VZV Foundation - A Decade Of Leadership In The Fight Against VZV Infections

	Oral Presentation Abstracts: 33 [33] CELLULAR FACTORS AND IE62 ACTIVATION OF VZV PROMOTERS *W. T. Ruyechan H. He, H. Peng, M. Yang, and J. Hay Department of Microbiology, SUNY at Buffalo, Buffalo, New York The cellular transcription factors Sp1 and USF bind to a cis-acting element within the VZV gI promoter in magnetic bead capture assays. Both SP1 and USF sites must be present for activation of the gI promoter by the VZV major transactivating protein, IE62. Mutation of the binding site of either factor eliminates binding of that factor and reduces reporter gene expression to basal levels. The binding of Sp1 and USF is not dependent upon the presence of IE62 or other VZV proteins. In contrast, IE62 is required for efficient binding of the TATA-binding protein and TAF130 to the gI promoter. These data indicate that the IE62 protein recruits specific elements of TFIID to the promoter. Footprinting and mutation analysis have identified a putative binding site for the IE62 protein within the gI promoter. We have also begun an analysis of the bidirectional promoter regulating expression of VZV ORFs 28 and 29. This element also contains USF and Sp1 consensus binding sites. These experiments are aimed at developing a general model for IE62 activation of VZV promoters. Corresponding Author: W. T. Ruyechan, Ph.D., Professor, Department of Microbiology, State University of New York at Buffalo, Buffalo, NY 14214, USA

[33]

CELLULAR FACTORS AND IE62 ACTIVATION OF VZV PROMOTERS

*W. T. Ruyechan H. He, H. Peng, M. Yang, and J. Hay
Department of Microbiology, SUNY at Buffalo, Buffalo, New York

The cellular transcription factors Sp1 and USF bind to a cis-acting element within the VZV gI promoter in magnetic bead capture assays. Both SP1 and USF sites must be present for activation of the gI promoter by the VZV major transactivating protein, IE62. Mutation of the binding site of either factor eliminates binding of that factor and reduces reporter gene expression to basal levels. The binding of Sp1 and USF is not dependent upon the presence of IE62 or other VZV proteins. In contrast, IE62 is required for efficient binding of the TATA-binding protein and TAF130 to the gI promoter. These data indicate that the IE62 protein recruits specific elements of TFIID to the promoter. Footprinting and mutation analysis have identified a putative binding site for the IE62 protein within the gI promoter. We have also begun an analysis of the bidirectional promoter regulating expression of VZV ORFs 28 and 29. This element also contains USF and Sp1 consensus binding sites. These experiments are aimed at developing a general model for IE62 activation of VZV promoters.

Corresponding Author: W. T. Ruyechan, Ph.D., Professor, Department of Microbiology, State University of New York at Buffalo, Buffalo, NY 14214, USA