Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1993 Nov;12(11):4229–4242. doi: 10.1002/j.1460-2075.1993.tb06107.x

More potent transcriptional activators or a transdominant inhibitor of the HNF1 homeoprotein family are generated by alternative RNA processing.

I Bach 1, M Yaniv 1
PMCID: PMC413717  PMID: 7900999

Abstract

We report the isolation of cDNAs from human liver encoding several isoforms of the hepatocyte nuclear factor homeoproteins HNF1 and vHNF1 generated by the differential use of polyadenylation sites and by alternative splicing. In the novel isoforms intron sequences that are excised in the previously described forms are translated in the same frame as exon sequences until the first termination codon is encountered. Hence, the newly found isoforms all contain different C-terminal domains. For HNF1 it has been shown that its C-terminal region is responsible for the activation of transcription. In transient transfection assays the two novel HNF1 isoforms, HNF1-B and -C, transactivate 5-fold better than the previously described HNF1 protein (HNF1-A). The newly isolated isoform of vHNF1, designated vHNF1-C, is unable to transactivate and behaves as a transdominant repressor when cotransfected with HNF1-A, -B or -C. All of the different isoforms of HNF1 and vHNF1 can form homo- and heterodimers and their mRNAs are differentially expressed in fetal and adult human liver, kidney and intestine, suggesting distinct roles during development. Our studies show that the transactivation domain of the members of the HNF1 homeoprotein family is organized in modules which can be exchanged to generate either more potent transcriptional activators or a transdominant repressor.

Full text

PDF
4236

Images in this article

4231Image
on p.4231
4232Image
on p.4232
4233Image
on p.4233
4234Image
on p.4234
4235Image
on p.4235
4236Image
on p.4236
4238Image
on p.4238

