AcMNPV AC16 (DA26, BV/ODV-E26) regulates the levels of IE0 and IE1 and binds to both proteins via a domain located within the acidic transcriptional activation domain.
IE0 and IE1 are the primary viral regulatory proteins of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) involved in the transactivation of early genes, stimulation of late gene expression, and viral DNA replication. The protein interactions required for IE0 or IE1 to achieve these varied roles are not well defined, so to identify proteins that interact with IE0 and IE1, tandem affinity purification (TAP) and LC-MS/MS was used. Analysis of purified proteins identified AC16 (DA26, BV/ODV-E26) from TAP tagged IE0 virus infected Sf9 cells. Co-immunoprecipitation confirmed that AC16 interacts with both IE0 and IE1 and yeast 2-hybrid analysis mapped the domain required for interaction with AC16. Mutation of the AC16 binding domain enhanced BV production by viruses expressing only IE0 but had no effect if only IE1 is expressed. An ac16 deletion virus was constructed and was shown not to affect the temporal expression of IE0 and IE1; however the relative level of IE0 to IE1 was significantly increased.
Nie
Yingchao
Y
Plant Science, Faculty of Land and Food Systems, University of British Columbia, Vancouver, B.C., Canada V6T 1Z4.
Fang
Minggang
M
Theilmann
David A
DA
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D017353:Gene Deletion; D015967:Gene Expression Regulation, Viral / Q000235:genetics*; D005822:Genetic Vectors; D017874:Immediate-Early Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D047468:Immunoprecipitation; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D018411:Spodoptera / Q000821:virology; D015534:Trans-Activators / Q000737:chemistry / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014760:Viral Fusion Proteins / Q000235:genetics; D014779:Virus Replication / Q000235:genetics
C069183:Ac-IE-1 protein, Autographa californica nucleopolyhedrovirus; C110347:AcOrf-16 protein, Autographa californica nucleopolyhedrovirus; D017874:Immediate-Early Proteins; D015534:Trans-Activators; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins; C450080:ie0 protein, baculovirus