Abstract
We have analyzed the regulation of the alpha gonadotropin gene in eutopic placental cells and ectopic tumor cells by constructing a series of plasmid vectors containing alpha genomic 5' flanking DNA placed upstream of the gene encoding the bacterial enzyme chloramphenicol acetyltransferase (CAT). These plasmid DNAs were transfected into a eutopic (JAr) and an ectopic (HeLa) cell line. Both cell types expressed the CAT gene from plasmid constructs containing as much as 1,500 base pairs (bp) and as little as 140 bp of alpha 5' flanking DNA; JAr cells were considerably more efficient than HeLa cells. Ectopic and eutopic cells differed qualitatively in their expression from these alpha-CAT constructs when cells were treated with cAMP or butyrate. Butyrate induced alpha expression in HeLa cells but not in JAr cells, while cAMP induced expression in JAr cells. These results are consistent with and extend previous observations suggesting that there are cell-specific differences in the regulation of alpha gene expression in ectopic and eutopic cells. However, by using deletion constructs of the alpha-CAT gene, we found that the basal expression and cell-specific induction of the alpha gene in ectopic and eutopic cells were dependent on the same 140 bp of alpha 5' flanking DNA. These 140 bp were sequenced and found to contain a 9-bp stretch of DNA homologous with the consensus viral enhancer sequence. Such features of alpha expression common to both ectopic and eutopic cells may be involved in the coordinate expression of the alpha gene and the tumorigenic phenotype observed in each cell type.
Full text
PDF










Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Banerji J., Olson L., Schaffner W. A lymphocyte-specific cellular enhancer is located downstream of the joining region in immunoglobulin heavy chain genes. Cell. 1983 Jul;33(3):729–740. doi: 10.1016/0092-8674(83)90015-6. [DOI] [PubMed] [Google Scholar]
- Boothby M., Ruddon R. W., Anderson C., McWilliams D., Boime I. A single gonadotropin alpha-subunit gene in normal tissue and tumor-derived cell lines. J Biol Chem. 1981 May 25;256(10):5121–5127. [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Brickell P. M., Latchman D. S., Murphy D., Willison K., Rigby P. W. Activation of a Qa/Tla class I major histocompatibility antigen gene is a general feature of oncogenesis in the mouse. Nature. 1983 Dec 22;306(5945):756–760. doi: 10.1038/306756a0. [DOI] [PubMed] [Google Scholar]
- Chandler V. L., Maler B. A., Yamamoto K. R. DNA sequences bound specifically by glucocorticoid receptor in vitro render a heterologous promoter hormone responsive in vivo. Cell. 1983 Jun;33(2):489–499. doi: 10.1016/0092-8674(83)90430-0. [DOI] [PubMed] [Google Scholar]
- Charnay P., Treisman R., Mellon P., Chao M., Axel R., Maniatis T. Differences in human alpha- and beta-globin gene expression in mouse erythroleukemia cells: the role of intragenic sequences. Cell. 1984 Aug;38(1):251–263. doi: 10.1016/0092-8674(84)90547-6. [DOI] [PubMed] [Google Scholar]
- Chou J. Y. Establishment of clonal human placental cells synthesizing human choriogonadotropin. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1854–1858. doi: 10.1073/pnas.75.4.1854. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chou J. Y., Robinson J. C., Wang S. S. Effects of sodium butyrate on synthesis of human chorionic gonadotrophin in trophoblastic and non-trophoblastic tumours. Nature. 1977 Aug 11;268(5620):543–544. doi: 10.1038/268543a0. [DOI] [PubMed] [Google Scholar]
