Characterization of DP103, a novel DEAD box protein that binds to the Epstein-Barr virus nuclear proteins EBNA2 and EBNA3C.
The Epstein-Barr virus-encoded nuclear antigens EBNA2 and EBNA3C both interact with the cellular transcription factor RBP-Jkappa and modulate the expression of several shared target genes, suggesting a tight cooperation in latently infected cells. In a survey for additional cellular factors that bind to EBNA2 as well as EBNA3C, we have isolated and characterized DP103, a novel human member of the DEAD box family of putative ATP-dependent RNA helicases. The interaction with DP103 is mediated by amino acids (aa) 121-213 of EBNA2 and aa 534-778 of EBNA3C, regions that are not involved in binding of the viral proteins to RBP-Jkappa. The DP103-cDNA encodes a protein of 824 aa that harbors all of the common DEAD box motifs. Monoclonal antibodies raised against DP103 detect a protein of 103 kDa in mammalian cells that resides in high molecular weight complexes in vivo. We have detected an ATPase activity intrinsic to or closely associated with DP103. By subcellular fractionation, we find DP103 in both a soluble nuclear fraction as well as in the insoluble skeletal fraction. Whereas the protein and its mRNA are uniformly expressed in all tested cell lines, we observed differential expression of the mRNA in normal human tissues.
Grundhoff
A T
AT
Institut für Medizinische Mikrobiologie und Hygiene, Abteilung Virologie, Universitätskliniken des Saarlandes, Germany.
Kremmer
E
E
Türeci
O
O
Glieden
A
A
Gindorf
C
C
Atz
J
J
Mueller-Lantzsch
N
N
Schubach
W H
WH
Grässer
F A
FA
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D055541:DEAD Box Protein 20; D053487:DEAD-box RNA Helicases; D018076:DNA, Complementary / Q000302:isolation & purification; D019309:Epstein-Barr Virus Nuclear Antigens / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D020365:RNA Helicases / Q000737:chemistry* / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D019105:Random Amplified Polymorphic DNA Technique; D051381:Rats; D018411:Spodoptera; D013601:T-Lymphocytes / Q000378:metabolism
D018076:DNA, Complementary; D019309:Epstein-Barr Virus Nuclear Antigens; D012333:RNA, Messenger; D000251:Adenosine Triphosphatases; C120168:DDX20 protein, human; D055541:DEAD Box Protein 20; D053487:DEAD-box RNA Helicases; D020365:RNA Helicases