title: In vitro interaction between coxsackievirus B3 VP1 protein and human pleckstrin homology domain retinal protein (PHR1).
abstract: Coxsackievirus B3 (CVB3) infection causes central nervous system diseases including aseptic meningitis and encephalitis. To understand the mechanism of this virus, a yeast two-hybrid system was used to screen cellular proteins from a human heart cDNA library. The results revealed that the human Pleckstrin Homology Domain Retinal protein (PHR1), a PH domain-containing protein with low expression in the heart and high expression in the brain, interacts with CVB3 VP1, a major structural protein of CVB3. Yeast mating assays and in vitro coimmunoprecipitation verified the interaction between CVB3 VP1 and PHR1. An α-galactosidase assay indicated that of α-galactosidase activity was higher in positive clones than in controls suggesting a strong interaction. Furthermore, assay of deletion mutants defined the minimal region of PHR1 required for its interaction with VP1 as amino acids 95-172 and two regions of VP1 required for its interaction with PHR1 as amino acids 729-767 and 811-859. The results revealed multiple binding sites between PHR1 and CVB3 VP1 and suggested that the strong interaction between these two proteins might play an important role in central nervous system disease in the human brain.
authors: Basic Medical College of Nanchang University, Nanchang, 330006, People's Republic of China.
authors: Ying
authors: Y
authors: Zhou
authors: Basic Medical College of Nanchang University, Nanchang, 330006, People's Republic of China.
authors: Zhiqin
authors: Z
authors: Zhang
authors: The People's Hospital in Jiangxi Province, Nanchang, 330006, People's Republic of China.
authors: Hongluan
authors: H
authors: Wang
authors: Basic Medical College of Nanchang University, Nanchang, 330006, People's Republic of China.
authors: Yanhua
authors: Y
authors: Xia
authors: Basic Medical College of Nanchang University, Nanchang, 330006, People's Republic of China.
authors: Xiuzhen
authors: X
authors: Li
authors: Basic Medical College of Nanchang University, Nanchang, 330006, People's Republic of China.
authors: Yan
authors: Y
authors: Yan
authors: Basic Medical College of Nanchang University, Nanchang, 330006, People's Republic of China.
authors: Weiwen
authors: W
authors: Zou
authors: Department of Clinical Laboratory, The First Affiliated Hospital of Nanchang University, Nanchang University, Nanchang, 330006, People's Republic of China.
authors: Lingbing
authors: L
authors: Zeng
authors: Basic Medical College of Nanchang University, Nanchang, 330006, People's Republic of China. xthuang@ncu.edu.cn.
authors: Xiaotian
authors: X
authors: Huang
mesh_terms: D004252:DNA Mutational Analysis; D029822:Enterovirus B, Human / Q000502:physiology*; D015723:Gene Library; D054884:Host-Pathogen Interactions; D006801:Humans; D047468:Immunoprecipitation; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D025941:Protein Interaction Mapping; D020798:Two-Hybrid System Techniques; D015678:Viral Structural Proteins / Q000378:metabolism*
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C417163:PLEKHB1 protein, human; D015678:Viral Structural Proteins
keywords: Coxsackievirus B3 VP1; Interaction; Pleckstrin homology domain retinal protein (PHR1)