Structurally unique interaction of RBD-like and PH domains is crucial for yeast pheromone signaling

Yerko, Volodymyr; Sulea, Traian; Ekiel, Irena; Harcus, Doreen; Baardsnes, Jason; Cygler, Mirosław; Whiteway, Malcolm; Wu, Cunle

doi:10.1091/mbc.e12-07-0516

Cited by 5 publications

(6 citation statements)

References 59 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Similarly, the strain STE11-CAAX/STE11-CAAX containing a membrane targeted Ste11 could not mate or form sexual projections in the absence of CST5 though overexpression of Ste11 had been reported to partially rescue the pheromone response defect of cst5Δ/Δ ( Yi et al 2011 ). In S. cerevisiae , hyperactivated Ste11 forms can significantly bypass the absence of the scaffold protein ( Stevenson et al 1992 ; Yerko et al 2013 ). Our findings demonstrated that tagging Cst5 and Ste11 to the plasma membrane was not enough to generate signaling to trigger pheromone responses in the absence of pheromone components due to indispensable interactions among the upstream and downstream components that appear to tightly regulate the activation of the MAP kinase cascade in C. albicans .…”

Section: Discussionmentioning

confidence: 99%

“…CaSte11 shares 59% sequence identity with ScSte11. The MEKK Ste11 has two N-terminal regulatory domains in both organisms, SAM and RBL, which in yeast interact with the adaptor protein Ste50 and the scaffold protein Ste5, respectively, to ensure the signal is properly related to the environmental cues ( Yerko et al 2013 ). In C. albicans , the C-terminal kinase domain of Ste11 is responsible for phosphorylation and docking of Hst7 ( Côte et al 2011 ).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Signal-mediated localization of Candida albicans pheromone response pathway components

Costa

Omran

Law

et al. 2020

Self Cite

View full text Add to dashboard Cite

Candida albicans opaque cells release pheromones to stimulate cells of opposite mating type to activate their pheromone response pathway. Although this fungal pathogen shares orthologous proteins involved in the process with Saccharomyces cerevisiae, the pathway in each organism has unique characteristics. We have used GFP-tagged fusion proteins to investigate the localization of the scaffold protein Cst5, as well as the MAP kinases Cek1 and Cek2, during pheromone response in C. albicans. In wild-type cells, pheromone treatment directed Cst5-GFP to surface puncta concentrated at the tips of mating projections. These puncta failed to form in cells defective in either the Gα or β subunits. However, they still formed in response to pheromone in cells missing Ste11, but with the puncta distributed around the cell periphery in the absence of mating projections. These puncta were absent from hst7Δ/Δ cells, but could be detected in the ste11Δ/Δ hst7Δ/Δ double mutant. Cek2-GFP showed a strong nuclear localization late in the response, consistent with a role in adaptation, while Cek1-GFP showed a weaker, but early increase in nuclear localization after pheromone treatment. Activation loop phosphorylation of both Cek1 and Cek2 required the presence of Ste11. In contrast to Cek2-GFP, which showed no localization signal in ste11Δ/Δ cells, Cek1-GFP showed enhanced nuclear localization that was pheromone independent in the ste11Δ/Δ mutant. The results are consistent with CaSte11 facilitating Hst7-mediated MAP kinase phosphorylation and also playing a potentially critical role in both MAP kinase and Cst5 scaffold localization.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Signal-mediated localization of Candida albicans pheromone response pathway components

