RING and coiled-coil domains of baculovirus IE2 are critical in strong activation of the cytomegalovirus major immediate-early promoter in mammalian cells.
In recent years, baculovirus has emerged as a tool for high-efficiency gene transfer into mammalian cells. However, the level of gene expression is often limited by the strength of the mammalian promoter used. Here, we show that the baculovirus RING protein IE2 is a strong, promiscuous trans-activator in mammalian cells, dramatically upregulating the cytomegalovirus (CMV) promoter in both Vero E6 and U-2OS cells. Further study of the cellular mechanism for the activation led to the discovery of a novel IE2 nuclear body structure which contains a high concentration of G-actin and closely associates with RNA polymerase II, PML, and SUMO1. IE2 mutagenesis studies indicated that the RING and coiled-coil domains of IE2 were necessary for nuclear body formation, as well as for strong activation of the CMV promoter in mammalian cells. Overall, this study shows that the IE2 trans-activator could significantly advance the use of baculovirus in mammalian gene transfer and protein production.
Liu
Catherine Yen-Yen
CY
Institute of Molecular Biology, Academia Sinica, Taipei 115, Taiwan, Republic of China.
Wang
Chia-Hung
CH
Hsiao
Wen-Kai
WK
Lo
Huei-Ru
HR
Wu
Carol P
CP
Chao
Yu Chan
YC
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002460:Cell Line; D002522:Chlorocebus aethiops; D003587:Cytomegalovirus / Q000235:genetics*; D015870:Gene Expression; D017781:Genes, Immediate-Early; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism*; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection; D014773:Virology / Q000379:methods*
D017874:Immediate-Early Proteins; D015534:Trans-Activators; C072773:IE2 protein, polyhedrosis virus