Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1995 Dec;15(12):7010–7021. doi: 10.1128/mcb.15.12.7010

The human growth hormone gene is regulated by a multicomponent locus control region.

B K Jones 1, B R Monks 1, S A Liebhaber 1, N E Cooke 1
PMCID: PMC230956  PMID: 8524268

Abstract

The five-member human growth hormone (hGH)/chorionic somatomammotropin (hCS) gene cluster encodes the pituitary-specific hGH-N gene and four highly related genes (hGH-V, hCS-A, hCS-B, and hCS-L) that are expressed only in the placenta. When the hGH-N or hCS-A gene, together with all previously identified cis-acting regulatory sequences, was integrated into the mouse genome, it was expressed only sporadically and at low levels in the transgenic target organs. DNase I mapping of chromatin from expressing and nonexpressing cell types was used to identify a pituitary-specific set of DNase I-hypersensitive sites (HS) and a set of HS common to both the pituitary and placenta, centered approximately 15 and 30 kb 5' of hGH-N, respectively. When contained on a cosmid insert in their native genomic configuration, these HS consistently directed high-level, pituitary-specific expression of hGH-N in transgenic mice and appeared to define a locus control region required for hGH-N expression. Individually, each set of HS was able to mediate position-independent hGH-N expression in the pituitary but demonstrated loss of physiologic control and loss of tissue specificity. The gene-proximal set of HS contained a potent enhancer activity in the pituitary, while the more distal set appeared to function primarily to establish site-of-integration independence. These data indicate that synergistic interactions among multiple elements are required to restrict hGH-N transcription to the pituitary and generate appropriate levels of expression. In addition, these results suggest a role for both shared and unique regulatory sequences in locus control region-mediated expression of the hGH/hCS gene cluster in the pituitary and possibly the placenta.

Full Text

The Full Text of this article is available as a PDF (527.2 KB).

