Coordinate occupancy of AP-1 sites in the vitamin D-responsive and CCAAT box elements by Fos-Jun in the osteocalcin gene: Model for phenotype suppression of transcription

Thomas A. Owen, Rita Bortell, Sue A. Yocum, Steven L. Smock, Meng Zhang, Cory Abate, Victoria Shalhoub, Neil Aronin, Kenneth L. Wright, Andre J van Wijnen, Janet L. Stein, Tom Curran, Jane B. Lian, Gary S. Stein

Research output: Contribution to journalArticle

173 Citations (Scopus)

Abstract

Osteocalcin, a bone-specific protein and marker of the mature osteoblast, is expressed only in nonproliferating osteoblasts in a mineralizing extracellular matrix, while type I collagen is expressed in proliferating cells. The nuclear proteins encoded by the c-fos and c-jun protooncogenes are expressed during the proliferation period of osteoblast phenotype development. We present evidence that AP-1 (HeLa cell-activating protein 1) sites residing within two promoter elements of the osteocalcin gene bind the Fos-Jun protein complex: the osteocalcin box (OC box; nucleotides -99 to -76), which contains a CCAAT motif as a central element and influences tissue-specific basal levels of osteocalcin gene transcription, and the vitamin D-responsive element (VDRE; nucleotides -462 to -440), which mediates enhancement of osteocalcin gene transcription. Gel electrophoretic mobility-shift analysis demonstrated high AP-1 binding activity in proliferating osteoblasts and dramatic changes in this activity after the down-regulation of proliferation and the initiation of extracellular-matrix mineralization in primary cultures of normal diploid osteoblasts. Methylation interference analysis established at single nucleotide resolution that purified recombinant Fos and Jun proteins bind in a sequence-specific manner to the AP-1 sites within the VDRE and OC box. Similarly, an AP-1 motif within a putative VDRE of the alkaline phosphatase gene, which is also expressed after the completion of proliferation, binds the Fos-Jun complex. These results support a model in which coordinate occupancy of the AP-1 sites in the VDRE and OC box in proliferating osteoblasts may suppress both basal level and vitamin D-enhanced osteocalcin gene transcription as well as transcription of other genes associated with osteoblast differentiation - a phenomenon we describe as phenotype suppression. This model is further supported by binding of the Fos-Jun complex at an AP-1 site in the type αI collagen promoter that is contiguous with, but not overlapping, the VDRE. Such a sequence organization in the collagen VDRE motif is compatible with vitamin D modulation of collagen but not with osteocalcin and alkaline phosphatase expression in proliferating osteoblasts.

Original languageEnglish (US)
Pages (from-to)9990-9994
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number24
StatePublished - 1990
Externally publishedYes

Fingerprint

Osteocalcin
Osteoblasts
HeLa Cells
Vitamin D
Phenotype
Genes
Proteins
Nucleotides
Collagen Type I
Extracellular Matrix
Alkaline Phosphatase
Collagen
fos Genes
Amino Acid Motifs
Nuclear Proteins
Diploidy
Protein Binding
Methylation
Down-Regulation
Gels

Keywords

  • Collagen
  • Differentiation
  • Growth control
  • Oncogenes
  • Proliferation
  • Transcription

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Coordinate occupancy of AP-1 sites in the vitamin D-responsive and CCAAT box elements by Fos-Jun in the osteocalcin gene : Model for phenotype suppression of transcription. / Owen, Thomas A.; Bortell, Rita; Yocum, Sue A.; Smock, Steven L.; Zhang, Meng; Abate, Cory; Shalhoub, Victoria; Aronin, Neil; Wright, Kenneth L.; van Wijnen, Andre J; Stein, Janet L.; Curran, Tom; Lian, Jane B.; Stein, Gary S.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 87, No. 24, 1990, p. 9990-9994.