Selected References

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

  1. Bach I., Galcheva-Gargova Z., Mattei M. G., Simon-Chazottes D., Guénet J. L., Cereghini S., Yaniv M. Cloning of human hepatic nuclear factor 1 (HNF1) and chromosomal localization of its gene in man and mouse. Genomics. 1990 Sep;8(1):155–164. doi: 10.1016/0888-7543(90)90238-p. [DOI] [PubMed] [Google Scholar]
  2. Bach I., Mattei M. G., Cereghini S., Yaniv M. Two members of an HNF1 homeoprotein family are expressed in human liver. Nucleic Acids Res. 1991 Jul 11;19(13):3553–3559. doi: 10.1093/nar/19.13.3553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bach I., Pontoglio M., Yaniv M. Structure of the gene encoding hepatocyte nuclear factor 1 (HNF1). Nucleic Acids Res. 1992 Aug 25;20(16):4199–4204. doi: 10.1093/nar/20.16.4199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baumhueter S., Mendel D. B., Conley P. B., Kuo C. J., Turk C., Graves M. K., Edwards C. A., Courtois G., Crabtree G. R. HNF-1 shares three sequence motifs with the POU domain proteins and is identical to LF-B1 and APF. Genes Dev. 1990 Mar;4(3):372–379. doi: 10.1101/gad.4.3.372. [DOI] [PubMed] [Google Scholar]
  5. Blumenfeld M., Maury M., Chouard T., Yaniv M., Condamine H. Hepatic nuclear factor 1 (HNF1) shows a wider distribution than products of its known target genes in developing mouse. Development. 1991 Oct;113(2):589–599. doi: 10.1242/dev.113.2.589. [DOI] [PubMed] [Google Scholar]
  6. Boukamp P., Petrussevska R. T., Breitkreutz D., Hornung J., Markham A., Fusenig N. E. Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line. J Cell Biol. 1988 Mar;106(3):761–771. doi: 10.1083/jcb.106.3.761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burtis K. C., Baker B. S. Drosophila doublesex gene controls somatic sexual differentiation by producing alternatively spliced mRNAs encoding related sex-specific polypeptides. Cell. 1989 Mar 24;56(6):997–1010. doi: 10.1016/0092-8674(89)90633-8. [DOI] [PubMed] [Google Scholar]
  8. Cereghini S., Blumenfeld M., Yaniv M. A liver-specific factor essential for albumin transcription differs between differentiated and dedifferentiated rat hepatoma cells. Genes Dev. 1988 Aug;2(8):957–974. doi: 10.1101/gad.2.8.957. [DOI] [PubMed] [Google Scholar]
  9. Cereghini S., Ott M. O., Power S., Maury M. Expression patterns of vHNF1 and HNF1 homeoproteins in early postimplantation embryos suggest distinct and sequential developmental roles. Development. 1992 Nov;116(3):783–797. doi: 10.1242/dev.116.3.783. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Chouard T., Blumenfeld M., Bach I., Vandekerckhove J., Cereghini S., Yaniv M. A distal dimerization domain is essential for DNA-binding by the atypical HNF1 homeodomain. Nucleic Acids Res. 1990 Oct 11;18(19):5853–5863. doi: 10.1093/nar/18.19.5853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Courtois G., Morgan J. G., Campbell L. A., Fourel G., Crabtree G. R. Interaction of a liver-specific nuclear factor with the fibrinogen and alpha 1-antitrypsin promoters. Science. 1987 Oct 30;238(4827):688–692. doi: 10.1126/science.3499668. [DOI] [PubMed] [Google Scholar]
  13. De Simone V., Cortese R. Transcriptional regulation of liver-specific gene expression. Curr Opin Cell Biol. 1991 Dec;3(6):960–965. doi: 10.1016/0955-0674(91)90114-e. [DOI] [PubMed] [Google Scholar]
  14. De Simone V., De Magistris L., Lazzaro D., Gerstner J., Monaci P., Nicosia A., Cortese R. LFB3, a heterodimer-forming homeoprotein of the LFB1 family, is expressed in specialized epithelia. EMBO J. 1991 Jun;10(6):1435–1443. doi: 10.1002/j.1460-2075.1991.tb07664.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Descombes P., Schibler U. A liver-enriched transcriptional activator protein, LAP, and a transcriptional inhibitory protein, LIP, are translated from the same mRNA. Cell. 1991 Nov 1;67(3):569–579. doi: 10.1016/0092-8674(91)90531-3. [DOI] [PubMed] [Google Scholar]
  16. Dobrazanski P., Noguchi T., Kovary K., Rizzo C. A., Lazo P. S., Bravo R. Both products of the fosB gene, FosB and its short form, FosB/SF, are transcriptional activators in fibroblasts. Mol Cell Biol. 1991 Nov;11(11):5470–5478. doi: 10.1128/mcb.11.11.5470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Early P., Rogers J., Davis M., Calame K., Bond M., Wall R., Hood L. Two mRNAs can be produced from a single immunoglobulin mu gene by alternative RNA processing pathways. Cell. 1980 Jun;20(2):313–319. doi: 10.1016/0092-8674(80)90617-0. [DOI] [PubMed] [Google Scholar]
  18. Emens L. A., Landers D. W., Moss L. G. Hepatocyte nuclear factor 1 alpha is expressed in a hamster insulinoma line and transactivates the rat insulin I gene. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7300–7304. doi: 10.1073/pnas.89.16.7300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Finney M. The homeodomain of the transcription factor LF-B1 has a 21 amino acid loop between helix 2 and helix 3. Cell. 1990 Jan 12;60(1):5–6. doi: 10.1016/0092-8674(90)90708-m. [DOI] [PubMed] [Google Scholar]
  20. Foulkes N. S., Borrelli E., Sassone-Corsi P. CREM gene: use of alternative DNA-binding domains generates multiple antagonists of cAMP-induced transcription. Cell. 1991 Feb 22;64(4):739–749. doi: 10.1016/0092-8674(91)90503-q. [DOI] [PubMed] [Google Scholar]
