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. 1990 Jul;87(13):5129–5133. doi: 10.1073/pnas.87.13.5129

Modifications of protein-DNA interactions in the proximal promoter of a cell-growth-regulated histone gene during onset and progression of osteoblast differentiation.

T A Owen 1, J Holthuis 1, E Markose 1, A J van Wijnen 1, S A Wolfe 1, S R Grimes 1, J B Lian 1, G S Stein 1
PMCID: PMC54275  PMID: 2367528

Abstract

A temporal sequence of interrelated cellular, biochemical, and molecular events which occurs during the progressive expression of the differentiated osteoblast phenotype in primary cultures of fetal rat calvarial cells results in the development of a bone-tissue-like organization. This ordered developmental sequence encompasses three periods: proliferation, matrix maturation, and mineralization. Initially, the cells actively proliferate and synthesize type I collagen. This is followed by a period of matrix organization and maturation and then by a period of extracellular matrix mineralization. At the completion of proliferation, when expression of osteoblast phenotype markers such as alkaline phosphatase is observed, the cell-cycle-related histone genes are down-regulated transcriptionally, suggesting that a key signaling mechanism at this transition point involves modifications of protein-DNA interactions in the regulatory elements of these growth-regulated genes. Our results demonstrate that there is a selective loss of interaction of the promoter binding factor HiNF-D with the site II region of an H4 histone gene proximal promoter that regulates the specificity and level of transcription only when the down-regulation of proliferation is accompanied by modifications in the extracellular matrix that contribute to progression of osteoblast differentiation. Thus, this specific loss of protein-DNA interaction serves as a marker for a key transition point in the osteoblast developmental sequence, where the down-regulation of proliferation is functionally coupled to the appearance of osteoblast phenotypic properties associated with the organization and maturation of an extracellular matrix that becomes competent to mineralize.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aronow M. A., Gerstenfeld L. C., Owen T. A., Tassinari M. S., Stein G. S., Lian J. B. Factors that promote progressive development of the osteoblast phenotype in cultured fetal rat calvaria cells. J Cell Physiol. 1990 May;143(2):213–221. doi: 10.1002/jcp.1041430203. [DOI] [PubMed] [Google Scholar]
  2. Baumbach L. L., Marashi F., Plumb M., Stein G., Stein J. Inhibition of DNA replication coordinately reduces cellular levels of core and H1 histone mRNAs: requirement for protein synthesis. Biochemistry. 1984 Apr 10;23(8):1618–1625. doi: 10.1021/bi00303a006. [DOI] [PubMed] [Google Scholar]
  3. Bellows C. G., Aubin J. E., Heersche J. N., Antosz M. E. Mineralized bone nodules formed in vitro from enzymatically released rat calvaria cell populations. Calcif Tissue Int. 1986 Mar;38(3):143–154. doi: 10.1007/BF02556874. [DOI] [PubMed] [Google Scholar]
  4. Ecarot-Charrier B., Glorieux F. H., van der Rest M., Pereira G. Osteoblasts isolated from mouse calvaria initiate matrix mineralization in culture. J Cell Biol. 1983 Mar;96(3):639–643. doi: 10.1083/jcb.96.3.639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  6. Gerstenfeld L. C., Chipman S. D., Glowacki J., Lian J. B. Expression of differentiated function by mineralizing cultures of chicken osteoblasts. Dev Biol. 1987 Jul;122(1):49–60. doi: 10.1016/0012-1606(87)90331-9. [DOI] [PubMed] [Google Scholar]
  7. Grimes S., Weisz-Carrington P., Daum H., 3rd, Smith J., Green L., Wright K., Stein G., Stein J. A rat histone H4 gene closely associated with the testis-specific H1t gene. Exp Cell Res. 1987 Dec;173(2):534–545. doi: 10.1016/0014-4827(87)90293-x. [DOI] [PubMed] [Google Scholar]