- Compton J. G., Schrader W. T., O'Malley B. W. DNA sequence preference of the progesterone receptor. Proc Natl Acad Sci U S A. 1983 Jan;80(1):16–20. doi: 10.1073/pnas.80.1.16. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Darnell R. B. Independent regulation by sodium butyrate of gonadotropin alpha gene expression and cell cycle progression in HeLa cells. Mol Cell Biol. 1984 May;4(5):829–839. doi: 10.1128/mcb.4.5.829. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fiddes J. C., Goodman H. M. The gene encoding the common alpha subunit of the four human glycoprotein hormones. J Mol Appl Genet. 1981;1(1):3–18. [PubMed] [Google Scholar]
- Ghosh N. K., Rukenstein A., Cox R. P. Induction of human choriogonadotropin in heLa-cell cultures by aliphatic monocarboxylates and inhibitors of deoxyribonucleic acid synthesis. Biochem J. 1977 Aug 15;166(2):265–274. doi: 10.1042/bj1660265. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gillies S. D., Morrison S. L., Oi V. T., Tonegawa S. A tissue-specific transcription enhancer element is located in the major intron of a rearranged immunoglobulin heavy chain gene. Cell. 1983 Jul;33(3):717–728. doi: 10.1016/0092-8674(83)90014-4. [DOI] [PubMed] [Google Scholar]
- Goodwin R. G., Moncman C. L., Rottman F. M., Nilson J. H. Characterization and nucleotide sequence of the gene for the common alpha subunit of the bovine pituitary glycoprotein hormones. Nucleic Acids Res. 1983 Oct 11;11(19):6873–6882. doi: 10.1093/nar/11.19.6873. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gorman C. M., Merlino G. T., Willingham M. C., Pastan I., Howard B. H. The Rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6777–6781. doi: 10.1073/pnas.79.22.6777. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Hoshina M., Boothby M., Boime I. Cytological localization of chorionic gonadotropin alpha and placental lactogen mRNAs during development of the human placenta. J Cell Biol. 1982 Apr;93(1):190–198. doi: 10.1083/jcb.93.1.190. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hussa R. O. Biosynthesis of human chorionic gonadotropin. Endocr Rev. 1980 Summer;1(3):268–294. doi: 10.1210/edrv-1-3-268. [DOI] [PubMed] [Google Scholar]
- Hussa R. O., Pattillo R. A., Ruckert A. C., Scheuermann K. W. Effects of butyrate and dibutyryl cyclic AMP on hCG-secreting trophoblastic and non-trophoblastic cells. J Clin Endocrinol Metab. 1978 Jan;46(1):69–76. doi: 10.1210/jcem-46-1-69. [DOI] [PubMed] [Google Scholar]
- Hussa R. O., Story M. T., Pattillo R. A., Kemp R. G. Effect of cyclic 3':5'-AMP derivatives prostaglandins and related agents on human chorionic gonadotropin secretion in human malignant trophoblast in culture. In Vitro. 1977 Jul;13(7):443–449. doi: 10.1007/BF02615105. [DOI] [PubMed] [Google Scholar]
- Imperiale M. J., Hart R. P., Nevins J. R. An enhancer-like element in the adenovirus E2 promoter contains sequences essential for uninduced and E1A-induced transcription. Proc Natl Acad Sci U S A. 1985 Jan;82(2):381–385. doi: 10.1073/pnas.82.2.381. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Karin M., Haslinger A., Holtgreve H., Richards R. I., Krauter P., Westphal H. M., Beato M. Characterization of DNA sequences through which cadmium and glucocorticoid hormones induce human metallothionein-IIA gene. Nature. 1984 Apr 5;308(5959):513–519. doi: 10.1038/308513a0. [DOI] [PubMed] [Google Scholar]
- Keller J. M., Alwine J. C. Activation of the SV40 late promoter: direct effects of T antigen in the absence of viral DNA replication. Cell. 1984 Feb;36(2):381–389. doi: 10.1016/0092-8674(84)90231-9. [DOI] [PubMed] [Google Scholar]
