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J Biol Chem, Vol. 274, Issue 50, 35662-35667, December 10, 1999
From the Department of Biochemistry, Purdue University,
West Lafayette, Indiana 47907-1153
Vaccinia virus has a DNA genome, yet replicates
in the cytoplasmic compartment of the cell. We previously described the
identification of a cellular protein having high affinity for vaccinia
virus late promoter DNA. Sequence substitutions in the vaccinia I1L promoter were used to define a 5-nucleotide block at the transcription initiation site as essential for interaction with the protein. Within
this sequence is the recognition motif for the nuclear transcription
factor YY1. This factor regulates a multitude of cellular promoters, as
an activator of transcription, as a repressor, or as an initiator
element-binding protein. Antibodies directed against YY1 were used to
show that YY1 copurified with the vaccinia late promoter-binding
protein and was present in late promoter-protein complexes in gel
supershift assays. Bacterially expressed YY1 also bound specifically to
late promoter DNA. A dinucleotide replacement within the YY1
recognition motif directly adjacent to the transcription start site
severely reduced the affinity of YY1 for the I1L promoter in
vitro and impaired I1L promoter-dependent
transcription in vivo. The intracellular localization of
YY1 was shown by immunofluorescence microscopy to shift from primarily
nuclear to the cytoplasm after vaccinia infection. These results
indicate that YY1 has a positive role in the regulation of vaccinia
virus late gene transcription and suggest that poxviruses have adapted
cellular initiator elements as a means of regulating viral gene
expression. This is the first identifiable cellular protein implicated
in poxvirus transcription.
Transcription Factor YY1 Is a Vaccinia Virus Late Promoter
Activator*
,
*
This work was supported by a grant from the NIAID, National
Institutes of Health. This is paper number 16028 from the Purdue Agricultural Experiment Station.The costs of publication of this article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.: 765-494-0745;
Fax: 765-494-7897; E-mail: broyles@biochem.purdue.edu.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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