Selected References

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

  1. Aliakbar S., Brown P. R., Nicolaides K. H. Localization of CAI and CAII isoenzymes in normal term human placenta by immunofluorescence techniques. Placenta. 1990 Jan-Feb;11(1):35–39. doi: 10.1016/s0143-4004(05)80441-2. [DOI] [PubMed] [Google Scholar]
  2. Aronow B. J., Ebert C. A., Valerius M. T., Potter S. S., Wiginton D. A., Witte D. P., Hutton J. J. Dissecting a locus control region: facilitation of enhancer function by extended enhancer-flanking sequences. Mol Cell Biol. 1995 Feb;15(2):1123–1135. doi: 10.1128/mcb.15.2.1123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barsh G. S., Seeburg P. H., Gelinas R. E. The human growth hormone gene family: structure and evolution of the chromosomal locus. Nucleic Acids Res. 1983 Jun 25;11(12):3939–3958. doi: 10.1093/nar/11.12.3939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bennani-Baiti I. M., Jones B. K., Liebhaber S. A., Cooke N. E. Physical linkage of the human growth hormone gene cluster and the skeletal muscle sodium channel alpha-subunit gene (SCN4A) on chromosome 17. Genomics. 1995 Oct 10;29(3):647–652. doi: 10.1006/geno.1995.9954. [DOI] [PubMed] [Google Scholar]
  5. Birkenmeier E. H., Gordon J. I. Developmental regulation of a gene that encodes a cysteine-rich intestinal protein and maps near the murine immunoglobulin heavy chain locus. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2516–2520. doi: 10.1073/pnas.83.8.2516. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bodner M., Castrillo J. L., Theill L. E., Deerinck T., Ellisman M., Karin M. The pituitary-specific transcription factor GHF-1 is a homeobox-containing protein. Cell. 1988 Nov 4;55(3):505–518. doi: 10.1016/0092-8674(88)90037-2. [DOI] [PubMed] [Google Scholar]
  7. Bonifer C., Hecht A., Saueressig H., Winter D. M., Sippel A. E. Dynamic chromatin: the regulatory domain organization of eukaryotic gene loci. J Cell Biochem. 1991 Oct;47(2):99–108. doi: 10.1002/jcb.240470203. [DOI] [PubMed] [Google Scholar]
  8. Bonifer C., Vidal M., Grosveld F., Sippel A. E. Tissue specific and position independent expression of the complete gene domain for chicken lysozyme in transgenic mice. EMBO J. 1990 Sep;9(9):2843–2848. doi: 10.1002/j.1460-2075.1990.tb07473.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Brem G., Wanke R., Wolf E., Buchmüller T., Müller M., Brenig B., Hermanns W. Multiple consequences of human growth hormone expression in transgenic mice. Mol Biol Med. 1989 Dec;6(6):531–547. [PubMed] [Google Scholar]
  10. Castrillo J. L., Theill L. E., Karin M. Function of the homeodomain protein GHF1 in pituitary cell proliferation. Science. 1991 Jul 12;253(5016):197–199. doi: 10.1126/science.1677216. [DOI] [PubMed] [Google Scholar]
  11. Chen E. Y., Liao Y. C., Smith D. H., Barrera-Saldaña H. A., Gelinas R. E., Seeburg P. H. The human growth hormone locus: nucleotide sequence, biology, and evolution. Genomics. 1989 May;4(4):479–497. doi: 10.1016/0888-7543(89)90271-1. [DOI] [PubMed] [Google Scholar]
  12. Chien C. T., Buck S., Sternglanz R., Shore D. Targeting of SIR1 protein establishes transcriptional silencing at HM loci and telomeres in yeast. Cell. 1993 Nov 5;75(3):531–541. doi: 10.1016/0092-8674(93)90387-6. [DOI] [PubMed] [Google Scholar]
  13. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  14. Choo K. H., Filby G., Greco S., Lau Y. F., Kan Y. W. Cosmid vectors for high efficiency DNA-mediated transformation and gene amplification in mammalian cells: studies with the human growth hormone gene. Gene. 1986;46(2-3):277–286. doi: 10.1016/0378-1119(86)90412-9. [DOI] [PubMed] [Google Scholar]
  15. Chung J. H., Whiteley M., Felsenfeld G. A 5' element of the chicken beta-globin domain serves as an insulator in human erythroid cells and protects against position effect in Drosophila. Cell. 1993 Aug 13;74(3):505–514. doi: 10.1016/0092-8674(93)80052-g. [DOI] [PubMed] [Google Scholar]
