Larraya Luis M, Boyce Kylie J, So Austin, Steen Barbara R, Jones Steven, Marra Marco, Kronstad James W
Michael Smith Laboratories, 2185 East Mall, University of British Columbia, Vancouver, BC V6T 1Z4, Canada.
Eukaryot Cell. 2005 Dec;4(12):2029-43. doi: 10.1128/EC.4.12.2029-2043.2005.
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.
从出芽生长转变为丝状生长是真菌植物病原体玉米黑粉菌侵袭性生长和毒力的一个关键方面。环磷酸腺苷(cAMP)信号通路调节玉米黑粉菌的二态性,这已通过蛋白激酶A(PKA)缺陷突变体的表型得到证明。具体而言,一个缺乏由ubc1基因编码的PKA调节亚基的突变体表现出多芽表型,并且不能引发疾病症状,尽管在植物宿主中确实会发生增殖。一个由adr1编码的PKA催化亚基存在缺陷的突变体具有组成型丝状表型,并且是非致病性的。我们采用基因表达序列分析来检测野生型菌株以及ubc1和adr1突变体的转录组,以进一步确定PKA在玉米黑粉菌中的作用。这些突变体在编码核糖体蛋白的基因、由b交配型蛋白调控的基因以及代谢功能相关基因的转录水平上出现了变化。重要的是,ubc1突变体中参与磷酸盐获取和储存的基因转录水平升高,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突变体存在磷酸盐储存缺陷且酸性磷酸酶活性升高。培养基中升高的磷酸盐水平也增强了野生型细胞对脂质的丝状生长反应,这一发现与PKA对玉米黑粉菌形态发生的调节一致。总体而言,这些发现扩展了我们对玉米黑粉菌中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。