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J Biol Chem, Vol. 274, Issue 29, 20738-20743, July 16, 1999
From the The molecular mechanisms directing the highly
restricted expression pattern of murine ameloblastin were characterized
by cloning and functional analysis of the ameloblastin promoter. The
transcription start site, mapped by primer extension, was located 19 base pairs (bp) 5' of the published cDNA. The promoter was analyzed
in a mouse ameloblast-like cell line (LS8) and was compared with
promoter activity in primary gingival fibroblasts and pulp fibroblasts. Sequential 5'-deletion mutants encompassing DNA sequences from
Cloning and Characterization of the Murine Ameloblastin
Promoter
,
University of Michigan, School of Dentistry,
Ann Arbor, Michigan 48109-1078, the § Craniofacial
Developmental Biology and Regeneration Branch, NIDCR, National
Institutes of Health, Bethesda, Maryland 20892, and the
¶ University of Southern California School of Dentistry, Center
for Craniofacial Molecular Biology, Los Angeles, California 90033
1616
to 781 bp exhibited high promoter activity in LS8 cells, whereas the
promoter activity decreased to 50% of the full-length construct in the
781- and 477-bp regions. The 217-bp promoter region regained
promoter activity that approached the activity of the full-length
promoter construct, suggesting that both positive and negative
cis-acting regions may be involved in ameloblastin transcriptional regulation. Activity of the ameloblastin promoter in
gingival and pulp fibroblasts was minimal and ranged from 8 to 30% of
the activity in ameloblast-like cells. Several DNA-protein complexes
were formed between functionally important promoter fragments and
nuclear extracts from LS8 cells. The inactivity of promoter constructs
in pulp and gingival fibroblasts as well as the absence of similar
DNA-protein complexes from these cells suggest that regulatory regions
of the murine ameloblastin promoter may function in a cell-specific manner.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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