Boyce Kylie J, Kretschmer Matthias, Kronstad James W
The Michael Smith Laboratories, Department of Microbiology and Immunology, and Faculty of Land and Food Systems, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4.
Eukaryot Cell. 2006 Aug;5(8):1399-409. doi: 10.1128/EC.00131-06.
The maize pathogen Ustilago maydis switches from budding to filamentous, dikaryotic growth in response to environmental signals including nutrient status, growth in the host, and the presence of mating pheromones. The filamentous dikaryon is capable of proliferating within host tissue to cause disease symptoms including tumors. The transition from yeast cells to hyphal filaments is regulated by a mitogen-activated protein kinase cascade and a cyclic-AMP-protein kinase A (PKA) pathway. Serial analysis of gene expression with PKA mutants identified orthologs of components of the PHO phosphate acquisition pathway as transcriptional targets of the PKA pathway, and these included genes for Pho84, an acid phosphatase, and the vacuolar transport chaperones Vtc1 and Vtc4. In Saccharomyces cerevisiae, Vtc4p is required during the fusion of inorganic-phosphate-containing vesicles to the vacuolar membrane and the consequent accumulation of phosphate stored as polyphosphate (polyP) in the vacuole. We found that deletion of vtc4 in U. maydis also reduced polyP stored in vacuoles. Intriguingly, Deltavtc4 mutants possessed a filamentous cellular morphology, in contrast to the budding, yeast-like growth of the wild-type parent. The Deltavtc4 mutants also displayed decreased symptom development and reduced proliferation in planta. The interaction with PKA signaling was further investigated by the generation of Deltavtc4 ubc1 double mutants. Deletion of vtc4 completely suppressed the multiple-budded phenotype of a Deltaubc1 mutant, indicating that polyP stores are essential for this PKA-induced trait. Overall, this study reveals a novel role for PKA-regulated polyP accumulation in the control of fungal morphogenesis and virulence.
玉米病原体玉米黑粉菌(Ustilago maydis)会根据包括营养状况、在宿主中的生长情况以及交配信息素的存在等环境信号,从出芽生长转变为丝状双核生长。丝状双核体能够在宿主组织内增殖,引发包括肿瘤在内的疾病症状。从酵母细胞到菌丝的转变受丝裂原活化蛋白激酶级联反应和环磷酸腺苷 - 蛋白激酶A(PKA)途径调控。对PKA突变体进行基因表达序列分析,确定了PHO磷酸盐获取途径成分的直系同源物为PKA途径的转录靶点,其中包括酸性磷酸酶Pho84以及液泡运输伴侣蛋白Vtc1和Vtc4的基因。在酿酒酵母(Saccharomyces cerevisiae)中,含无机磷酸盐的囊泡与液泡膜融合以及随后液泡中以多聚磷酸盐(polyP)形式储存的磷酸盐积累过程中需要Vtc4p。我们发现,在玉米黑粉菌中缺失vtc4也会减少液泡中储存的多聚磷酸盐。有趣的是,与野生型亲本的出芽酵母样生长不同,Δvtc4突变体具有丝状细胞形态。Δvtc4突变体在植物中的症状发展也有所减少,增殖能力降低。通过构建Δvtc4 ubc1双突变体进一步研究了其与PKA信号传导的相互作用。缺失vtc4完全抑制了Δubc1突变体的多芽表型,表明多聚磷酸盐储存对于这种PKA诱导的性状至关重要。总体而言,本研究揭示了PKA调节的多聚磷酸盐积累在控制真菌形态发生和毒力方面的新作用。