Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1999 May 15;340(Pt 1):69–76.

Glucocorticoid inhibition of human SP-A1 promoter activity in NCI-H441 cells.

R R Hoover 1, K H Thomas 1, J Floros 1
PMCID: PMC1220223  PMID: 10229660

Abstract

Glucocorticoids have complex effects on human surfactant protein (SP) SP-A1 and SP-A2 gene expression that occur at both transcriptional and post-transcriptional levels. In the lung adenocarcinoma cell line NCI-H441, dexamethasone causes a dose-dependent decrease in total SP-A mRNA levels and inhibits SP-A gene transcription. In this study, a deletional analysis of the SP-A1 promoter was performed in order to identify cis-acting elements that mediate dexamethasone responsiveness in NCI-H441 cells. The region -32/+63 relative to the start of SP-A1 transcription mediated both basal promoter activity and dexamethasone repression of transcription. Removal of the region +18/+63 abolished dexamethasone responsiveness, indicating that sequences within this region are necessary for the inhibitory effect. Furthermore, the region -32/+63 formed a sequence-specific DNA-protein complex with NCI-H441 nuclear extract. This DNA-protein complex was induced by dexamethasone exposure and its formation was mediated partially by sequences within the region +26/+63.

Full Text

The Full Text of this article is available as a PDF (191.0 KB).

