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(Received for publication, October 21, 1994; and in revised form, May 12, 1995) From the Mounting experimental evidence suggests that the TAT protein,
released from human immunodeficiency virus-1 (HIV-1)-infected
inflammatory cells, may genetically reprogram targeted cells within a
localized environment to develop highly vascularized tumors of
mesenchymal origin. The fibroblast growth factor (FGF) family of
polypeptides has gained general acceptance as initiators of
angiogenesis and functions as potent mitogens for mesoderm-derived
cells. To evaluate a potential biological relationship between TAT and
acidic FGF (FGF-1), primary murine embryonic fibroblasts either were
transfected with the viral transactivator or were transduced
(retrovirally mediated) with a secreted, chimeric form of the human
polypeptide growth factor, human stomach tumor/Kaposi's sarcoma
(hst/KS)FGF-1. Reverse transcriptase-polymerase chain reaction, Western
blotting, in situ immunohistochemical, heparin affinity, DNA
synthesis, and transient transfection techniques were used to confirm
expression, localization, and functionality of the transgenes. Both
transfected and transduced cells constitutively expressing either TAT
or (hst/KS)FGF-1 adopted a transformed phenotype, maintained aggressive
growth behavior, and demonstrated both induction of FGF-specific
phosphotyrosyl proteins and nuclear association of FGF-1 and FGF-1
receptor. Increased levels of endogenous, murine FGF-1 mRNA (reverse
transcriptase-polymerase chain reaction) and protein (immunoblot
analysis) were apparent in both (hst/KS)FGF-1- and TAT-transformed
cells. Medium conditioned by (hst/KS)FGF-1-transduced cells contained
steady-state levels of biologically active FGF-1 which exhibited a
representative molecular weight. Limited sodium dodecyl
sulfate-polyacrylamide gel electrophoretic analysis of the conditioned
medium from TAT-transformed cells demonstrated the appearance of FGF-1
as latent, high molecular weight complexes requiring reducing agents to
activate full biological activity. Collectively, these results suggest
that TAT induces the expression and secretion of FGF-1, which may be
potentially relevant to the pathophysiological development of
AIDS-Kaposi's sarcoma.
Volume 270,
Number 29,
Issue of July 21, pp. 17457-17467, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
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