  16. Cooke N. E., Liebhaber S. A. Molecular biology of the growth hormone-prolactin gene system. Vitam Horm. 1995;50:385–459. doi: 10.1016/s0083-6729(08)60659-7. [DOI] [PubMed] [Google Scholar]
  17. Dillon N., Grosveld F. Transcriptional regulation of multigene loci: multilevel control. Trends Genet. 1993 Apr;9(4):134–137. doi: 10.1016/0168-9525(93)90208-y. [DOI] [PubMed] [Google Scholar]
  18. Dorer D. R., Henikoff S. Expansions of transgene repeats cause heterochromatin formation and gene silencing in Drosophila. Cell. 1994 Jul 1;77(7):993–1002. doi: 10.1016/0092-8674(94)90439-1. [DOI] [PubMed] [Google Scholar]
  19. Eissenberg J. C., Elgin S. C. Boundary functions in the control of gene expression. Trends Genet. 1991 Oct;7(10):335–340. doi: 10.1016/0168-9525(91)90424-o. [DOI] [PubMed] [Google Scholar]
  20. Engel J. D. Developmental regulation of human beta-globin gene transcription: a switch of loyalties? Trends Genet. 1993 Sep;9(9):304–309. doi: 10.1016/0168-9525(93)90248-g. [DOI] [PubMed] [Google Scholar]
  21. Estes P. A., Cooke N. E., Liebhaber S. A. A native RNA secondary structure controls alternative splice-site selection and generates two human growth hormone isoforms. J Biol Chem. 1992 Jul 25;267(21):14902–14908. [PubMed] [Google Scholar]
  22. Forrester W. C., Epner E., Driscoll M. C., Enver T., Brice M., Papayannopoulou T., Groudine M. A deletion of the human beta-globin locus activation region causes a major alteration in chromatin structure and replication across the entire beta-globin locus. Genes Dev. 1990 Oct;4(10):1637–1649. doi: 10.1101/gad.4.10.1637. [DOI] [PubMed] [Google Scholar]
  23. Forrester W. C., Takegawa S., Papayannopoulou T., Stamatoyannopoulos G., Groudine M. Evidence for a locus activation region: the formation of developmentally stable hypersensitive sites in globin-expressing hybrids. Nucleic Acids Res. 1987 Dec 23;15(24):10159–10177. doi: 10.1093/nar/15.24.10159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Fraser P., Hurst J., Collis P., Grosveld F. DNaseI hypersensitive sites 1, 2 and 3 of the human beta-globin dominant control region direct position-independent expression. Nucleic Acids Res. 1990 Jun 25;18(12):3503–3508. doi: 10.1093/nar/18.12.3503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Fraser P., Pruzina S., Antoniou M., Grosveld F. Each hypersensitive site of the human beta-globin locus control region confers a different developmental pattern of expression on the globin genes. Genes Dev. 1993 Jan;7(1):106–113. doi: 10.1101/gad.7.1.106. [DOI] [PubMed] [Google Scholar]
  26. Gross D. S., Garrard W. T. Nuclease hypersensitive sites in chromatin. Annu Rev Biochem. 1988;57:159–197. doi: 10.1146/annurev.bi.57.070188.001111. [DOI] [PubMed] [Google Scholar]
  27. Grosveld F., van Assendelft G. B., Greaves D. R., Kollias G. Position-independent, high-level expression of the human beta-globin gene in transgenic mice. Cell. 1987 Dec 24;51(6):975–985. doi: 10.1016/0092-8674(87)90584-8. [DOI] [PubMed] [Google Scholar]
  28. Groudine M., Linial M. Chromatin structure and gene expression in germ line and somatic cells. Adv Exp Med Biol. 1986;205:205–243. doi: 10.1007/978-1-4684-5209-9_10. [DOI] [PubMed] [Google Scholar]
  29. Hammer R. E., Palmiter R. D., Brinster R. L. Partial correction of murine hereditary growth disorder by germ-line incorporation of a new gene. Nature. 1984 Sep 6;311(5981):65–67. doi: 10.1038/311065a0. [DOI] [PubMed] [Google Scholar]
  30. Higgs D. R., Wood W. G., Jarman A. P., Sharpe J., Lida J., Pretorius I. M., Ayyub H. A major positive regulatory region located far upstream of the human alpha-globin gene locus. Genes Dev. 1990 Sep;4(9):1588–1601. doi: 10.1101/gad.4.9.1588. [DOI] [PubMed] [Google Scholar]
