Expression of the osteonectin gene potentially controlled by multiple cis- and trans-acting factors in cultured bone cells.
The cis-acting regulatory elements of the osteonectin gene have been studied using a chloramphenicol acetyltransferase (CAT) promoter assay in osteonectin-expressing and nonexpressing cultured cells. When various stretches of the promoter were transiently transfected into fetal bovine bone cells, a positive element was detected in the DNA located between bases -504 and 11 (1 being the start of transcription) and a negative element between bases -900 and -504. The positive element of the promoter also conferred preferential expression of the gene, showing more activity in cells with higher levels of osteonectin mRNA expression. A 1.2 kb fragment of intron 1 displayed a negative effect on CAT expression when inserted 5' to the promoter. An additional regulatory element was found in DNA encoding exon 1, which significantly influenced expression of the gene in fetal bovine bone cells. Gel shift analysis using positive genomic elements located 5' to the start of transcription indicated that one of the nuclear proteins that interacts with the osteonectin promoter may be related to the transcription factor AP2.
Dominguez
P
P
Bone Research Branch, National Institute of Dental Research, National Institutes of Health, Bethesda, Maryland.
Ibaraki
K
K
Robey
P G
PG
Hefferan
T E
TE
Termine
J D
JD
Young
M F
MF
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D001842:Bone and Bones / Q000166:cytology* / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase; D004268:DNA-Binding Proteins / Q000235:genetics; D005091:Exons; D005786:Gene Expression Regulation; D008969:Molecular Sequence Data; D015676:Osteonectin / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D050656:Transcription Factor AP-2; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D015676:Osteonectin; D050656:Transcription Factor AP-2; D014157:Transcription Factors; D015500:Chloramphenicol O-Acetyltransferase