Ypk1蛋白激酶信号通路被重新连接,对白色念珠菌的生存能力不是必需的。

IF 4.5 2区 生物学 Q1 Agricultural and Biological Sciences PLoS Genetics Pub Date : 2023-08-10 eCollection Date: 2023-08-01 DOI:10.1371/journal.pgen.1010890
Bernardo Ramírez-Zavala, Ines Krüger, Andreas Wollner, Sonja Schwanfelder, Joachim Morschhäuser
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引用次数: 0

摘要

蛋白激酶是几乎所有控制细胞活动的信号通路的中心成分。在模式生物酿酒酵母中,控制膜脂质稳态的同源蛋白激酶Ypk1和Ypk2对生存能力至关重要,先前的研究强烈表明,在致病性白色念珠菌中,它们的单一同源蛋白激酶也存在这种情况。在这里,使用FLP介导的诱导型基因缺失,我们发现白色念珠菌ypk1Δ突变体是可行的,但生长缓慢,这解释了以前未能获得无效突变体的原因。突变体的表型分析表明,Ypk1在调节鞘脂生物合成和细胞膜脂质不对称方面的功能是保守的,但Ypk1缺失的后果比酿酒酵母中的要轻。突变研究表明,其激活环中高度保守的PDK1磷酸化位点T548对Ypk1的功能至关重要,而C末端的TORC2磷酸化位点S687和T705对Ypk1-依赖性膜胁迫抗性至关重要。出乎意料的是,在非表达条件下,白色念珠菌的正常生长不需要Pkh1/Pkh2的单一白色念珠菌同源物Pkh1,它介导酿酒酵母PDK1位点的Ypk1磷酸化,并且在Pkh1Δ突变体中,T548处的Ypkl磷酸化仅略有减少。我们发现另一种蛋白激酶,Pkh3,其在酿酒酵母中的直系同源物不能取代Pkh1/2,与Pkh1冗余作用以激活白色念珠菌中的Ypk1。在缺乏单独的Pkh3的细胞中没有观察到表型效应,但pkh1ΔPkh3Δ双突变体具有严重的生长缺陷,并且T548处的Ypk1磷酸化被完全消除。这些结果表明,Ypk1对白色念珠菌的生存能力不是必需的,尽管其功能通常是保守的,但Ypk1信号通路在这种致病酵母中被重新连接,并包括一种新的上游激酶,通过PDK1位点的磷酸化激活Ypk1。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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The Ypk1 protein kinase signaling pathway is rewired and not essential for viability in Candida albicans.

Protein kinases are central components of almost all signaling pathways that control cellular activities. In the model organism Saccharomyces cerevisiae, the paralogous protein kinases Ypk1 and Ypk2, which control membrane lipid homeostasis, are essential for viability, and previous studies strongly indicated that this is also the case for their single ortholog Ypk1 in the pathogenic yeast Candida albicans. Here, using FLP-mediated inducible gene deletion, we reveal that C. albicans ypk1Δ mutants are viable but slow-growing, explaining prior failures to obtain null mutants. Phenotypic analyses of the mutants showed that the functions of Ypk1 in regulating sphingolipid biosynthesis and cell membrane lipid asymmetry are conserved, but the consequences of YPK1 deletion are milder than in S. cerevisiae. Mutational studies demonstrated that the highly conserved PDK1 phosphorylation site T548 in its activation loop is essential for Ypk1 function, whereas the TORC2 phosphorylation sites S687 and T705 at the C-terminus are important for Ypk1-dependent resistance to membrane stress. Unexpectedly, Pkh1, the single C. albicans orthologue of Pkh1/Pkh2, which mediate Ypk1 phosphorylation at the PDK1 site in S. cerevisiae, was not required for normal growth of C. albicans under nonstressed conditions, and Ypk1 phosphorylation at T548 was only slightly reduced in pkh1Δ mutants. We found that another protein kinase, Pkh3, whose ortholog in S. cerevisiae cannot substitute Pkh1/2, acts redundantly with Pkh1 to activate Ypk1 in C. albicans. No phenotypic effects were observed in cells lacking Pkh3 alone, but pkh1Δ pkh3Δ double mutants had a severe growth defect and Ypk1 phosphorylation at T548 was completely abolished. These results establish that Ypk1 is not essential for viability in C. albicans and that, despite its generally conserved function, the Ypk1 signaling pathway is rewired in this pathogenic yeast and includes a novel upstream kinase to activate Ypk1 by phosphorylation at the PDK1 site.

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来源期刊
PLoS Genetics
PLoS Genetics 生物-遗传学
CiteScore
8.10
自引率
2.20%
发文量
438
审稿时长
1 months
期刊介绍: PLOS Genetics is run by an international Editorial Board, headed by the Editors-in-Chief, Greg Barsh (HudsonAlpha Institute of Biotechnology, and Stanford University School of Medicine) and Greg Copenhaver (The University of North Carolina at Chapel Hill). Articles published in PLOS Genetics are archived in PubMed Central and cited in PubMed.
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