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J Biol Chem, Vol. 273, Issue 16, 9457-9464, April 17, 1998
,
,
From the University of Texas Health Science Center at San
Antonio, Dental School, San Antonio, Texas
78284-7888, the Our laboratory has reported that two
major noncollagenous dentin proteins, dentin sialoprotein and dentin
phosphoprotein, are specific cleavage products of a larger precursor
protein termed dentin sialophosphoprotein (MacDougall, M.,
Simmons, D., Luan, X., Nydegger, J., Feng, J. Q., and Gu,
T. T. (1997) J. Biol. Chem. 272:835-842). To
confirm our single gene hypothesis and initiate in vitro
promoter studies, we have characterized the structural organization of
the mouse dentin sialophosphoprotein gene. This gene has a
transcription unit of ~9.4 kilobase pairs and is organized into 5 exons and 4 introns. Exon 1 contains a noncoding 5' sequence, and exon
2 contains the transcriptional start site, signal peptide, and first
two amino acids of the NH2 terminus. Exons 3 and 4 contain coding information for 29 and 314 amino acids, respectively. The remainder of the coding information and the untranslated 3' region are
contained in exon 5. Chromosomal mapping localized the gene to mouse
chromosome 5q21 in close proximity to other dentin/bone matrix genes.
Computer analysis of the promoter proximal 1.6-kilobase pair sequence
revealed a number of potentially important cis-regulatory sequences; these include the recognition elements of AP-1, AP-2, Msx-1,
serum response elements, SP-1, and TCF-1. In vitro studies showed that the DSPP promoter is active in an odontoblast cell line,
MO6-G3, with basal activity mapped to
Gene Targeting Research and
Core Facility, NIDR, National Institutes of Health, Bethesda, Maryland
20892, the § University of Texas Health Science Center
Houston Dental Branch, Houston, Texas 78284, and
¶ NIAID, National Institutes of Health,
Bethesda, Maryland 20892
95 bp. Two potential enhancer
and suppresser elements were identified in the regions between
1447
and
791 bp and
791 and
95 bp, respectively. The structural
organization of the dentin sialophosphoprotein gene confirms our
finding that both dentin sialoprotein and dentin phosphoprotein are
encoded by a single gene with a continuous open reading frame.
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