GSE158062
The Candida albicans MEKK Ste11 has both structural and enzymatic functions in opaque cells under pheromone stimulation
In this study, we investigated the relationships among the pheromone components to identify how the MAPK proteins are coordinated through a scaffold that lacks a key domain. An attempt to bypass other components by artificially driving either Cst5 or Ste11 to the plasma membrane suggested that the interactions among the proteins are more complex than the linear cascade characterized in S. cerevisiae. Moreover, the findings suggest the MAPK proteins Hst7 and Ste11 regulate the translocation of Cst5 within the cytoplasm. Indeed, it appears that Ste11 interacts with Cst5 to generate a complex node to tether the downstream kinases where Ste11 can both act enzymatically on Hst7 as well as make the MAPK Cek1 available for phosphorylation and activation by Hst7. Overall design: Liquid cultures of wild-type (WT) opaque cells (SN148aa) and opaque cells of ste11?/? were adjusted to OD600 at 0.8 in 10 mL liquid SC-GlcNac medium and incubated at room temperature for 6 hours with shaking at 220 rpm. The wild-type strain was cultivated in two tubes: untreated and treated with 10 µg/mL a-pheromone. The mutant strain was untreated. We conduted RNAseq profiling for 2 replicates in each condition (untreated WT , treated WT and untreated ste11?/?).