  31. Huber M. C., Bosch F. X., Sippel A. E., Bonifer C. Chromosomal position effects in chicken lysozyme gene transgenic mice are correlated with suppression of DNase I hypersensitive site formation. Nucleic Acids Res. 1994 Oct 11;22(20):4195–4201. doi: 10.1093/nar/22.20.4195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Imbalzano A. N., Kwon H., Green M. R., Kingston R. E. Facilitated binding of TATA-binding protein to nucleosomal DNA. Nature. 1994 Aug 11;370(6489):481–485. doi: 10.1038/370481a0. [DOI] [PubMed] [Google Scholar]
  33. Ingraham H. A., Chen R. P., Mangalam H. J., Elsholtz H. P., Flynn S. E., Lin C. R., Simmons D. M., Swanson L., Rosenfeld M. G. A tissue-specific transcription factor containing a homeodomain specifies a pituitary phenotype. Cell. 1988 Nov 4;55(3):519–529. doi: 10.1016/0092-8674(88)90038-4. [DOI] [PubMed] [Google Scholar]
  34. Jaenisch R. Transgenic animals. Science. 1988 Jun 10;240(4858):1468–1474. doi: 10.1126/science.3287623. [DOI] [PubMed] [Google Scholar]
  35. Kao T. L., Supowit S. C., Thompson E. A., Meyer W. J., 3rd Immunoreactive growth hormone production by human lymphocyte cell lines. Cell Mol Neurobiol. 1992 Oct;12(5):483–498. doi: 10.1007/BF00711549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Kellum R., Schedl P. A position-effect assay for boundaries of higher order chromosomal domains. Cell. 1991 Mar 8;64(5):941–950. doi: 10.1016/0092-8674(91)90318-s. [DOI] [PubMed] [Google Scholar]
  37. Kennison J. A. Transcriptional activation of Drosophila homeotic genes from distant regulatory elements. Trends Genet. 1993 Mar;9(3):75–79. doi: 10.1016/0168-9525(93)90227-9. [DOI] [PubMed] [Google Scholar]
  38. Lipkin S. M., När A. M., Kalla K. A., Sack R. A., Rosenfeld M. G. Identification of a novel zinc finger protein binding a conserved element critical for Pit-1-dependent growth hormone gene expression. Genes Dev. 1993 Sep;7(9):1674–1687. doi: 10.1101/gad.7.9.1674. [DOI] [PubMed] [Google Scholar]
  39. Loo S., Rine J. Silencers and domains of generalized repression. Science. 1994 Jun 17;264(5166):1768–1771. doi: 10.1126/science.8209257. [DOI] [PubMed] [Google Scholar]
  40. Lytras A., Bock M. E., Dodd J. G., Cattini P. A. Detection of placental growth hormone variant and chorionic somatomammotropin ribonucleic acid expression in human trophoblastic neoplasms by reverse transcriptase-polymerase chain reaction. Endocrinology. 1994 Jun;134(6):2461–2467. doi: 10.1210/endo.134.6.7515000. [DOI] [PubMed] [Google Scholar]
  41. MacLeod J. N., Lee A. K., Liebhaber S. A., Cooke N. E. Developmental control and alternative splicing of the placentally expressed transcripts from the human growth hormone gene cluster. J Biol Chem. 1992 Jul 15;267(20):14219–14226. [PubMed] [Google Scholar]
  42. May G., Enver T. Targeting gene expression to haemopoietic stem cells: a chromatin-dependent upstream element mediates cell type-specific expression of the stem cell antigen CD34. EMBO J. 1995 Feb 1;14(3):564–574. doi: 10.1002/j.1460-2075.1995.tb07032.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Meyuhas O., Perry R. P. Construction and identification of cDNA clones for mouse ribosomal proteins: application for the study of r-protein gene expression. Gene. 1980 Jul;10(2):113–129. doi: 10.1016/0378-1119(80)90129-8. [DOI] [PubMed] [Google Scholar]
  44. Misra-Press A., Cooke N. E., Liebhaber S. A. Complex alternative splicing partially inactivates the human chorionic somatomammotropin-like (hCS-L) gene. J Biol Chem. 1994 Sep 16;269(37):23220–23229. [PubMed] [Google Scholar]
  45. Nachtigal M. W., Nickel B. E., Cattini P. A. Pituitary-specific repression of placental members of the human growth hormone gene family. A possible mechanism for locus regulation. J Biol Chem. 1993 Apr 25;268(12):8473–8479. [PubMed] [Google Scholar]
