Global regulatory roles of the cAMP/PKA pathway revealed by phenotypic, transcriptomic and phosphoproteomic analyses in a null mutant of the PKA catalytic subunit in Candida albicans
Global regulatory roles of the cAMP/PKA pathway revealed by phenotypic, transcriptomic and phosphoproteomic analyses in a null mutant of the PKA catalytic subunit in Candida albicans
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通过表型、转录组和磷酸蛋白质组分析揭示白色念珠菌 PKA 催化亚基无效突变体中 cAMP/PKA 通路的全局调节作用
DOI：
10.1111/mmi.13681
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发表时间：
2017-07-01
影响因子：
3.6
通讯作者：
Huang, Guanghua
中科院分区：
文献类型：
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作者：
Cao, Chengjun;Wu, Mei;Huang, Guanghua
保守的cAMP依赖蛋白激酶(PKA)在人类真菌病原菌白色念珠菌的形态转变和毒力调节中起着关键作用。长期以来，人们一直认为PKA催化亚基对这种真菌的细胞活力是必不可少的。矛盾的是，白念珠菌产生cAMP所需的单一腺苷环化酶编码基因Cyr1并不是细胞生长所必需的。在这里，成功地产生了TPK1和TPK2(tpk2/tpk2，tpk1/tpk1，t2t1)的双突变体，它们编码两种形式的PKA催化亚基，表明该亚基对细胞活力不是必需的。PKA催化亚基的失活阻止了丝状化，显著地减弱了从白色到不透明的转换，但促进了有性交配。比较转录学分析表明，t2t1和cyr1/cyr1突变体具有相似的全球基因表达谱。与WT株相比，t2t1和cyr1/cyr1突变株的总转录活性和代谢均显著降低。结合磷蛋白组学和生物信息学分析，我们确定了181个潜在的PKA磷酸化靶点，代表了148个独特的蛋白质，参与了广泛的生物过程。这项研究为白念珠菌cAMP/PKA途径的全球调控特征提供了新的见解。
The conserved cAMP-dependent protein kinase (PKA) plays critical roles in the regulation of morphological transitions and virulence in the human fungal pathogen Candida albicans. It has long been thought that the PKA catalytic subunit is essential for cell viability in this fungus. Paradoxically, the single adenylyl cyclase-encoding gene, CYR1, which is required for the production of cAMP in C. albicans, is not essential for cell growth. Here, a double mutant of TPK1 and TPK2 (tpk2/tpk2 tpk1/tpk1, t2t1), which encode two isoforms of the PKA catalytic subunit was successfully generated, suggesting that this subunit is not essential for cell viability. Inactivation of the PKA catalytic subunit blocked filamentation and dramatically attenuated white-to-opaque switching, but promoted sexual mating. Comparative transcriptomic analyses demonstrated that the t2t1 and cyr1/cyr1 mutants exhibited similar global gene expression profiles. Compared with the WT strain, the general transcriptional activity and metabolism were significantly decreased in both the t2t1 and cyr1/cyr1 mutants. Using combined phosphoproteomic and bioinformatic analyses, we identified 181 potential PKA phosphorylation targets, which represent 148 unique proteins involved in a wide spectrum of biological processes. The study sheds new insights into the global regulatory features of the cAMP/PKA pathway in C. albicans.