Major product pp43 of human cytomegalovirus U(L)112-113 gene is a transcriptional coactivator with two functionally distinct domains.
Human cytomegalovirus U(L)112-113 encodes four phosphoproteins, pp84, pp50, pp43, and pp34, with common amino-termini. A previous report by Kerry et al. (J. Virol. 70, 373-382, 1996) demonstrated that U(L)112-113 products activate U(L)54 promoter in cooperation with immediate-early (IE) proteins. In this study, we identified a domain required for transcriptional activation in the pp43 protein, which consisted of two distinct regions: domain I (amino acids 272-296) and domain II (amino acids 297-306). Domain I contained two long glycine stretches, and domain II was a short proline-containing region. Both of domains were required for IE2-dependent transcriptional activation. The pp43 mutant that had domain I but lacked domain II acted as a dominant negative mutant and suppressed most of the IE2-dependent activation, indicating the importance of coactivation by pp43 in this transcriptional activation. The major protein pp43 also weakly activated the promoter through IR1 element in a manner independent of IE2. Only domain I was required for this IE2-independent activation. These domains were common in pp84, pp50, and pp43 but did not exist in pp34, which did not activate transcription alone. These results suggest that the major product, pp43, of U(L)112-113 has two functionally distinct domains and plays an important role in mediating IE2-dependent transcriptional activation.
Division of Virology and Immunology, Medical Research Institute, Tokyo Medical and Dental University, Yushima 1-5-45, Tokyo, Bunkyo-ku, 113-8510, Japan.
J
J
Li
T
T
Yamamoto
K
K
Ohtsubo
M
M
Shirakata
K
K
Hirai
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002460:Cell Line; D003587:Cytomegalovirus / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D019913:Nuclear Localization Signals / Q000235:genetics; D010750:Phosphoproteins / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011487:Protein Conformation; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D014759:Viral Envelope Proteins; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism
D004268:DNA-Binding Proteins; C069186:IE2 protein, Cytomegalovirus; D017874:Immediate-Early Proteins; D008562:Membrane Glycoproteins; D019913:Nuclear Localization Signals; D010750:Phosphoproteins; D015534:Trans-Activators; C077406:UL115 protein, Human herpesvirus 5; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C090854:cytomegalovirus phosphoprotein 43kDa; C071440:glycoprotein H, Cytomegalovirus; C499358:glycoprotein H, Human cytomegalovirus; C439068:glycoprotein O, cytomegalovirus