title: Nucleolin is important for Epstein-Barr virus nuclear antigen 1-mediated episome binding, maintenance, and transcription.
abstract: Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA1) is essential for EBV episome maintenance, replication, and transcription. These effects are mediated by EBNA1 binding to cognate oriP DNA, which comprise 20 imperfect copies of a 30-bp dyad symmetry enhancer and an origin for DNA replication. To identify cell proteins essential for these EBNA1 functions, EBNA1 associated cell proteins were immune precipitated and analyzed by liquid chromatography-tandem mass spectrometry. Nucleolin (NCL) was identified to be EBNA1 associated. EBNA1's N-terminal 100 aa and NCL's RNA-binding domains were critical for EBNA1/NCL interaction. Lentivirus shRNA-mediated NCL depletion substantially reduced EBNA1 recruitment to oriP DNA, EBNA1-dependent transcription of an EBV oriP luciferase reporter, and EBV genome maintenance in lymphoblastoid cell lines. NCL RNA-binding domain K429 was critical for ATP and EBNA1 binding. NCL overexpression increased EBNA1 binding to oriP and transcription, whereas NCL K429A was deficient. Moreover, NCL silencing impaired lymphoblastoid cell line growth. These experiments reveal a surprisingly critical role for NCL K429 in EBNA1 episome maintenance and transcription, which may be a target for therapeutic intervention.
authors.affiliation: Department of Life Sciences, Tzu-Chi University, Hualien 97004, Taiwan.
authors.forename: Ya-Lin
authors.initials: YL
authors.lastname: Chen
authors.forename: Cheng-Der
authors.initials: CD
authors.lastname: Liu
authors.forename: Chi-Ping
authors.initials: CP
authors.lastname: Cheng
authors.forename: Bo
authors.initials: B
authors.lastname: Zhao
authors.forename: Hao-Jen
authors.initials: HJ
authors.lastname: Hsu
authors.forename: Chih-Long
authors.initials: CL
authors.lastname: Shen
authors.forename: Shu-Jun
authors.initials: SJ
authors.lastname: Chiu
authors.forename: Elliott
authors.initials: E
authors.lastname: Kieff
authors.forename: Chih-wen
authors.initials: CW
authors.lastname: Peng
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D001665:Binding Sites; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002853:Chromatography, Liquid; D004261:DNA Replication; D000939:Epitopes / Q000737:chemistry; D019309:Epstein-Barr Virus Nuclear Antigens / Q000737:chemistry* / Q000502:physiology*; D005786:Gene Expression Regulation; D020868:Gene Silencing; D016678:Genome; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D008156:Luciferases / Q000378:metabolism; D013058:Mass Spectrometry; D018613:Microscopy, Confocal; D010750:Phosphoproteins / Q000737:chemistry* / Q000502:physiology*; D010957:Plasmids / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000502:physiology*; D018741:Replication Origin; D014158:Transcription, Genetic; D014779:Virus Replication; D000097796:Nucleolin
chemical_list: D000255:Adenosine Triphosphate; D000939:Epitopes; D019309:Epstein-Barr Virus Nuclear Antigens; D049452:Green Fluorescent Proteins; D008156:Luciferases; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D000097796:Nucleolin; C417029:EBV-encoded nuclear antigen 1
keywords: chromatin; lymphoma; nasopharyngeal carcinoma; oncogenic herpesvirus