Müller Karel, Doubnerová Veronika, Synková Helena, Cerovská Noemi, Ryslavá Helena
Department of Biochemistry, Faculty of Natural Science, Charles University, Hlavova 2030, CZ-128 00 Prague 2, Czech Republic.
Biol Chem. 2009 Mar;390(3):245-51. doi: 10.1515/BC.2009.029.
The effect of viral infection on the regulation of phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) in Nicotiana tabacum L. leaves was studied. PEPC activity was 3 times higher in infected plant leaves compared to healthy plants. Activity of plant PEPC can be regulated, e.g., by de novo synthesis or reversible phosphorylation. The reason for the increase of PEPC activity as a consequence of PVY(NTN) infection was studied. The amount of PEPC determined by Western blot analysis or by relative estimation of PEPC mRNA by real-time PCR did not differ in control and PVY(NTN)-infected plants. Changes in posttranslational modification of PEPC by phosphorylation were evaluated by comparing activity of the native and the dephosphorylated enzyme. The infected plants were characterized by a higher decrease of the enzyme activity after its dephosphorylation, which indicated a higher phosphorylation level. Immunochemical detection of phosphoproteins by Western blot analysis showed a more intensive band corresponding to PEPC from the infected material. This strengthens the hypothesis of an infection-related phosphorylation, which could be part of the plant's response to pathogen attack. The physiological implications of the increase in PEPC activity during PVY(NTN) infection are discussed.
研究了病毒感染对烟草叶片中磷酸烯醇式丙酮酸羧化酶(PEPC,EC 4.1.1.31)调控的影响。与健康植株相比,感染植株叶片中的PEPC活性高3倍。植物PEPC的活性可通过例如从头合成或可逆磷酸化来调控。研究了PVY(NTN)感染导致PEPC活性增加的原因。通过蛋白质免疫印迹分析测定的PEPC量或通过实时PCR对PEPC mRNA进行相对定量,在对照植株和PVY(NTN)感染植株中并无差异。通过比较天然酶和去磷酸化酶的活性,评估了磷酸化对PEPC翻译后修饰的影响。感染植株的特点是去磷酸化后酶活性的降低幅度更大,这表明磷酸化水平更高。通过蛋白质免疫印迹分析对磷蛋白进行免疫化学检测,结果显示来自感染材料的与PEPC对应的条带更强。这强化了与感染相关的磷酸化假说,这可能是植物对病原体攻击反应的一部分。文中讨论了PVY(NTN)感染期间PEPC活性增加的生理意义。