Epstein-Barr virus nuclear antigen-1 is highly colocalized with interphase chromatin and its newly replicated regions in particular.
Epstein-Barr virus (EBV) nuclear antigen-1 (EBNA-1), which binds to both the EBV origin of replication (oriP) and metaphase chromosomes, is essential for the replication/retention and segregation/partition of oriP-containing plasmids. Here the chromosomal localization of EBNA-1 fused to green fluorescent protein (GFP-EBNA-1) is examined by confocal microscopy combined with a 'premature chromosome condensation' (PCC) procedure. Analyses show that GFP-EBNA-1 expressed in living cells that lack oriP plasmids is associated with cellular chromatin that has been condensed rapidly by the PCC procedure into identifiable forms that are unique to each phase of interphase as well as metaphase chromosomes. Studies of cellular chromosomal DNAs labelled with BrdU or Cy3-dUTP indicate that GFP-EBNA-1 colocalizes highly with the labelled, newly replicated regions of interphase chromatin in cells. These results suggest that EBNA-1 is associated not only with cellular metaphase chromosomes but also with condensing chromatin/chromosomes and probably with interphase chromatin, especially with its newly replicated regions.
Ito
Sayuri
S
Department of Virology I1 and Division of Genetic Resources2, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku, Tokyo, 162-8640, Japan.
Gotoh
Eisuke
E
Department of Virology I1 and Division of Genetic Resources2, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku, Tokyo, 162-8640, Japan.
Ozawa
Shigeru
S
Department of Microbiology, Yamanashi Institute of Health, Fujimi 1-7-31, Kofu City, Yamanashi Prefecture, Japan3.|Department of Virology I1 and Division of Genetic Resources2, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku, Tokyo, 162-8640, Japan.
Yanagi
Kazuo
K
Department of Virology I1 and Division of Genetic Resources2, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku, Tokyo, 162-8640, Japan.
D000818:Animals; D016466:CHO Cells; D002232:Carbocyanines; D002843:Chromatin / Q000378:metabolism*; D006224:Cricetinae; D004279:DNA, Viral / Q000378:metabolism*; D003856:Deoxyuracil Nucleotides; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics / Q000378:metabolism*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000378:metabolism*; D007399:Interphase; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D018741:Replication Origin; D014779:Virus Replication / Q000502:physiology*
C088941:3'-deoxy-5-(cyanine dye 3)uridine 5'-trisphosphate; D002232:Carbocyanines; D002843:Chromatin; D004279:DNA, Viral; D003856:Deoxyuracil Nucleotides; D019309:Epstein-Barr Virus Nuclear Antigens; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; C417029:EBV-encoded nuclear antigen 1