Epstein-Barr virus nuclear antigen 1 is a DNA binding protein with strong RNA-binding activity
Journal
Journal of General Virology
Journal Volume
85
Journal Issue
10
Pages
2755-2765
Date Issued
2004
Author(s)
Abstract
Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA-1) plays key roles in both the regulation of gene expression and the replication of the EBV genome in latently infected cells. To characterize the RNA-binding activity of EBNA-1, it was demonstrated that EBNA-1 binds efficiently to RNA homopolymers that are composed of poly(G) and weakly to those composed of poly(U). All three RGG boxes of EBNA-1 contributed additively to poly(G)-binding activity and could mediate RNA binding when attached to a heterologous protein in an RNA gel mobility-shift assay. In vitro-transcribed EBV and non-EBV RNA probes revealed that EBNA-1 bound to most RNAs examined and the affinity increased as the content of G and U increased, as demonstrated in competition assays. Among these probes, the 5′ non-coding region (NCR) (nt 131-278) of hepatitis C virus RNA appeared to be the strongest competitor for EBNA-1 binding to the EBV-encoded small nuclear RNA 1 (EBER1) probe, whereas a mutant 5′ NCR RNA with partially disrupted secondary structure was a weak competitor. Furthermore, the interaction of endogenous EBNA-1 and EBER1 in EBV-infected cells was demonstrated by a ribonucleoprotein immunoprecipitation assay. These results revealed that EBNA-1 is a DNA-binding protein with strong binding activity to a relatively broad spectrum of RNA and suggested an additional biological impact of EBNA-1 through its ability to bind to RNA. ? 2004 SGM.
SDGs
Other Subjects
cell nucleus antigen; DNA binding protein; polyguanylic acid; polyuridylic acid; ribonucleoprotein; RNA; small nuclear RNA; untranslated RNA; virus RNA; DNA binding protein; EBV encoded nuclear antigen 1; EBV-encoded nuclear antigen 1; Epstein Barr virus antigen; Epstein Barr virus encoded RNA 1; Epstein-Barr virus encoded RNA 1; recombinant protein; RNA; article; binding affinity; binding competition; Epstein Barr virus; gel mobility shift assay; gene expression regulation; gene replication; Hepatitis C virus; immunoprecipitation; in vitro study; molecular interaction; nonhuman; nucleotide sequence; priority journal; protein binding; RNA binding; RNA probe; RNA structure; virus genome; virus infection; chemistry; genetics; metabolism; serodiagnosis; Hepatitis C virus; Human herpesvirus 4; DNA-Binding Proteins; Epstein-Barr Virus Nuclear Antigens; Hepacivirus; Precipitin Tests; Recombinant Proteins; RNA; RNA, Viral
Type
journal article