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Previous observations have shown that binding of growth hormone
to its receptor leads to activation of transcription factors via a
mechanism involving phosphorylation on tyrosine residues. In order to
establish whether the prolactin-activated transcription factor Stat 5
(mammary gland factor) is also activated by growth hormone, nuclear
extracts were prepared from COS-7 cells transiently expressing
transfected Stat 5 and growth hormone receptor cDNA. Gel
electrophoresis mobility shift analyses revealed the growth
hormone-dependent presence of specific DNA-binding proteins in these
extracts. The complexes formed could be supershifted by polyclonal
anti-Stat 5 antiserum. In other experiments nuclear extracts from
growth hormone-treated Chinese hamster ovary cells stably expressing
transfected growth hormone receptor cDNA and liver from growth
hormone-treated hypophysectomized rats were used for gel
electrophoresis mobility shift analyses. These also revealed the
presence of specific DNA-binding proteins sharing antigenic
determinants with Stat 5. Stat 5 cDNA was shown to be capable of
complementing the growth hormone-dependent activation of transcription
of a reporter gene in the otherwise unresponsive COS-7 cell line. This
complementation was dependent on the presence of Stat 5 tyrosine 694,
suggesting a role for phosphorylation of this residue in growth
hormone-dependent activation of DNA-binding and transcription.
Volume 270,
Number 16,
Issue of April 21, pp. 9448-9453, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
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