  8. Gundberg C. M., Hauschka P. V., Lian J. B., Gallop P. M. Osteocalcin: isolation, characterization, and detection. Methods Enzymol. 1984;107:516–544. doi: 10.1016/0076-6879(84)07036-1. [DOI] [PubMed] [Google Scholar]
  9. Hanly S. M., Bleecker G. C., Heintz N. Identification of promoter elements necessary for transcriptional regulation of a human histone H4 gene in vitro. Mol Cell Biol. 1985 Feb;5(2):380–389. doi: 10.1128/mcb.5.2.380. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Helms S. R., van Wijnen A. J., Kroeger P., Shiels A., Stewart C., Hirshman J., Stein J. L., Stein G. S. Identification of an enhancer-like element upstream from a cell cycle dependent human H4 histone gene. J Cell Physiol. 1987 Sep;132(3):552–558. doi: 10.1002/jcp.1041320319. [DOI] [PubMed] [Google Scholar]
  11. Holthuis J., Owen T. A., van Wijnen A. J., Wright K. L., Ramsey-Ewing A., Kennedy M. B., Carter R., Cosenza S. C., Soprano K. J., Lian J. B. Tumor cells exhibit deregulation of the cell cycle histone gene promoter factor HiNF-D. Science. 1990 Mar 23;247(4949 Pt 1):1454–1457. doi: 10.1126/science.247.4949.1454. [DOI] [PubMed] [Google Scholar]
  12. Jaffe N. R. Alkaline phosphatase activity, characterization, and subcellular distribution during initial skeletogenesis in the prenatal rat limb. Calcif Tissue Res. 1976 Dec 2;21(3):153–162. doi: 10.1007/BF02547392. [DOI] [PubMed] [Google Scholar]
  13. Kroeger P., Stewart C., Schaap T., van Wijnen A., Hirshman J., Helms S., Stein G., Stein J. Proximal and distal regulatory elements that influence in vivo expression of a cell cycle-dependent human H4 histone gene. Proc Natl Acad Sci U S A. 1987 Jun;84(12):3982–3986. doi: 10.1073/pnas.84.12.3982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lian J., Stewart C., Puchacz E., Mackowiak S., Shalhoub V., Collart D., Zambetti G., Stein G. Structure of the rat osteocalcin gene and regulation of vitamin D-dependent expression. Proc Natl Acad Sci U S A. 1989 Feb;86(4):1143–1147. doi: 10.1073/pnas.86.4.1143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Markose E. R., Stein J. L., Stein G. S., Lian J. B. Vitamin D-mediated modifications in protein-DNA interactions at two promoter elements of the osteocalcin gene. Proc Natl Acad Sci U S A. 1990 Mar;87(5):1701–1705. doi: 10.1073/pnas.87.5.1701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. McGhee J. D., Felsenfeld G. Nucleosome structure. Annu Rev Biochem. 1980;49:1115–1156. doi: 10.1146/annurev.bi.49.070180.005343. [DOI] [PubMed] [Google Scholar]
  17. Meier V. S., Böhni R., Schümperli D. Nucleotide sequence of two mouse histone H4 genes. Nucleic Acids Res. 1989 Jan 25;17(2):795–795. doi: 10.1093/nar/17.2.795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Owen T. A., Aronow M., Shalhoub V., Barone L. M., Wilming L., Tassinari M. S., Kennedy M. B., Pockwinse S., Lian J. B., Stein G. S. Progressive development of the rat osteoblast phenotype in vitro: reciprocal relationships in expression of genes associated with osteoblast proliferation and differentiation during formation of the bone extracellular matrix. J Cell Physiol. 1990 Jun;143(3):420–430. doi: 10.1002/jcp.1041430304. [DOI] [PubMed] [Google Scholar]
  19. Pauli U., Chrysogelos S., Stein G., Stein J., Nick H. Protein-DNA interactions in vivo upstream of a cell cycle-regulated human H4 histone gene. Science. 1987 Jun 5;236(4806):1308–1311. doi: 10.1126/science.3035717. [DOI] [PubMed] [Google Scholar]
  20. Pauli U., Chrysogelos S., Stein J., Stein G. Native genomic blotting: high-resolution mapping of DNase I-hypersensitive sites and protein-DNA interactions. Proc Natl Acad Sci U S A. 1988 Jan;85(1):16–20. doi: 10.1073/pnas.85.1.16. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Plumb M., Stein J., Stein G. Coordinate regulation of multiple histone mRNAs during the cell cycle in HeLa cells. Nucleic Acids Res. 1983 Apr 25;11(8):2391–2410. doi: 10.1093/nar/11.8.2391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Seiler-Tuyns A., Birnstiel M. L. Structure and expression in L-cells of a cloned H4 histone gene of the mouse. J Mol Biol. 1981 Oct 5;151(4):607–625. doi: 10.1016/0022-2836(81)90426-5. [DOI] [PubMed] [Google Scholar]
