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J. Biol. Chem., Vol. 276, Issue 21, 18282-18289, May 25, 2001
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From the § CIHR Group in Periodontal Physiology and the
Osteogenic differentiation involves a cascade of
coordinated gene expression that regulates cell proliferation and
matrix protein formation in a defined temporo-spatial manner. Here we have used differential display to identify a novel zinc finger transcription factor (AJ18) that is induced during differentiation of
bone cells in vitro and in vivo. The 64-kDa
protein, encoded by a 7- kilobase mRNA, contains a
Krüppel-associated box (KRAB) domain followed by 11 successive
C2H2 zinc finger motifs. AJ18 mRNA, which
is also expressed in kidney and brain, is developmentally regulated in
embryonic tibiae and calvariae, with little expression in neonate and
adult animals. During osteogenic differentiation in vitro
AJ18 mRNA is expressed as cells approach confluence and declines as
bone formation occurs. Using bacterially expressed, His-tagged AJ18 in
a target detection assay, we identified a consensus binding sequence of
5'-CCACA-3', which forms part of the consensus element for
Runx2, a master gene for osteogenic differentiation. Overexpression of AJ18 suppressed Runx2-mediated transactivation of an
osteocalcin promoter construct in transient transfection assays and reduced alkaline phosphatase activity in bone morphogenetic protein-induced C3H10T1/2 cells. These studies, therefore, have identified a novel zinc finger transcription factor in bone that can
modulate Runx2 activity and osteogenic differentiation.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF321874.
Characterization of a Novel KRAB/C2H2
Zinc Finger Transcription Factor Involved in Bone Development*
¶,
Department of Biochemistry, University of Toronto,
Toronto, Ontario M5S 3E2, Canada
*
These studies were supported by a grant from the Canadian
Institute of Health Research (MOP 37786).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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