  46. Nickel B. E., Kardami E., Cattini P. A. Differential expression of human placental growth-hormone variant and chorionic somatomammotropin in culture. Biochem J. 1990 May 1;267(3):653–658. doi: 10.1042/bj2670653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Orkin S. H. Globin gene regulation and switching: circa 1990. Cell. 1990 Nov 16;63(4):665–672. doi: 10.1016/0092-8674(90)90133-y. [DOI] [PubMed] [Google Scholar]
  48. Palmiter R. D., Brinster R. L. Germ-line transformation of mice. Annu Rev Genet. 1986;20:465–499. doi: 10.1146/annurev.ge.20.120186.002341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Peterson C. L., Tamkun J. W. The SWI-SNF complex: a chromatin remodeling machine? Trends Biochem Sci. 1995 Apr;20(4):143–146. doi: 10.1016/s0968-0004(00)88990-2. [DOI] [PubMed] [Google Scholar]
  50. Pruzina S., Hanscombe O., Whyatt D., Grosveld F., Philipsen S. Hypersensitive site 4 of the human beta globin locus control region. Nucleic Acids Res. 1991 Apr 11;19(7):1413–1419. doi: 10.1093/nar/19.7.1413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Rhodes S. J., DiMattia G. E., Rosenfeld M. G. Transcriptional mechanisms in anterior pituitary cell differentiation. Curr Opin Genet Dev. 1994 Oct;4(5):709–717. doi: 10.1016/0959-437x(94)90138-s. [DOI] [PubMed] [Google Scholar]
  52. Shaffer C. D., Wallrath L. L., Elgin S. C. Regulating genes by packaging domains: bits of heterochromatin in euchromatin? Trends Genet. 1993 Feb;9(2):35–37. doi: 10.1016/0168-9525(93)90171-D. [DOI] [PubMed] [Google Scholar]
  53. Stief A., Winter D. M., Strätling W. H., Sippel A. E. A nuclear DNA attachment element mediates elevated and position-independent gene activity. Nature. 1989 Sep 28;341(6240):343–345. doi: 10.1038/341343a0. [DOI] [PubMed] [Google Scholar]
  54. Sun J., Oddoux C., Lazarus A., Gilbert M. T., Catanzaro D. F. Promoter activity of human renin 5'-flanking DNA sequences is activated by the pituitary-specific transcription factor Pit-1. J Biol Chem. 1993 Jan 25;268(3):1505–1508. [PubMed] [Google Scholar]
  55. Svaren J., Chalkley R. The structure and assembly of active chromatin. Trends Genet. 1990 Feb;6(2):52–56. doi: 10.1016/0168-9525(90)90074-g. [DOI] [PubMed] [Google Scholar]
  56. Theill L. E., Karin M. Transcriptional control of GH expression and anterior pituitary development. Endocr Rev. 1993 Dec;14(6):670–689. doi: 10.1210/edrv-14-6-670. [DOI] [PubMed] [Google Scholar]
  57. Townes T. M., Behringer R. R. Human globin locus activation region (LAR): role in temporal control. Trends Genet. 1990 Jul;6(7):219–223. doi: 10.1016/0168-9525(90)90182-6. [DOI] [PubMed] [Google Scholar]
  58. Tsukiyama T., Becker P. B., Wu C. ATP-dependent nucleosome disruption at a heat-shock promoter mediated by binding of GAGA transcription factor. Nature. 1994 Feb 10;367(6463):525–532. doi: 10.1038/367525a0. [DOI] [PubMed] [Google Scholar]
  59. Tuan D., Solomon W., Li Q., London I. M. The "beta-like-globin" gene domain in human erythroid cells. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6384–6388. doi: 10.1073/pnas.82.19.6384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Walker W. H., Fitzpatrick S. L., Saunders G. F. Human placental lactogen transcriptional enhancer. Tissue specificity and binding with specific proteins. J Biol Chem. 1990 Aug 5;265(22):12940–12948. [PubMed] [Google Scholar]
  61. Wallrath L. L., Lu Q., Granok H., Elgin S. C. Architectural variations of inducible eukaryotic promoters: preset and remodeling chromatin structures. Bioessays. 1994 Mar;16(3):165–170. doi: 10.1002/bies.950160306. [DOI] [PubMed] [Google Scholar]
  62. Winston F., Carlson M. Yeast SNF/SWI transcriptional activators and the SPT/SIN chromatin connection. Trends Genet. 1992 Nov;8(11):387–391. doi: 10.1016/0168-9525(92)90300-s. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES