title: Overexpression of microRNA gga-miR-21 in chicken fibroblasts suppresses replication of infectious bursal disease virus through inhibiting VP1 translation.
abstract: Previously we have identified a series of cellular miRNA molecules up- or down-regulated in infectious bursal disease virus (IBDV) infected chicken embryo fibroblasts and Bursa of Fabricius with gene microarray analysis. Here we studied in detail a relatively well studied miRNA, gga-miR-21, for better understanding miRNAs involvement in IBDV-host interactions. Chicken pri-gga-miRNA-21 and a control miRNA Caenorhabditis elegans pri-cel-lin-4 gene were cloned into a lentiviral vector, respectively. The resulting recombinant lentiviruses were used to infect chicken fibroblast cell line DF-1, and two stable cell lines, DF-miR-21 (overexpressing gga-miR-21) and DF-lin-4 (overexpressing cel-lin-4), were selected. Replication of IBDV in DF-miR-21, DF-lin-4 and DF-1 cells were compared and molecular mechanism of IBDV replication alteration was explored using bioinformatics, reporter gene system, qRT-PCR and Western blot analysis. IBDV replication was markedly lower in DF-miR-21 than in DF-lin-4 or DF-1 cells. A gga-miR-21 target sequence was identified within IBDV VP1 gene (1713-1734bp). Fusion of a 520nt long partial IBDV VP1 gene containing the target with a luciferase gene resulted in significantly lower transient luciferase activity in DF-miR-21 cells as compared to that in DF-lin-4 or DF-1 cell. Following IBDV infection of the cell lines, VP1 protein level in DF-miR-21 cells was dramatically lower than that in DF-lin-4 or DF-1 cells but VP1 mRNA level was not different. The finding indicated that gga-miR-21 could suppress IBDV replication through down regulating IBDV VP1 expression at translational level.
authors: Key Laboratory of Veterinary Biological Engineering and Technology, Ministry of Agriculture, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China. Electronic address: wangys63@126.com.
authors: Yong-Shan
authors: YS
authors: Wang
authors: Wei
authors: W
authors: Ouyang
authors: Qun-Xing
authors: QX
authors: Pan
authors: Xiao-Li
authors: XL
authors: Wang
authors: Xing-Xia
authors: XX
authors: Xia
authors: Zhen-Wei
authors: ZW
authors: Bi
authors: Yong-Qiang
authors: YQ
authors: Wang
authors: Xiao-Mei
authors: XM
authors: Wang
mesh_terms: D000818:Animals; D018175:Birnaviridae Infections / Q000235:genetics / Q000378:metabolism / Q000662:veterinary* / Q000821:virology; D002642:Chick Embryo; D002645:Chickens; D015536:Down-Regulation; D005347:Fibroblasts / Q000378:metabolism / Q000821:virology*; D007243:Infectious bursal disease virus / Q000235:genetics / Q000502:physiology*; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D011201:Poultry Diseases / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D014176:Protein Biosynthesis; D015678:Viral Structural Proteins / Q000235:genetics* / Q000378:metabolism; D014779:Virus Replication
chemical_list: D035683:MicroRNAs; C107669:VP1 protein, infectious bursal disease virus; D015678:Viral Structural Proteins
keywords: Infectious bursal disease virus (IBDV); Virus replication; gga-miR-21; microRNA (miRNA)