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(Received for publication, February 19, 1997, and in revised form, August 11, 1997)
From the Transcriptional up-regulation of the
c-sis/platelet-derived growth factor-B (PDGF-B)
proto-oncogene by the Tax protein of human T-cell leukemia virus type 1 has been implicated as one possible mechanism of cellular
transformation by human T-cell leukemia virus type 1. In previous work,
we identified an essential site in the c-sis/PDGF-B
promoter, Tax-responsive element 1 (TRE1), necessary for
transactivation by Tax. We also identified Sp1, Sp3, and NGFI-A/Egr-1
as the primary nuclear transcription factors binding to TRE1 which
mediate Tax responsiveness. In the present work, we have investigated
the mechanism(s) whereby Tax transactivates the
c-sis/PDGF-B proto-oncogene. In vitro
transcription assays showed that Tax was able to significantly
increase the transcriptional activity of a template containing the
Volume 272, Number 43,
Issue of October 24, 1997
pp. 27411-27421
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
The Tax Protein of Human T-cell Leukemia Virus Type 1 Mediates
the Transactivation of the c-sis/Platelet-derived Growth
Factor-B Promoter through Interactions with the Zinc Finger
Transcription Factors Sp1 and NGFI-A/Egr-1
,
Division of Molecular Oncology, Washington
University School of Medicine, St. Louis, Missouri 63110 and the
§ McArdle Laboratory for Cancer Research, University of
Wisconsin Medical School, Madison, Wisconsin 53706
257 to +74 region of the c-sis/PDGF-B promoter.
Electrophoretic mobility shift assay analysis showed that Tax increased
the DNA binding activity of both Sp1 and NGFI-A/Egr-1 using a TRE1
probe. Analysis of Tax mutants showed that two mutants, IEXC29S and
IEXL320G, were unable to significantly transactivate the
c-sis/PDGF-B promoter. Finally, co-immunoprecipitation analysis revealed that Tax is able to stably bind to both Sp1 and
NGFI-A/Egr-1. Interestingly, co-immunoprecipitation analysis also
revealed that Tax mutant IEXC29S is unable to interact with NGFI-A/Egr-1, whereas Tax mutant IEXL320G is able to interact with
NGFI-A/Egr-1.
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