Mutational analysis of the Epstein-Barr virus nuclear antigen 2 by far-Western blotting and DNA-binding studies.
We have previously shown by far-Western blotting that the Epstein-Barr virus nuclear antigen 2 (EBNA-2) both binds to a cellular protein of 130 kDa and histone H1, with the complex between EBNA-2 and p130 being tighter than between EBNA-2 and histone H1. Here we demonstrate that the N terminus of EBNA-2, which was previously shown to be necessary for transformation of B lymphocytes by EBNA-2, is essential for binding to p130. We further show data indicating that the binding of EBNA-2 to histone H1 appears not to be mediated exclusively via the basic Arg-Gly rich region in the C-terminal part of EBNA-2. With a MAb directed against the Trp-Trp322-Pro (WWP) motif of EBNA-2, which is known to be essential for the interaction of EBNA-2 with the cellular factor RBPJkappa/CBF1, we could inhibit the DNA binding of EBNA-2, providing further evidence that this region of EBNA-2 forms direct contact with RBPJkappa/CBF1.
Sauder
C
C
Institut für medizinische Mikrobiologie und Hygiene der Universitätskliniken des Saarlandes.
Götzinger
N
N
Schubach
W H
WH
Horvath
G C
GC
Kremmer
E
E
Krebs
A
A
König
S
S
Zimber Strobl
U
U
Mueller Lantzsch
N
N
Grässer
F A
FA
D000595:Amino Acid Sequence; D000956:Antigens, Viral / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D004247:DNA / Q000378:metabolism*; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D019309:Epstein-Barr Virus Nuclear Antigens; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006657:Histones / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013329:Structure-Activity Relationship
D000956:Antigens, Viral; D004268:DNA-Binding Proteins; D019309:Epstein-Barr Virus Nuclear Antigens; D006657:Histones; D004247:DNA