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(Received for publication, November 5, 1996, and in revised form, December 20, 1996)
From the The LIM domain protein rhombotin-2
(RBTN-2/TTG-2/LMO2) is involved in many processes, including
leukemogenesis and erythropoiesis. It is thought that the principle
role of RBTN-2 in these processes is to regulate transcription. To
examine the potential for RBTN-2 to modulate transcription, we
constructed RBTN-2/GAL4 DNA-binding domain fusion proteins and measured
their ability to activate transcription of a reporter gene construct.
From these studies we identified a transcription activation domain
within the NH2 terminus of RBTN-2. This activation domain
was further localized within a proline-rich 19-amino acid region. A
second activation domain of 11 amino acids was also identified. This
domain was located within the COOH terminus of RBTN-2, and functioned
in mammalian cells but not in yeast. Furthermore, the two LIM domains of RBTN-2 were shown to function as transcription repression domains. Each individual LIM domain acted as an independent transcription repression domain on a heterologous activation domain. However, in
context of full-length RBTN-2, the LIM domains selectively repressed
the NH2-terminal activation domain, but had no effect on
the COOH-terminal domain. Overall, these results demonstrate that the
T-cell oncogene RBTN-2 is a complex transcription factor possessing
multiple transcription regulatory modules, including two activation
domains and two repression domains.
Volume 272, Number 9,
Issue of February 28, 1997
pp. 5594-5599
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
§
,
and§
Department of Virology and Molecular
Biology, St. Jude Children's Research Hospital, Memphis, Tennessee
38101 and the § Department of Pathology, University of
Tennessee, Memphis, Tennessee 38163
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