Research output: Contribution to journalArticle

Owen, TA, Bortell, R, Yocum, SA, Smock, SL, Zhang, M, Abate, C, Shalhoub, V, Aronin, N, Wright, KL, van Wijnen, AJ, Stein, JL, Curran, T, Lian, JB & Stein, GS 1990, 'Coordinate occupancy of AP-1 sites in the vitamin D-responsive and CCAAT box elements by Fos-Jun in the osteocalcin gene: Model for phenotype suppression of transcription', Proceedings of the National Academy of Sciences of the United States of America, vol. 87, no. 24, pp. 9990-9994.
Owen, Thomas A. ; Bortell, Rita ; Yocum, Sue A. ; Smock, Steven L. ; Zhang, Meng ; Abate, Cory ; Shalhoub, Victoria ; Aronin, Neil ; Wright, Kenneth L. ; van Wijnen, Andre J ; Stein, Janet L. ; Curran, Tom ; Lian, Jane B. ; Stein, Gary S. / Coordinate occupancy of AP-1 sites in the vitamin D-responsive and CCAAT box elements by Fos-Jun in the osteocalcin gene : Model for phenotype suppression of transcription. In: Proceedings of the National Academy of Sciences of the United States of America. 1990 ; Vol. 87, No. 24. pp. 9990-9994.
@article{4dfb5ed15c204facb45890a593757001,
title = "Coordinate occupancy of AP-1 sites in the vitamin D-responsive and CCAAT box elements by Fos-Jun in the osteocalcin gene: Model for phenotype suppression of transcription",
abstract = "Osteocalcin, a bone-specific protein and marker of the mature osteoblast, is expressed only in nonproliferating osteoblasts in a mineralizing extracellular matrix, while type I collagen is expressed in proliferating cells. The nuclear proteins encoded by the c-fos and c-jun protooncogenes are expressed during the proliferation period of osteoblast phenotype development. We present evidence that AP-1 (HeLa cell-activating protein 1) sites residing within two promoter elements of the osteocalcin gene bind the Fos-Jun protein complex: the osteocalcin box (OC box; nucleotides -99 to -76), which contains a CCAAT motif as a central element and influences tissue-specific basal levels of osteocalcin gene transcription, and the vitamin D-responsive element (VDRE; nucleotides -462 to -440), which mediates enhancement of osteocalcin gene transcription. Gel electrophoretic mobility-shift analysis demonstrated high AP-1 binding activity in proliferating osteoblasts and dramatic changes in this activity after the down-regulation of proliferation and the initiation of extracellular-matrix mineralization in primary cultures of normal diploid osteoblasts. Methylation interference analysis established at single nucleotide resolution that purified recombinant Fos and Jun proteins bind in a sequence-specific manner to the AP-1 sites within the VDRE and OC box. Similarly, an AP-1 motif within a putative VDRE of the alkaline phosphatase gene, which is also expressed after the completion of proliferation, binds the Fos-Jun complex. These results support a model in which coordinate occupancy of the AP-1 sites in the VDRE and OC box in proliferating osteoblasts may suppress both basal level and vitamin D-enhanced osteocalcin gene transcription as well as transcription of other genes associated with osteoblast differentiation - a phenomenon we describe as phenotype suppression. This model is further supported by binding of the Fos-Jun complex at an AP-1 site in the type αI collagen promoter that is contiguous with, but not overlapping, the VDRE. Such a sequence organization in the collagen VDRE motif is compatible with vitamin D modulation of collagen but not with osteocalcin and alkaline phosphatase expression in proliferating osteoblasts.",
keywords = "Collagen, Differentiation, Growth control, Oncogenes, Proliferation, Transcription",
author = "Owen, {Thomas A.} and Rita Bortell and Yocum, {Sue A.} and Smock, {Steven L.} and Meng Zhang and Cory Abate and Victoria Shalhoub and Neil Aronin and Wright, {Kenneth L.} and {van Wijnen}, {Andre J} and Stein, {Janet L.} and Tom Curran and Lian, {Jane B.} and Stein, {Gary S.}",
year = "1990",
language = "English (US)",
volume = "87",
pages = "9990--9994",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "24",

}

TY - JOUR

T1 - Coordinate occupancy of AP-1 sites in the vitamin D-responsive and CCAAT box elements by Fos-Jun in the osteocalcin gene

T2 - Model for phenotype suppression of transcription

AU - Owen, Thomas A.

AU - Bortell, Rita

AU - Yocum, Sue A.

AU - Smock, Steven L.

AU - Zhang, Meng

AU - Abate, Cory

AU - Shalhoub, Victoria

AU - Aronin, Neil

AU - Wright, Kenneth L.

AU - van Wijnen, Andre J

AU - Stein, Janet L.

AU - Curran, Tom

AU - Lian, Jane B.

AU - Stein, Gary S.