Selected References

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

  1. Ballard P. L., Hawgood S., Liley H., Wellenstein G., Gonzales L. W., Benson B., Cordell B., White R. T. Regulation of pulmonary surfactant apoprotein SP 28-36 gene in fetal human lung. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9527–9531. doi: 10.1073/pnas.83.24.9527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beato M. Gene regulation by steroid hormones. Cell. 1989 Feb 10;56(3):335–344. doi: 10.1016/0092-8674(89)90237-7. [DOI] [PubMed] [Google Scholar]
  3. Bruns G., Stroh H., Veldman G. M., Latt S. A., Floros J. The 35 kd pulmonary surfactant-associated protein is encoded on chromosome 10. Hum Genet. 1987 May;76(1):58–62. doi: 10.1007/BF00283051. [DOI] [PubMed] [Google Scholar]
  4. Chandran U. R., Attardi B., Friedman R., Zheng Z. w., Roberts J. L., DeFranco D. B. Glucocorticoid repression of the mouse gonadotropin-releasing hormone gene is mediated by promoter elements that are recognized by heteromeric complexes containing glucocorticoid receptor. J Biol Chem. 1996 Aug 23;271(34):20412–20420. doi: 10.1074/jbc.271.34.20412. [DOI] [PubMed] [Google Scholar]
  5. Dekowski S. A., Snyder J. M. The combined effects of insulin and cortisol on surfactant protein mRNA levels. Pediatr Res. 1995 Oct;38(4):513–521. doi: 10.1203/00006450-199510000-00007. [DOI] [PubMed] [Google Scholar]
  6. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Drouin J., Sun Y. L., Chamberland M., Gauthier Y., De Léan A., Nemer M., Schmidt T. J. Novel glucocorticoid receptor complex with DNA element of the hormone-repressed POMC gene. EMBO J. 1993 Jan;12(1):145–156. doi: 10.1002/j.1460-2075.1993.tb05640.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Faisst S., Meyer S. Compilation of vertebrate-encoded transcription factors. Nucleic Acids Res. 1992 Jan 11;20(1):3–26. doi: 10.1093/nar/20.1.3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Floros J., DiAngelo S., Koptides M., Karinch A. M., Rogan P. K., Nielsen H., Spragg R. G., Watterberg K., Deiter G. Human SP-A locus: allele frequencies and linkage disequilibrium between the two surfactant protein A genes. Am J Respir Cell Mol Biol. 1996 Oct;15(4):489–498. doi: 10.1165/ajrcmb.15.4.8879183. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Hoover R. R., Floros J. Organization of the human SP-A and SP-D loci at 10q22-q23. Physical and radiation hybrid mapping reveal gene order and orientation. Am J Respir Cell Mol Biol. 1998 Mar;18(3):353–362. doi: 10.1165/ajrcmb.18.3.3035. [DOI] [PubMed] [Google Scholar]
  12. Iannuzzi D. M., Ertsey R., Ballard P. L. Biphasic glucocorticoid regulation of pulmonary SP-A: characterization of inhibitory process. Am J Physiol. 1993 Mar;264(3 Pt 1):L236–L244. doi: 10.1152/ajplung.1993.264.3.L236. [DOI] [PubMed] [Google Scholar]
  13. Karinch A. M., Deiter G., Ballard P. L., Floros J. Regulation of expression of human SP-A1 and SP-A2 genes in fetal lung explant culture. Biochim Biophys Acta. 1998 Jun 16;1398(2):192–202. doi: 10.1016/s0167-4781(98)00047-5. [DOI] [PubMed] [Google Scholar]
  14. Karinch A. M., Floros J. 5' splicing and allelic variants of the human pulmonary surfactant protein A genes. Am J Respir Cell Mol Biol. 1995 Jan;12(1):77–88. doi: 10.1165/ajrcmb.12.1.7811473. [DOI] [PubMed] [Google Scholar]
  15. Katyal S. L., Singh G., Locker J. Characterization of a second human pulmonary surfactant-associated protein SP-A gene. Am J Respir Cell Mol Biol. 1992 Apr;6(4):446–452. doi: 10.1165/ajrcmb/6.4.446. [DOI] [PubMed] [Google Scholar]
  16. Korfhagen T. R., Glasser S. W., Bruno M. D., McMahan M. J., Whitsett J. A. A portion of the human surfactant protein A (SP-A) gene locus consists of a pseudogene. Am J Respir Cell Mol Biol. 1991 May;4(5):463–469. doi: 10.1165/ajrcmb/4.5.463. [DOI] [PubMed] [Google Scholar]
  17. Kouretas D., Karinch A. M., Rishi A., Melchers K., Floros J. Conservation analysis of rat and human SP-A gene identifies 5' flanking sequences of rat SP-A that bind rat lung nuclear proteins. Exp Lung Res. 1993 Jul-Aug;19(4):485–503. doi: 10.3109/01902149309064359. [DOI] [PubMed] [Google Scholar]
  18. Kumar A. R., Snyder J. M. Differential regulation of SP-A1 and SP-A2 genes by cAMP, glucocorticoids, and insulin. Am J Physiol. 1998 Feb;274(2 Pt 1):L177–L185. doi: 10.1152/ajplung.1998.274.2.L177. [DOI] [PubMed] [Google Scholar]
  19. Kölble K., Lu J., Mole S. E., Kaluz S., Reid K. B. Assignment of the human pulmonary surfactant protein D gene (SFTP4) to 10q22-q23 close to the surfactant protein A gene cluster. Genomics. 1993 Aug;17(2):294–298. doi: 10.1006/geno.1993.1324. [DOI] [PubMed] [Google Scholar]
