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Journal of Virology, April 1999, p. 3404-3409, Vol. 73, No. 4
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Activation of Baculovirus Very Late Promoters by Interaction with Very Late Factor 1

Song Yang1 and Lois K. Miller1,2,*

Departments of Genetics1 and Entomology,2 University of Georgia, Athens, Georgia 30602

Received 22 September 1998/Accepted 5 January 1999

Very late factor 1 (VLF-1) of Autographa californica multicapsid nuclear polyhedrosis virus (AcMNPV) activates the transcription of two genes, polyhedrin (polh) and p10, during the final, occlusion-specific phase of infection. Using transient expression assays responsive to VLF-1, we identified linker scan mutations in the polh and p10 promoters which abolished or weakened the ability of the promoters to respond to stimulation by VLF-1. These mutations were located between the transcriptional and translational initiation sites, a region previously shown to be essential for the burst of expression during the very late phase. Addition of partially purified, epitope-tagged VLF-1 to DNA encompassing this "burst sequence" resulted in a shift in the gel electrophoretic mobility of the DNA, indicating that VLF-1 forms a complex with DNA. Addition of an antibody specific for the epitope tag of VLF-1 decreased the mobility of the DNA further, confirming the presence of VLF-1 in the complex. DNase I footprint assays revealed that VLF-1 partially purified from either insect cells or bacterial cells interacted with the burst sequences of both the polh and p10 very-late promoters. Linker scan mutations within the burst sequences severely impaired interaction between VLF-1 and the promoters. We propose that VLF-1 transactivates the polh and p10 promoters by interacting with the burst sequences.

* Corresponding author. Mailing address: Department of Entomology, 413 Biological Sciences Bldg., The University of Georgia, Athens, GA 30602. Phone: (706) 542-2294. Fax: (706) 542-2279. E-mail: miller{at}arches.uga.edu.

Journal of Virology, April 1999, p. 3404-3409, Vol. 73, No. 4
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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