PY - 1990

Y1 - 1990

N2 - Osteocalcin, a bone-specific protein and marker of the mature osteoblast, is expressed only in nonproliferating osteoblasts in a mineralizing extracellular matrix, while type I collagen is expressed in proliferating cells. The nuclear proteins encoded by the c-fos and c-jun protooncogenes are expressed during the proliferation period of osteoblast phenotype development. We present evidence that AP-1 (HeLa cell-activating protein 1) sites residing within two promoter elements of the osteocalcin gene bind the Fos-Jun protein complex: the osteocalcin box (OC box; nucleotides -99 to -76), which contains a CCAAT motif as a central element and influences tissue-specific basal levels of osteocalcin gene transcription, and the vitamin D-responsive element (VDRE; nucleotides -462 to -440), which mediates enhancement of osteocalcin gene transcription. Gel electrophoretic mobility-shift analysis demonstrated high AP-1 binding activity in proliferating osteoblasts and dramatic changes in this activity after the down-regulation of proliferation and the initiation of extracellular-matrix mineralization in primary cultures of normal diploid osteoblasts. Methylation interference analysis established at single nucleotide resolution that purified recombinant Fos and Jun proteins bind in a sequence-specific manner to the AP-1 sites within the VDRE and OC box. Similarly, an AP-1 motif within a putative VDRE of the alkaline phosphatase gene, which is also expressed after the completion of proliferation, binds the Fos-Jun complex. These results support a model in which coordinate occupancy of the AP-1 sites in the VDRE and OC box in proliferating osteoblasts may suppress both basal level and vitamin D-enhanced osteocalcin gene transcription as well as transcription of other genes associated with osteoblast differentiation - a phenomenon we describe as phenotype suppression. This model is further supported by binding of the Fos-Jun complex at an AP-1 site in the type αI collagen promoter that is contiguous with, but not overlapping, the VDRE. Such a sequence organization in the collagen VDRE motif is compatible with vitamin D modulation of collagen but not with osteocalcin and alkaline phosphatase expression in proliferating osteoblasts.

AB - Osteocalcin, a bone-specific protein and marker of the mature osteoblast, is expressed only in nonproliferating osteoblasts in a mineralizing extracellular matrix, while type I collagen is expressed in proliferating cells. The nuclear proteins encoded by the c-fos and c-jun protooncogenes are expressed during the proliferation period of osteoblast phenotype development. We present evidence that AP-1 (HeLa cell-activating protein 1) sites residing within two promoter elements of the osteocalcin gene bind the Fos-Jun protein complex: the osteocalcin box (OC box; nucleotides -99 to -76), which contains a CCAAT motif as a central element and influences tissue-specific basal levels of osteocalcin gene transcription, and the vitamin D-responsive element (VDRE; nucleotides -462 to -440), which mediates enhancement of osteocalcin gene transcription. Gel electrophoretic mobility-shift analysis demonstrated high AP-1 binding activity in proliferating osteoblasts and dramatic changes in this activity after the down-regulation of proliferation and the initiation of extracellular-matrix mineralization in primary cultures of normal diploid osteoblasts. Methylation interference analysis established at single nucleotide resolution that purified recombinant Fos and Jun proteins bind in a sequence-specific manner to the AP-1 sites within the VDRE and OC box. Similarly, an AP-1 motif within a putative VDRE of the alkaline phosphatase gene, which is also expressed after the completion of proliferation, binds the Fos-Jun complex. These results support a model in which coordinate occupancy of the AP-1 sites in the VDRE and OC box in proliferating osteoblasts may suppress both basal level and vitamin D-enhanced osteocalcin gene transcription as well as transcription of other genes associated with osteoblast differentiation - a phenomenon we describe as phenotype suppression. This model is further supported by binding of the Fos-Jun complex at an AP-1 site in the type αI collagen promoter that is contiguous with, but not overlapping, the VDRE. Such a sequence organization in the collagen VDRE motif is compatible with vitamin D modulation of collagen but not with osteocalcin and alkaline phosphatase expression in proliferating osteoblasts.

KW - Collagen

KW - Differentiation

KW - Growth control

KW - Oncogenes

KW - Proliferation

KW - Transcription

UR - http://www.scopus.com/inward/record.url?scp=0025599438&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025599438&partnerID=8YFLogxK

M3 - Article

C2 - 2124710

AN - SCOPUS:0025599438

VL - 87

SP - 9990

EP - 9994

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 24

ER -