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Vol. 11, Issue 6, 2007-2018, June 2000
2 Transcriptional Activation Domain of
Glucocorticoid Receptor with a Novel Steroid Receptor Coactivator,
Hic-5, Which Localizes to Both Focal Adhesions and the Nuclear Matrix
and
*Department of Pathology and Department of Biochemistry and
Molecular Biology, University of Southern California, Los Angeles,
California 90089; and Hic-5 (hydrogen peroxide-inducible clone-5) is a focal adhesion
protein that is involved in cellular senescence. In the present study,
a yeast two-hybrid screen identified Hic-5 as a protein that interacts
with a region of the glucocorticoid receptor that includes a nuclear
matrix-targeting signal and the Department of Biological Sciences
and Department of Neuroscience and Pharmacology,
University of Pittsburgh, Pittsburgh, Pennsylvania 15260
2 transcriptional activation
domain. In transiently transfected mammalian cells, overexpression of
Hic-5 potentiated the activation of reporter genes by all steroid
receptors, excluding the estrogen receptor. The activity of the
estrogen receptor and the thyroid hormone receptor was stimulated by
Hic-5 in the presence but not in the absence of coexpressed coactivator
GRIP1. In biochemical fractionations and indirect immunofluorescence
assays, a fraction of endogenous Hic-5 in REF-52 cells and transiently
expressed Hic-5 in Cos-1 cells was associated with the nuclear matrix.
The C-terminal region of Hic-5, which contains seven zinc fingers
arranged in four LIM domains, was required for interaction with focal
adhesions, the nuclear matrix, steroid receptors, and the 2 domain
of glucocorticoid receptor. The N-terminal region of Hic-5 possesses a
transcriptional activation domain and was essential for the coactivator
activity of Hic-5. Given the coexisting cytoplasmic and nuclear
distributions of Hic-5 and its role in steroid receptor-mediated
transcriptional activation, it is proposed that Hic-5 might transmit
signals that emanate at cell attachment sites and regulate
transcription factors, such as steroid receptors.
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