title: Identification of NS1 domains of avian H5N1 influenza virus which influence the interaction with the NOLC1 protein.
abstract: Non-structural protein 1 (NS1) is an important virulence factor encoded by influenza A virus. NS1 can interact with a variety of host cell proteins to interfere with the host innate immune response and to promote effective viral replication. Our previous work has shown that only the effector domain of NS1 (amino acid residues 74-230/237) is sufficient to interact with nucleolar and coiled-body phosphoprotein 1 (NOLC1). To investigate the exact region of NS1 that interacts with NOLC1, we used only the effector domain of NS1 and constructed various mutants having different deletions, and then tested their ability to interact with NOLC1 via pull-down assay. Only the mutant containing amino acid residues 104-200 showed positive interaction with NOLC1. To further determine the key amino acids of the NS1 effector domain which are crucial for interaction with NOLC1, several mutants containing a single amino acid substitution were made and their interaction with NOLC1 was tested. Only the mutant D120A or R195A showed reduced binding with NOLC1, suggesting that D120 and R195 were crucial to the binding of NS1 to NOLC1. This study lays the foundation for further research aiming at furthering our understanding of the interaction between NS1 and host cells.
authors: Key Laboratory of Animal Resource and Epidemic Disease Prevention of Liaoning Province, College of Life Science of Liaoning University, Shenyang, 110036, People's Republic of China.
authors: Chun-Yu
authors: CY
authors: Zhu
authors: Fang-Liang
authors: FL
authors: Zheng
authors: Xiao-Shuang
authors: XS
authors: She
authors: Dan
authors: D
authors: Zhao
authors: Ying
authors: Y
authors: Gu
authors: Yan-Ting
authors: YT
authors: Duan
authors: Alan K
authors: AK
authors: Chang
authors: Hong-Sheng
authors: HS
authors: Liu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007251:Influenza, Human / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014774:Virulence
chemical_list: C085820:INS1 protein, influenza virus; C075564:NOLC1 protein, human; D009687:Nuclear Proteins; D010750:Phosphoproteins; D017361:Viral Nonstructural Proteins
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