Analysis of the Candida albicans Phosphoproteome.

Eukaryotic Cell Pub Date : 2015-05-01 Epub Date: 2015-03-06 DOI:10.1128/EC.00011-15
S D Willger, Z Liu, R A Olarte, M E Adamo, J E Stajich, L C Myers, A N Kettenbach, D A Hogan
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引用次数: 38

Abstract

Candida albicans is an important human fungal pathogen in both immunocompetent and immunocompromised individuals. C. albicans regulation has been studied in many contexts, including morphological transitions, mating competence, biofilm formation, stress resistance, and cell wall synthesis. Analysis of kinase- and phosphatase-deficient mutants has made it clear that protein phosphorylation plays an important role in the regulation of these pathways. In this study, to further our understanding of phosphorylation in C. albicans regulation, we performed a deep analysis of the phosphoproteome in C. albicans. We identified 19,590 unique peptides that corresponded to 15,906 unique phosphosites on 2,896 proteins. The ratios of serine, threonine, and tyrosine phosphosites were 80.01%, 18.11%, and 1.81%, respectively. The majority of proteins (2,111) contained at least two detected phosphorylation sites. Consistent with findings in other fungi, cytoskeletal proteins were among the most highly phosphorylated proteins, and there were differences in Gene Ontology (GO) terms for proteins with serine and threonine versus tyrosine phosphorylation sites. This large-scale analysis identified phosphosites in protein components of Mediator, an important transcriptional coregulatory protein complex. A targeted analysis of the phosphosites in Mediator complex proteins confirmed the large-scale studies, and further in vitro assays identified a subset of these phosphorylations that were catalyzed by Cdk8 (Ssn3), a kinase within the Mediator complex. These data represent the deepest single analysis of a fungal phosphoproteome and lay the groundwork for future analyses of the C. albicans phosphoproteome and specific phosphoproteins.

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白色念珠菌磷蛋白组分析。
白色念珠菌是一种重要的人类真菌病原体在免疫正常和免疫功能低下的个体。白色念珠菌的调控在许多方面都得到了研究,包括形态转变、交配能力、生物膜形成、抗逆性和细胞壁合成。对激酶和磷酸酶缺陷突变体的分析表明,蛋白质磷酸化在这些途径的调控中起着重要作用。在本研究中,为了进一步了解白色念珠菌的磷酸化调控,我们对白色念珠菌的磷酸化蛋白组进行了深入分析。我们鉴定了19,590个独特的肽,对应于2,896个蛋白质上的15,906个独特的磷酸基。丝氨酸、苏氨酸和酪氨酸磷酸位点的比例分别为80.01%、18.11%和1.81%。大多数蛋白(2,111)含有至少两个检测到的磷酸化位点。与其他真菌的发现一致,细胞骨架蛋白是磷酸化程度最高的蛋白质之一,并且在基因本体(GO)术语中,丝氨酸和苏氨酸与酪氨酸磷酸化位点的蛋白质存在差异。这种大规模的分析鉴定了Mediator蛋白组分中的磷位点,这是一种重要的转录协同调节蛋白复合物。对Mediator复合体蛋白中磷酸化位点的靶向分析证实了大规模的研究,进一步的体外实验确定了这些磷酸化的一个子集,这些磷酸化是由Cdk8 (Ssn3)催化的,这是Mediator复合体中的一种激酶。这些数据代表了真菌磷蛋白组的最深入的单一分析,并为今后白念珠菌磷蛋白组和特异性磷蛋白的分析奠定了基础。
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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来源期刊
Eukaryotic Cell
Eukaryotic Cell 生物-微生物学
自引率
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审稿时长
1 months
期刊介绍: Eukaryotic Cell (EC) focuses on eukaryotic microbiology and presents reports of basic research on simple eukaryotic microorganisms, such as yeasts, fungi, algae, protozoa, and social amoebae. The journal also covers viruses of these organisms and their organelles and their interactions with other living systems, where the focus is on the eukaryotic cell. Topics include: - Basic biology - Molecular and cellular biology - Mechanisms, and control, of developmental pathways - Structure and form inherent in basic biological processes - Cellular architecture - Metabolic physiology - Comparative genomics, biochemistry, and evolution - Population dynamics - Ecology
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