  23. Seiler-Tuyns A., Paterson B. M. Cell cycle regulation of a mouse histone H4 gene requires the H4 promoter. Mol Cell Biol. 1987 Mar;7(3):1048–1054. doi: 10.1128/mcb.7.3.1048. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Shalhoub V., Gerstenfeld L. C., Collart D., Lian J. B., Stein G. S. Downregulation of cell growth and cell cycle regulated genes during chick osteoblast differentiation with the reciprocal expression of histone gene variants. Biochemistry. 1989 Jun 27;28(13):5318–5322. doi: 10.1021/bi00439a002. [DOI] [PubMed] [Google Scholar]
  25. Sierra F., Stein G., Stein J. Structure and in vitro transcription of a human H4 histone gene. Nucleic Acids Res. 1983 Oct 25;11(20):7069–7086. doi: 10.1093/nar/11.20.7069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Stein G. S., Borun T. W. The synthesis of acidic chromosomal proteins during the cell cycle of HeLa S-3 cells. I. The accelerated accumulation of acidic residual nuclear protein before the initiation of DNA replication. J Cell Biol. 1972 Feb;52(2):292–307. doi: 10.1083/jcb.52.2.292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stein G., Lian J., Stein J., Briggs R., Shalhoub V., Wright K., Pauli U., van Wijnen A. Altered binding of human histone gene transcription factors during the shutdown of proliferation and onset of differentiation in HL-60 cells. Proc Natl Acad Sci U S A. 1989 Mar;86(6):1865–1869. doi: 10.1073/pnas.86.6.1865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Thomas P. S. Hybridization of denatured RNA transferred or dotted nitrocellulose paper. Methods Enzymol. 1983;100:255–266. doi: 10.1016/0076-6879(83)00060-9. [DOI] [PubMed] [Google Scholar]
  29. Tushinski R. J., Sussman P. M., Yu L. Y., Bancroft F. C. Pregrowth hormone messenger RNA: glucocorticoid induction and identification in rat pituitary cells. Proc Natl Acad Sci U S A. 1977 Jun;74(6):2357–2361. doi: 10.1073/pnas.74.6.2357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Weisbrod S. Active chromatin. Nature. 1982 May 27;297(5864):289–295. doi: 10.1038/297289a0. [DOI] [PubMed] [Google Scholar]
  31. Wolfe S. A., Anderson J. V., Grimes S. R., Stein G. S., Stein J. S. Comparison of the structural organization and expression of germinal and somatic rat histone H4 genes. Biochim Biophys Acta. 1989 Mar 1;1007(2):140–150. doi: 10.1016/0167-4781(89)90032-8. [DOI] [PubMed] [Google Scholar]
  32. Yoon K., Buenaga R., Rodan G. A. Tissue specificity and developmental expression of rat osteopontin. Biochem Biophys Res Commun. 1987 Nov 13;148(3):1129–1136. doi: 10.1016/s0006-291x(87)80250-4. [DOI] [PubMed] [Google Scholar]
  33. van Wijnen A. J., Stein J. L., Stein G. S. A nuclear protein with affinity for the 5' flanking region of a cell cycle dependent human H4 histone gene in vitro. Nucleic Acids Res. 1987 Feb 25;15(4):1679–1698. doi: 10.1093/nar/15.4.1679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. van Wijnen A. J., Wright K. L., Lian J. B., Stein J. L., Stein G. S. Human H4 histone gene transcription requires the proliferation-specific nuclear factor HiNF-D. Auxiliary roles for HiNF-C (Sp1-like) and HiNF-A (high mobility group-like). J Biol Chem. 1989 Sep 5;264(25):15034–15042. [PubMed] [Google Scholar]
  35. van Wijnen A. J., Wright K. L., Massung R. F., Gerretsen M., Stein J. L., Stein G. S. Two target sites for protein binding in the promoter region of a cell cycle regulated human H1 histone gene. Nucleic Acids Res. 1988 Jan 25;16(2):571–592. doi: 10.1093/nar/16.2.571. [DOI] [PMC free article] [PubMed] [Google Scholar]

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