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J Biol Chem, Vol. 274, Issue 46, 32619-32630, November 12, 1999
Binding Protein-1 (LTBP-1) in a Cell Type-specific Manner
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From the Latent transforming growth factor- Department of Virology, The Haartman
Institute, University of Helsinki, FIN-00014 Helsinki, Finland
(TGF-)-binding proteins (LTBPs) are components of the extracellular
matrix and large latent TGF- complexes are secreted by various
cells. Human LTBP-1 is known to exist in different forms. LTBP-1L
(long) has an amino-terminal extension, which is not found in the
smaller LTBP-1S isoform. To study the formation and transcriptional
regulation of LTBP-1S and LTBP-1L isoforms, we
determined the nucleotide sequences of their 5'-flanking regions. The
upstream regions of both isoforms are devoid of TATA boxes but contain
other putative binding sites for several transcription factors. Genomic
sequencing revealed that LTBP-1L transcript is
alternatively spliced to an internal splice acceptor inside exon 1 of
LTBP-1S and thus defined the genomic organization of the
isoforms. Reporter gene analysis of upstream regions indicated the
presence of independent, functional promoters, which regulate the
transcription of the isoforms by cell-specific manner. Deletion
analyses of the promoter regions revealed specific elements modulating
their basal and cell type-specific expression. In SV-40
virus-transformed WI-38 lung fibroblasts a regulatory element repressed
the transcription of LTBP-1S by a cell-specific manner. In
amniotic epithelial cells, transcription of the LTBP-1S
reporter gene construct was down-regulated by a distal upstream
element. mRNA levels of the isoforms of LTBP-1 were
stimulated in response to TGF-1 in WI-38 cells. However, since
TGF-1 failed to stimulate the transcription of LTBP-1
reporter gene constructs, TGF-1 may mediate the induction of the
isoforms by post-transcriptional mechanisms. Chromosomal
localization of the LTBP-1 gene was refined to
2p22-24.
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