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J Biol Chem, Vol. 273, Issue 42, 27524-27530, October 16, 1998

A Vaccinia Virus Late Transcription Factor with Biochemical and Molecular Identity to a Human Cellular Protein

Sajeevani K. GunasingheDagger , Alan E. Hubbs§, and Cynthia F. WrightDagger

From the Dagger  Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, South Carolina 29425 and the § Department of Cellular Pathology, Armed Forces Institute of Pathology, Washington, D. C. 20306-6000

A factor designated VLTF-X is required to support vaccinia virus late transcription in vitro. It has been found that a late promoter DNA binding activity cochromatographs and cosediments with VLTF-X activity. Current experiments show that VLTF-X activity is present in a variety of uninfected mammalian cell types and is indistinguishable from that recovered from infected cells based upon several criteria. VLTF-X activity from both sources displays the same purification profile over phosphocellulose and DNA affinity resins and has the same sedimentation coefficient. In addition, the factors purified from both infected and uninfected cells form protein-DNA complexes of identical electrophoretic mobility in the presence of vaccinia virus late promoter-containing DNA. The affinity of these factors for the late promoter probes is identical and late promoter-specific based on competition experiments. Moreover, VLTF-X purified from both sources bound to late promoter-containing DNA in the presence or absence of MgCl2 and ATP and formed complexes resistant to heat inactivation. These experiments offer proof that vaccinia virus factor VLTF-X is a host cell protein that supports transcription of the viral late genes.

Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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