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. 1990 Feb 25;18(4):957–961. doi: 10.1093/nar/18.4.957

Analysis of the human liver/bone/kidney alkaline phosphatase promoter in vivo and in vitro.

M Kiledjian 1, T Kadesch 1
PMCID: PMC330350  PMID: 2156238

Abstract

We have carried out an analysis of the promoter for the human liver/bone/kidney alkaline phosphatase (LBK AP) gene. Using transient transfection assays, the intact promoter directs equal expression of a linked cat gene in Saos-2 cells (osteoblast-derived cells which express very high levels of endogenous LBK AP mRNA) and in HeLa and HepG2 cells (which express low levels of endogenous message). The activity of the transfected promoter apparently mimics the true in vivo situation since nuclear run-on assays employing Saos-2 and HeLa cells indicate that the endogenous gene is transcribed at approximately the same rate in these two cell types. Transfections of a series of 5' deletion mutants indicate that promoter activity is dependent on multiple motifs, which possibly include several putative Sp1 binding sites and a TATA box. The LBK AP promoter also directs accurate transcription initiation in HeLa whole cell extracts and in vitro activities of the 5' deletion mutants also suggest that the promoter utilizes multiple motifs.

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Selected References

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

  1. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  2. Buchman A. R., Fromm M., Berg P. Complex regulation of simian virus 40 early-region transcription from different overlapping promoters. Mol Cell Biol. 1984 Sep;4(9):1900–1914. doi: 10.1128/mcb.4.9.1900. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  4. Dynan W. S., Tjian R. Isolation of transcription factors that discriminate between different promoters recognized by RNA polymerase II. Cell. 1983 Mar;32(3):669–680. doi: 10.1016/0092-8674(83)90053-3. [DOI] [PubMed] [Google Scholar]
  5. Dynan W. S., Tjian R. The promoter-specific transcription factor Sp1 binds to upstream sequences in the SV40 early promoter. Cell. 1983 Nov;35(1):79–87. doi: 10.1016/0092-8674(83)90210-6. [DOI] [PubMed] [Google Scholar]
  6. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  8. Harris H. Multilocus enzyme systems and the evolution of gene expression: the alkaline phosphatases as a model example. Harvey Lect. 1980;76:95–124. [PubMed] [Google Scholar]
  9. Herbomel P., Bourachot B., Yaniv M. Two distinct enhancers with different cell specificities coexist in the regulatory region of polyoma. Cell. 1984 Dec;39(3 Pt 2):653–662. doi: 10.1016/0092-8674(84)90472-0. [DOI] [PubMed] [Google Scholar]
  10. Jones N. C., Rigby P. W., Ziff E. B. Trans-acting protein factors and the regulation of eukaryotic transcription: lessons from studies on DNA tumor viruses. Genes Dev. 1988 Mar;2(3):267–281. doi: 10.1101/gad.2.3.267. [DOI] [PubMed] [Google Scholar]
  11. Kadesch T., Berg P. Effects of the position of the simian virus 40 enhancer on expression of multiple transcription units in a single plasmid. Mol Cell Biol. 1986 Jul;6(7):2593–2601. doi: 10.1128/mcb.6.7.2593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kadesch T., Zervos P., Ruezinsky D. Functional analysis of the murine IgH enhancer: evidence for negative control of cell-type specificity. Nucleic Acids Res. 1986 Oct 24;14(20):8209–8221. doi: 10.1093/nar/14.20.8209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Knowles B. B., Howe C. C., Aden D. P. Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. Science. 1980 Jul 25;209(4455):497–499. doi: 10.1126/science.6248960. [DOI] [PubMed] [Google Scholar]
  14. Manley J. L., Fire A., Samuels M., Sharp P. A. In vitro transcription: whole-cell extract. Methods Enzymol. 1983;101:568–582. doi: 10.1016/0076-6879(83)01038-1. [DOI] [PubMed] [Google Scholar]
  15. McKnight S., Tjian R. Transcriptional selectivity of viral genes in mammalian cells. Cell. 1986 Sep 12;46(6):795–805. doi: 10.1016/0092-8674(86)90061-9. [DOI] [PubMed] [Google Scholar]
  16. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rodan S. B., Imai Y., Thiede M. A., Wesolowski G., Thompson D., Bar-Shavit Z., Shull S., Mann K., Rodan G. A. Characterization of a human osteosarcoma cell line (Saos-2) with osteoblastic properties. Cancer Res. 1987 Sep 15;47(18):4961–4966. [PubMed] [Google Scholar]
  18. Weber B., Horiguchi J., Luebbers R., Sherman M., Kufe D. Posttranscriptional stabilization of c-fms mRNA by a labile protein during human monocytic differentiation. Mol Cell Biol. 1989 Feb;9(2):769–775. doi: 10.1128/mcb.9.2.769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Weiss M. J., Ray K., Henthorn P. S., Lamb B., Kadesch T., Harris H. Structure of the human liver/bone/kidney alkaline phosphatase gene. J Biol Chem. 1988 Aug 25;263(24):12002–12010. [PubMed] [Google Scholar]
  20. Wingender E. Compilation of transcription regulating proteins. Nucleic Acids Res. 1988 Mar 25;16(5):1879–1902. doi: 10.1093/nar/16.5.1879. [DOI] [PMC free article] [PubMed] [Google Scholar]

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