Identification and characterization of PhbF: a DNA binding protein with regulatory role in the PHB metabolism of Herbaspirillum seropedicae SmR1.
Identification and characterization of PhbF: a DNA binding protein with regulatory role in the PHB metabolism of Herbaspirillum seropedicae SmR1.
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DOI：
10.1186/1471-2180-11-230
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发表时间：
2011-10-14
期刊：
影响因子：
4.2
通讯作者：
Steffens MB
中科院分区：
文献类型：
--
作者：
Kadowaki MA;Müller-Santos M;Rego FG;Souza EM;Yates MG;Monteiro RA;Pedrosa FO;Chubatsu LS;Steffens MB
Herbaspirillum seropedicae SmR1是一种重要农作物的固氮内生菌。它产生聚羟基丁酸酯（PHB），其作为颗粒储存在细胞内。然而，在这种生物体中，PHB代谢和调节控制尚未得到很好的研究。在这项工作中，我们描述了来自H.在大肠杆菌中表达后，纯化并鉴定了血清型SmR1。杆菌纯化的PhbF蛋白能够与H.血清型SmR1。计算机模拟分析表明可能的DNA结合序列，其在使用纯化的PhbF的DNA足迹分析中显示出受到保护。使用lacZ融合的分析表明，PhbF可以作为一个阻遏蛋白控制的PHB代谢相关基因的表达。结果表明，H. Seropedicae SmR1 PhbF通过作为转录抑制因子调节phb相关基因的表达。了解这种植物共生细菌的PHB代谢，有助于了解植物定殖过程以及生物体的抗性和在植物中的存活。
Herbaspirillum seropedicae SmR1 is a nitrogen fixing endophyte associated with important agricultural crops. It produces polyhydroxybutyrate (PHB) which is stored intracellularly as granules. However, PHB metabolism and regulatory control is not yet well studied in this organism. In this work we describe the characterization of the PhbF protein from H. seropedicae SmR1 which was purified and characterized after expression in E. coli. The purified PhbF protein was able to bind to eleven putative promoters of genes involved in PHB metabolism in H. seropedicae SmR1. In silico analyses indicated a probable DNA-binding sequence which was shown to be protected in DNA footprinting assays using purified PhbF. Analyses using lacZ fusions showed that PhbF can act as a repressor protein controlling the expression of PHB metabolism-related genes. Our results indicate that H. seropedicae SmR1 PhbF regulates expression of phb-related genes by acting as a transcriptional repressor. The knowledge of the PHB metabolism of this plant-associated bacterium may contribute to the understanding of the plant-colonizing process and the organism's resistance and survival in planta.
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