  20. LeVine A. M., Bruno M. D., Huelsman K. M., Ross G. F., Whitsett J. A., Korfhagen T. R. Surfactant protein A-deficient mice are susceptible to group B streptococcal infection. J Immunol. 1997 May 1;158(9):4336–4340. [PubMed] [Google Scholar]
  21. Li J., Gao E., Mendelson C. R. Cyclic AMP-responsive expression of the surfactant protein-A gene is mediated by increased DNA binding and transcriptional activity of thyroid transcription factor-1. J Biol Chem. 1998 Feb 20;273(8):4592–4600. doi: 10.1074/jbc.273.8.4592. [DOI] [PubMed] [Google Scholar]
  22. Liley H. G., White R. T., Benson B. J., Ballard P. L. Glucocorticoids both stimulate and inhibit production of pulmonary surfactant protein A in fetal human lung. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9096–9100. doi: 10.1073/pnas.85.23.9096. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McCormick S. M., Mendelson C. R. Human SP-A1 and SP-A2 genes are differentially regulated during development and by cAMP and glucocorticoids. Am J Physiol. 1994 Apr;266(4 Pt 1):L367–L374. doi: 10.1152/ajplung.1994.266.4.L367. [DOI] [PubMed] [Google Scholar]
  24. O'Reilly M. A., Gazdar A. F., Morris R. E., Whitsett J. A. Differential effects of glucocorticoid on expression of surfactant proteins in a human lung adenocarcinoma cell line. Biochim Biophys Acta. 1988 Jun 30;970(2):194–204. doi: 10.1016/0167-4889(88)90179-6. [DOI] [PubMed] [Google Scholar]
  25. Odom M. J., Snyder J. M., Boggaram V., Mendelson C. R. Glucocorticoid regulation of the major surfactant associated protein (SP-A) and its messenger ribonucleic acid and of morphological development of human fetal lung in vitro. Endocrinology. 1988 Oct;123(4):1712–1720. doi: 10.1210/endo-123-4-1712. [DOI] [PubMed] [Google Scholar]
  26. Pei L. Identification of a negative glucocorticoid response element in the rat type 1 vasoactive intestinal polypeptide receptor gene. J Biol Chem. 1996 Aug 23;271(34):20879–20884. [PubMed] [Google Scholar]
  27. Planer B. C., Ning Y., Kumar S. A., Ballard P. L. Transcriptional regulation of surfactant proteins SP-A and SP-B by phorbol ester. Biochim Biophys Acta. 1997 Aug 7;1353(2):171–179. doi: 10.1016/s0167-4781(97)00070-5. [DOI] [PubMed] [Google Scholar]
  28. Scavo L. M., Ertsey R., Gao B. Q. Human surfactant proteins A1 and A2 are differentially regulated during development and by soluble factors. Am J Physiol. 1998 Oct;275(4 Pt 1):L653–L669. doi: 10.1152/ajplung.1998.275.4.L653. [DOI] [PubMed] [Google Scholar]
  29. Scheinman R. I., Cogswell P. C., Lofquist A. K., Baldwin A. S., Jr Role of transcriptional activation of I kappa B alpha in mediation of immunosuppression by glucocorticoids. Science. 1995 Oct 13;270(5234):283–286. doi: 10.1126/science.270.5234.283. [DOI] [PubMed] [Google Scholar]
  30. Scheinman R. I., Gualberto A., Jewell C. M., Cidlowski J. A., Baldwin A. S., Jr Characterization of mechanisms involved in transrepression of NF-kappa B by activated glucocorticoid receptors. Mol Cell Biol. 1995 Feb;15(2):943–953. doi: 10.1128/mcb.15.2.943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schüle R., Rangarajan P., Kliewer S., Ransone L. J., Bolado J., Yang N., Verma I. M., Evans R. M. Functional antagonism between oncoprotein c-Jun and the glucocorticoid receptor. Cell. 1990 Sep 21;62(6):1217–1226. doi: 10.1016/0092-8674(90)90397-w. [DOI] [PubMed] [Google Scholar]
  32. White R. T., Damm D., Miller J., Spratt K., Schilling J., Hawgood S., Benson B., Cordell B. Isolation and characterization of the human pulmonary surfactant apoprotein gene. 1985 Sep 26-Oct 2Nature. 317(6035):361–363. doi: 10.1038/317361a0. [DOI] [PubMed] [Google Scholar]
  33. Whitsett J. A., Pilot T., Clark J. C., Weaver T. E. Induction of surfactant protein in fetal lung. Effects of cAMP and dexamethasone on SAP-35 RNA and synthesis. J Biol Chem. 1987 Apr 15;262(11):5256–5261. [PubMed] [Google Scholar]
  34. Young P. P., Mendelson C. R. A CRE-like element plays an essential role in cAMP regulation of human SP-A2 gene in alveolar type II cells. Am J Physiol. 1996 Aug;271(2 Pt 1):L287–L299. doi: 10.1152/ajplung.1996.271.2.L287. [DOI] [PubMed] [Google Scholar]
  35. Young P. P., Mendelson C. R. A GT box element is essential for basal and cyclic adenosine 3',5'-monophosphate regulation of the human surfactant protein A2 gene in alveolar type II cells: evidence for the binding of lung nuclear factors distinct from Sp1. Mol Endocrinol. 1997 Jul;11(8):1082–1093. doi: 10.1210/mend.11.8.9950. [DOI] [PubMed] [Google Scholar]
  36. Zhang G., Zhang L., Duff G. W. A negative regulatory region containing a glucocorticosteroid response element (nGRE) in the human interleukin-1beta gene. DNA Cell Biol. 1997 Feb;16(2):145–152. doi: 10.1089/dna.1997.16.145. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES