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

	Oral Presentation Abstracts: 62 [62] ANALYSIS OF THE VZV ORF63 PROTEIN *J. Lynch (1), M. Spengler (1), T. Kenyon (2), C. Grose (2), J. Hay (1) and W. T. Ruyechan (1) (1)Department of Microbiology, SUNY at Buffalo, Buffalo, New York 14214; (2)Departments of Microbiology and Pediatrics, University of Iowa, Iowa City, IA The VZV ORF63/70 genes encode a protein with a predicted mass of 30.5 kDa although the species identified in infected cells exhibits an apparent molecular weight of 45 kDa.. The ORF63 protein is expressed early in lytic infection and is also incorporated into the virus particle as a tegument protein. Recent work in Dr. Ann Arvin's laboratory indicates that the ORF63 protein is required for growth in tissue culture. Several previous studies have shown that ORF63 mRNA and protein are abundantly expressed during latency. Currently, however, little is known concerning ORF63 function. ORF63 is predicted to be a phospho protein with consensus sites for casein kinase II and protein kinase C [PKC]. Using recombinant fusion proteins we have shown that the ORF63 protein is phosphorylated by casein kinase II. We have also shown that the ORF63 protein is heavily phosphorylated by the VZV ORF47 kinase. Elisa-binding assays and immunoblotting have shown that the ORF63 protein can interact directly with the VZV IE62 major transactivator.CAT assays have been performed to characterize a functional interaction between IE62 and IE63. Our current data indicate that the ORF63 protein has no intrinsic transactivating activity but can boost the transactivation of the VZV gI promoter by IE62. Corresponding Author: W. T. Ruyechan, Ph.D., Professor, Department of Microbiology, State University of New York at Buffalo, Buffalo, NY 14214, USA

[62]

ANALYSIS OF THE VZV ORF63 PROTEIN

*J. Lynch (1), M. Spengler (1), T. Kenyon (2), C. Grose (2), J. Hay (1) and
W. T. Ruyechan (1)
(1)Department of Microbiology, SUNY at Buffalo, Buffalo, New York 14214;
(2)Departments of Microbiology and Pediatrics, University of Iowa, Iowa City, IA

The VZV ORF63/70 genes encode a protein with a predicted mass of 30.5 kDa although the species identified in infected cells exhibits an apparent molecular weight of 45 kDa.. The ORF63 protein is expressed early in lytic infection and is also incorporated into the virus particle as a tegument protein. Recent work in Dr. Ann Arvin's laboratory indicates that the ORF63 protein is required for growth in tissue culture. Several previous studies have shown that ORF63 mRNA and protein are abundantly expressed during latency. Currently, however, little is known concerning ORF63 function. ORF63 is predicted to be a phospho protein with consensus sites for casein kinase II and protein kinase C [PKC]. Using recombinant fusion proteins we have shown that the ORF63 protein is phosphorylated by casein kinase II. We have also shown that the ORF63 protein is heavily phosphorylated by the VZV ORF47 kinase. Elisa-binding assays and immunoblotting have shown that the ORF63 protein can interact directly with the VZV IE62 major transactivator.CAT assays have been performed to characterize a functional interaction between IE62 and IE63. Our current data indicate that the ORF63 protein has no intrinsic transactivating activity but can boost the transactivation of the VZV gI promoter by IE62.

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