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J Biol Chem, Vol. 273, Issue 22, 13768-13775, May 29, 1998
From the The human T-cell leukemia virus type I Tax
protein forms a ternary complex on DNA in association with a host
factor, the cyclic AMP response element-binding protein (CREB). An
understanding of the precise geometry of this complex has been elusive.
We have used photocross-linking to investigate Tax-DNA contacts. Our
data show that Tax contacts the DNA at two symmetric positions 14 nucleotides apart on either side of the Tax responsive element. The
presence of symmetric, widely separated regions of contact suggests
that at least two molecules of Tax are present in the complex. Mapping the contacts onto a three-dimensional model of the CREB-DNA binary complex shows that they lie on the same face of the DNA near the regions where the N termini of the CREB bZIP domains enter the major
groove. This location correlates well with previous evidence that CREB
amino acid residues immediately N-terminal to the bZIP domain are
crucial for the formation of the ternary complex. The limited number of
cross-links observed suggests that contacts are primarily with the
phosphate backbone and does not support the idea that a major
structural element of the Tax protein inserts into the major or minor
grooves of the DNA.
Specific Regions of Contact between Human T-cell Leukemia Virus
Type I Tax Protein and DNA Identified by Photocross-linking
§ and
Gene Regulation Program, Institute of
Molecular Medicine and Genetics, Medical College of Georgia,
Augusta, Georgia 30912 and the § Department of Chemistry and
Biochemistry, University of Colorado,
Boulder, Colorado 80309-0215
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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