- Khoury G., Gruss P. Enhancer elements. Cell. 1983 Jun;33(2):313–314. doi: 10.1016/0092-8674(83)90410-5. [DOI] [PubMed] [Google Scholar]
- MIDGLEY A. R., PIERCE G. B., Jr, DENEAU G. A., GOSLING J. R. Morphogenesis of syncytiotrophoblast in vivo: an autoradiographic demonstration. Science. 1963 Jul 26;141(3578):349–350. doi: 10.1126/science.141.3578.349. [DOI] [PubMed] [Google Scholar]
- Majors J., Varmus H. E. A small region of the mouse mammary tumor virus long terminal repeat confers glucocorticoid hormone regulation on a linked heterologous gene. Proc Natl Acad Sci U S A. 1983 Oct;80(19):5866–5870. doi: 10.1073/pnas.80.19.5866. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
- PIERCE G. B., Jr, MIDGLEY A. R., Jr THE ORIGIN AND FUNCTION OF HUMAN SYNCYTIOTROPHOBLASTIC GIANT CELLS. Am J Pathol. 1963 Aug;43:153–173. [PMC free article] [PubMed] [Google Scholar]
- Pijnenborg R., Robertson W. B., Brosens I., Dixon G. Review article: trophoblast invasion and the establishment of haemochorial placentation in man and laboratory animals. Placenta. 1981 Jan-Mar;2(1):71–91. doi: 10.1016/s0143-4004(81)80042-2. [DOI] [PubMed] [Google Scholar]
- Queen C., Baltimore D. Immunoglobulin gene transcription is activated by downstream sequence elements. Cell. 1983 Jul;33(3):741–748. doi: 10.1016/0092-8674(83)90016-8. [DOI] [PubMed] [Google Scholar]
- Rosenthal N., Kress M., Gruss P., Khoury G. BK viral enhancer element and a human cellular homolog. Science. 1983 Nov 18;222(4625):749–755. doi: 10.1126/science.6314501. [DOI] [PubMed] [Google Scholar]
- Ruddon R. W., Anderson C., Meade-Cobun K. S. Stimulation of synthesis and secretion of chorionic gonadotropin subunits by eutopic and ectopic hormone-producing human cell lines. Cancer Res. 1980 Dec;40(12):4519–4523. [PubMed] [Google Scholar]
- Ruddon R. W., Anderson C., Meade K. S., Aldenderfer P. H., Neuwald P. D. Content of gonadotropins in cultured human malignant cells and effects of sodium butyrate treatment on gonadotropin secretion by HeLa cells. Cancer Res. 1979 Oct;39(10):3885–3892. [PubMed] [Google Scholar]
- Smith H. O. Recovery of DNA from gels. Methods Enzymol. 1980;65(1):371–380. doi: 10.1016/s0076-6879(80)65048-4. [DOI] [PubMed] [Google Scholar]
- Stanbridge E. J., Der C. J., Doersen C. J., Nishimi R. Y., Peehl D. M., Weissman B. E., Wilkinson J. E. Human cell hybrids: analysis of transformation and tumorigenicity. Science. 1982 Jan 15;215(4530):252–259. doi: 10.1126/science.7053574. [DOI] [PubMed] [Google Scholar]
- Walker M. D., Edlund T., Boulet A. M., Rutter W. J. Cell-specific expression controlled by the 5'-flanking region of insulin and chymotrypsin genes. Nature. 1983 Dec 8;306(5943):557–561. doi: 10.1038/306557a0. [DOI] [PubMed] [Google Scholar]
- Weiher H., König M., Gruss P. Multiple point mutations affecting the simian virus 40 enhancer. Science. 1983 Feb 11;219(4585):626–631. doi: 10.1126/science.6297005. [DOI] [PubMed] [Google Scholar]
- Wigler M., Pellicer A., Silverstein S., Axel R., Urlaub G., Chasin L. DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1373–1376. doi: 10.1073/pnas.76.3.1373. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wright S., Rosenthal A., Flavell R., Grosveld F. DNA sequences required for regulated expression of beta-globin genes in murine erythroleukemia cells. Cell. 1984 Aug;38(1):265–273. doi: 10.1016/0092-8674(84)90548-8. [DOI] [PubMed] [Google Scholar]