Use of an N-terminal half truncated IE1 as an antagonist of IE1, an essential regulatory protein in baculovirus.
An immediate-early gene product of baculovirus, IE1, is essential for viral gene expression and for viral DNA replication. It has been demonstrated for Autographa californica nuclear polyhedrosis virus (AcNPV) that the C-terminal region of IE1 is required for dimerization. And the acidic N-terminal region of IE1 has been identified as the activation domain. We constructed an N-terminal 267 amino acid (a.a.) truncated mutant of Bombyx mori nuclear polyhedrosis virus (BmNPV) IE1, which was defective as a transactivator of a viral early gene (p35) promoter. We then examined possible IE1 antagonistic functions of this defective IE1, IE1TN, in BmNPV-infected cells. A transient expression experiment demonstrated that IE1TN strongly repressed the activation of the hr5-dependent p35 promoter derived from BmNPV infection. In addition, DpnI assay elucidated an inhibitory effect of IE1TN on the hr5-dependent replication of plasmid in BmN cells induced by NPV infection. A marked reduction in the production of virus was observed when the BmN cells were infected with BmNPV after transfection with IE1TN-expression plasmids. These results suggested that IE1TN could act as an IE1 antagonist in silkworm cells infected with BmNPV. We then analyzed the ability of IE1TN to inhibit the multiplication of BmNPV using transgenic silkworms. The BmNPV-resistance of the transgenic silkworms was very weak, suggesting insufficient expression of the transgene product, IE1TN.
Yamada
Yoji
Y
Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University, Kita 9, Nishi 9, Kita-ku, Sapporo 060-8589, Japan.
Matsuyama
Takahiro
T
Quan
Guo-Xing
GX
Kanda
Toshio
T
Tamura
Toshiki
T
Sahara
Ken
K
Asano
Shin-ichiro
S
Bando
Hisanori
H
D000818:Animals; D030801:Animals, Genetically Modified; D012831:Bombyx / Q000235:genetics / Q000821:virology*; D002478:Cells, Cultured; D004261:DNA Replication; D004268:DNA-Binding Proteins; D017353:Gene Deletion; D015967:Gene Expression Regulation, Viral; D017874:Immediate-Early Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics*; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000472:pathogenicity*; D011401:Promoter Regions, Genetic; D015534:Trans-Activators / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics*; D015533:Transcriptional Activation; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
C069183:Ac-IE-1 protein, Autographa californica nucleopolyhedrovirus; D004268:DNA-Binding Proteins; D017874:Immediate-Early Proteins; D015534:Trans-Activators; D014764:Viral Proteins; C081618:p35 protein, Nucleopolyhedrovirus