a panel of reporter assays, we demonstrate that mating pheromone stimulates vacuolar targeting of a cytoplasmic reporter protein and that this process depends on Vps34. Through a systematic analysis of F-box deletion mutants, we show that Dia2 is required to sustain pheromone-induced vacuolar targeting. We also found that other F-box proteins selectively regulate morphogenesis (Ydr306, renamed Pfu1) and transcription (Ucc1). These findings point to the existence of a new and distinct branch of the pheromone-signaling pathway, one that likely leads to vacuolar engulfment of cytoplasmic proteins and recycling of cellular contents in preparation for mating.