A positive feedback loop involving the Spa2 SHD domain contributes to focal polarization

Journal Article · 07 February 2022 · PLoS ONE

DOI: https://doi.org/10.1371/journal.pone.0263347 · OSTI ID:1844292

Lawson, Michael J.; Drawert, Brian; Petzold, Linda ; Yi, Tau-Mu ; Arkowitz, ed., Robert Alan

Focal polarization is necessary for finely arranged cell-cell interactions. The yeast mating projection, with its punctate polarisome, is a good model system for this process. We explored the critical role of the polarisome scaffold protein Spa2 during yeast mating with a hypothesis motivated by mathematical modeling and tested by in vivo experiments. Our simulations predicted that two positive feedback loops generate focal polarization, including a novel feedback pathway involving the N-terminal domain of Spa2. We characterized the latter using loss-of-function and gain-of-function mutants. The N-terminal region contains a Spa2 Homology Domain (SHD) which is conserved from yeast to humans, and when mutated largely reproduced the spa2Δ phenotype. Our work together with published data show that the SHD domain recruits Msb3/4 that stimulates Sec4-mediated transport of Bud6 to the polarisome. There, Bud6 activates Bni1-catalyzed actin cable formation, recruiting more Spa2 and completing the positive feedback loop. We demonstrate that disrupting this loop at any point results in morphological defects. Gain-of-function perturbations partially restored focal polarization in a spa2 loss-of-function mutant without restoring localization of upstream components, thus supporting the pathway order. Thus, we have collected data consistent with a novel positive feedback loop that contributes to focal polarization during pheromone-induced polarization in yeast.

• Research Organization: Univ. of California, Santa Barbara, CA (United States)
• Sponsoring Organization: National Institute of Biomedical Imaging and Bioengineering (NIBIB); National Institute of General Medical Sciences (NIGMS); National Institutes of Health (NIH); National Science Foundation (NSF); US Army Research Office (ARO); USDOE; USDOE Office of Science (SC)
• Grant/Contract Number: SC0008975
• OSTI ID: 1844292
• Journal Information: PLoS ONE, Journal Name: PLoS ONE Journal Issue: 2 Vol. 17; ISSN 1932-6203
• Publisher: Public Library of Science (PLoS)Copyright Statement
• Country of Publication: United States
• Language: English