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Serial Analysis of Gene Expression Reveals Conserved Links between Protein Kinase A Ribosome Biogenesis and Phosphate Metabolism in Ustilago maydis

机译:基因表达的系列分析揭示了美til鱼中蛋白激酶A核糖体生物发生和磷酸盐代谢之间的保守联系。

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摘要

The switch from budding to filamentous growth is a key aspect of invasive growth and virulence for the fungal phytopathogen Ustilago maydis. The cyclic AMP (cAMP) signaling pathway regulates dimorphism in U. maydis, as demonstrated by the phenotypes of mutants with defects in protein kinase A (PKA). Specifically, a mutant lacking the regulatory subunit of PKA encoded by the ubc1 gene displays a multiple-budded phenotype and fails to incite disease symptoms, although proliferation does occur in the plant host. A mutant with a defect in a catalytic subunit of PKA, encoded by adr1, has a constitutively filamentous phenotype and is nonpathogenic. We employed serial analysis of gene expression to examine the transcriptomes of a wild-type strain and the ubc1 and adr1 mutants to further define the role of PKA in U. maydis. The mutants displayed changes in the transcript levels for genes encoding ribosomal proteins, genes regulated by the b mating-type proteins, and genes for metabolic functions. Importantly, the ubc1 mutant displayed elevated transcript levels for genes involved in phosphate acquisition and storage, thus revealing a connection between cAMP and phosphate metabolism. Further experimentation indicated a phosphate storage defect and elevated acid phosphatase activity for the ubc1 mutant. Elevated phosphate levels in culture media also enhanced the filamentous growth of wild-type cells in response to lipids, a finding consistent with PKA regulation of morphogenesis in U. maydis. Overall, these findings extend our understanding of cAMP signaling in U. maydis and reveal a link between phosphate metabolism and morphogenesis.
机译:从发芽到丝状生长的转变是真菌植物病原菌Ustilago maydis侵袭性生长和毒力的关键方面。循环AMP(cAMP)信号通路调节美人鱼中的二态性,如蛋白激酶A(PKA)中存在缺陷的突变体的表型所证明。具体而言,尽管在植物宿主中确实发生了增殖,但缺乏由ubc1基因编码的PKA调节亚基的突变体表现出多重预算表型,并且没有引起疾病症状。由adr1编码的PKA催化亚基缺陷的突变体具有组成型丝状表型,并且是非致病性的。我们使用基因表达的系列分析来检查野生型菌株和ubc1和adr1突变体的转录组,以进一步定义PKA在美人鱼中的作用。突变体显示出编码核糖体蛋白的基因,受交配型蛋白调控的基因以及代谢功能基因的转录水平变化。重要的是,ubc1突变体显示参与磷酸盐获取和储存的基因的转录水平升高,从而揭示了cAMP与磷酸盐代谢之间的联系。进一步的实验表明ubc1突变体的磷酸盐存储缺陷和酸性磷酸酶活性升高。培养基中磷酸盐水平的升高还增强了响应脂质对野生型细胞丝状生长的影响,这一发现与梅花。鱼形态发生的PKA调节一致。总体而言,这些发现扩展了我们对美人鱼中cAMP信号传导的理解,并揭示了磷酸盐代谢与形态发生之间的联系。

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