Costa

Omran

Law

et al. 2020

Self Cite

View full text Add to dashboard Cite

show abstract

“…cerevisiae . The range of domains were determined as follow: ScSte11 SAM (aa 21–83), ScSte11 RBL (aa 116–236), ScSte11 kinase (aa 415–717) ( Yerko et al, 2013 ); KlSte11 SAM (aa 11–73), KlSte11 RBL (aa 107–228), KlSte11 kinase (aa 439–730); KmSte11 SAM (aa 11–73), KmSte11 RBL (aa 107–228), KmSte11 kinase (aa 445–735); ScSte7 kinase (aa 191–466) ( Won et al, 2011 ); KlSte7 kinase (aa 166–442); KmSte7 kinase (aa 155–431); ScFus3 kinase (aa 13–309); KlFus3 kinase (aa 13–327); KmFus3 kinase (aa 13–322); ScSte5 RING (aa 177–229), ScSte5 PH (aa 388–518), ScSte5 Fus3BD (aa 288–315), ScSte5 VWA (aa 593–786), ScSte5 Ste7BD (aa 744–895) ( Good et al, 2011 ); KlSte5 RING (aa 128–181), KlSte5 PH (aa 331–450), KlSte5 VWA (aa 498–683), KlSte5 Ste7BD (aa 647–746); KmSte5 RING (aa 136–189), KmSte5 PH (aa 345–464), KmSte5 VWA (aa 512–702), and KmSte5 Ste7BD (aa 660–763).…”

Section: Methodsmentioning

confidence: 99%

Correlation Between Improved Mating Efficiency and Weakened Scaffold-Kinase Interaction in the Mating Pheromone Response Pathway Revealed by Interspecies Complementation

et al. 2022

View full text Add to dashboard Cite

Scaffold protein Ste5 and associated kinases, including Ste11, Ste7, and Fus3, are core components of the mating pheromone pathway, which is required to induce a mating response. Orthologs of these proteins are widely present in fungi, but to which extent one protein can be replaced by its ortholog is less well understood. Here, interspecies complementation was carried out to evaluate the functional homology of Ste5 and associated kinases in Kluyveromyces lactis, K. marxianus, and Saccharomyces cerevisiae. These three species occupy important positions in the evolution of hemiascomycetes. Results indicated that Ste5 and associated kinases in K. lactis and K. marxianus could be functionally replaced by their orthologs to different extents. However, the extent of sequence identity, either between full-length proteins or between domains, did not necessarily indicate the extent of functional replaceability. For example, Ste5, the most unconserved protein in sequence, achieved the highest average functional replaceability. Notably, swapping Ste5 between K. lactis and K. marxianus significantly promoted mating in both species and the weakened interaction between the Ste5 and Ste7 might contribute to this phenotype. Consistently, chimeric Ste5 displaying a higher affinity for Ste7 decreased the mating efficiency, while chimeric Ste5 displaying a lower affinity for Ste7 improved the mating efficiency. Furthermore, the length of a negatively charged segment in the Ste7-binding domain of Ste5 was negatively correlated with the mating efficiency in K. lactis and K. marxianus. Extending the length of the segment in KlSte5 improved its interaction with Ste7 and that might contribute to the reduced mating efficiency. Our study suggested a novel role of Ste5-Ste7 interaction in the negative regulation of the pheromone pathway. Meanwhile, Ste5 mutants displaying improved mating efficiency facilitated the breeding and selection of Kluyveromyces strains for industrial applications.

show abstract

“…9A. To localize the Ahk1 binding domain in Ste11, we constructed a series of Ste11 deletion mutants that lacked, respectively, the sterile alpha motif (SAM) domain (27,28,49), the Rasbinding-domain-like (RBL) domain (50), and the autoinhibitory (AI) domain (Fig. 9A).…”

Section: Role Of the Cytoplasmic Domain Of Hkr1 In The Hog Pathwaymentioning

confidence: 99%

Scaffold Protein Ahk1, Which Associates with Hkr1, Sho1, Ste11, and Pbs2, Inhibits Cross Talk Signaling from the Hkr1 Osmosensor to the Kss1 Mitogen-Activated Protein Kinase

Nishimura

Yamamoto

Oyama

et al. 2016

Molecular and Cellular Biology

View full text Add to dashboard Cite

Structurally unique interaction of RBD-like and PH domains is crucial for yeast pheromone signaling

Cited by 5 publications

References 59 publications

Signal-mediated localization of Candida albicans pheromone response pathway components

Signal-mediated localization of Candida albicans pheromone response pathway components

Correlation Between Improved Mating Efficiency and Weakened Scaffold-Kinase Interaction in the Mating Pheromone Response Pathway Revealed by Interspecies Complementation

Scaffold Protein Ahk1, Which Associates with Hkr1, Sho1, Ste11, and Pbs2, Inhibits Cross Talk Signaling from the Hkr1 Osmosensor to the Kss1 Mitogen-Activated Protein Kinase

Contact Info

Product

Resources

About