  21. Foulkes N. S., Mellström B., Benusiglio E., Sassone-Corsi P. Developmental switch of CREM function during spermatogenesis: from antagonist to activator. Nature. 1992 Jan 2;355(6355):80–84. doi: 10.1038/355080a0. [DOI] [PubMed] [Google Scholar]
  22. Foulkes N. S., Sassone-Corsi P. More is better: activators and repressors from the same gene. Cell. 1992 Feb 7;68(3):411–414. doi: 10.1016/0092-8674(92)90178-f. [DOI] [PubMed] [Google Scholar]
  23. Frain M., Swart G., Monaci P., Nicosia A., Stämpfli S., Frank R., Cortese R. The liver-specific transcription factor LF-B1 contains a highly diverged homeobox DNA binding domain. Cell. 1989 Oct 6;59(1):145–157. doi: 10.1016/0092-8674(89)90877-5. [DOI] [PubMed] [Google Scholar]
  24. Gorski K., Carneiro M., Schibler U. Tissue-specific in vitro transcription from the mouse albumin promoter. Cell. 1986 Dec 5;47(5):767–776. doi: 10.1016/0092-8674(86)90519-2. [DOI] [PubMed] [Google Scholar]
  25. Hedley M. L., Maniatis T. Sex-specific splicing and polyadenylation of dsx pre-mRNA requires a sequence that binds specifically to tra-2 protein in vitro. Cell. 1991 May 17;65(4):579–586. doi: 10.1016/0092-8674(91)90090-l. [DOI] [PubMed] [Google Scholar]
  26. Huet C., Sahuquillo-Merino C., Coudrier E., Louvard D. Absorptive and mucus-secreting subclones isolated from a multipotent intestinal cell line (HT-29) provide new models for cell polarity and terminal differentiation. J Cell Biol. 1987 Jul;105(1):345–357. doi: 10.1083/jcb.105.1.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Koenig R. J., Lazar M. A., Hodin R. A., Brent G. A., Larsen P. R., Chin W. W., Moore D. D. Inhibition of thyroid hormone action by a non-hormone binding c-erbA protein generated by alternative mRNA splicing. Nature. 1989 Feb 16;337(6208):659–661. doi: 10.1038/337659a0. [DOI] [PubMed] [Google Scholar]
  28. Kuo C. J., Conley P. B., Hsieh C. L., Francke U., Crabtree G. R. Molecular cloning, functional expression, and chromosomal localization of mouse hepatocyte nuclear factor 1. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9838–9842. doi: 10.1073/pnas.87.24.9838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Lazzaro D., De Simone V., De Magistris L., Lehtonen E., Cortese R. LFB1 and LFB3 homeoproteins are sequentially expressed during kidney development. Development. 1992 Feb;114(2):469–479. doi: 10.1242/dev.114.2.469. [DOI] [PubMed] [Google Scholar]
  30. Leibovitz A., McCombs W. M., 3rd, Johnston D., McCoy C. E., Stinson J. C. New human cancer cell culture lines. I. SW-13, small-cell carcinoma of the adrenal cortex. J Natl Cancer Inst. 1973 Aug;51(2):691–697. [PubMed] [Google Scholar]
  31. Maniatis T. Mechanisms of alternative pre-mRNA splicing. Science. 1991 Jan 4;251(4989):33–34. doi: 10.1126/science.1824726. [DOI] [PubMed] [Google Scholar]
  32. McLauchlan J., Simpson S., Clements J. B. Herpes simplex virus induces a processing factor that stimulates poly(A) site usage. Cell. 1989 Dec 22;59(6):1093–1105. doi: 10.1016/0092-8674(89)90765-4. [DOI] [PubMed] [Google Scholar]
  33. Mendel D. B., Crabtree G. R. HNF-1, a member of a novel class of dimerizing homeodomain proteins. J Biol Chem. 1991 Jan 15;266(2):677–680. [PubMed] [Google Scholar]
  34. Mendel D. B., Hansen L. P., Graves M. K., Conley P. B., Crabtree G. R. HNF-1 alpha and HNF-1 beta (vHNF-1) share dimerization and homeo domains, but not activation domains, and form heterodimers in vitro. Genes Dev. 1991 Jun;5(6):1042–1056. doi: 10.1101/gad.5.6.1042. [DOI] [PubMed] [Google Scholar]
  35. Mendel D. B., Khavari P. A., Conley P. B., Graves M. K., Hansen L. P., Admon A., Crabtree G. R. Characterization of a cofactor that regulates dimerization of a mammalian homeodomain protein. Science. 1991 Dec 20;254(5039):1762–1767. doi: 10.1126/science.1763325. [DOI] [PubMed] [Google Scholar]
  36. Montarras D., Pinset C., Chelly J., Kahn A., Gros F. Expression of MyoD1 coincides with terminal differentiation in determined but inducible muscle cells. EMBO J. 1989 Aug;8(8):2203–2207. doi: 10.1002/j.1460-2075.1989.tb08343.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Nicosia A., Monaci P., Tomei L., De Francesco R., Nuzzo M., Stunnenberg H., Cortese R. A myosin-like dimerization helix and an extra-large homeodomain are essential elements of the tripartite DNA binding structure of LFB1. Cell. 1990 Jun 29;61(7):1225–1236. doi: 10.1016/0092-8674(90)90687-a. [DOI] [PubMed] [Google Scholar]
  38. Olsen J., Laustsen L., Kärnström U., Sjöström H., Norén O. Tissue-specific interactions between nuclear proteins and the aminopeptidase N promoter. J Biol Chem. 1991 Sep 25;266(27):18089–18096. [PubMed] [Google Scholar]
  39. Ott M. O., Rey-Campos J., Cereghini S., Yaniv M. vHNF1 is expressed in epithelial cells of distinct embryonic origin during development and precedes HNF1 expression. Mech Dev. 1991 Dec;36(1-2):47–58. doi: 10.1016/0925-4773(91)90071-d. [DOI] [PubMed] [Google Scholar]
  40. Peterson M. L., Perry R. P. Regulated production of mu m and mu s mRNA requires linkage of the poly(A) addition sites and is dependent on the length of the mu s-mu m intron. Proc Natl Acad Sci U S A. 1986 Dec;83(23):8883–8887. doi: 10.1073/pnas.83.23.8883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Raney A. K., Easton A. J., Milich D. R., McLachlan A. Promoter-specific transactivation of hepatitis B virus transcription by a glutamine- and proline-rich domain of hepatocyte nuclear factor 1. J Virol. 1991 Nov;65(11):5774–5781. doi: 10.1128/jvi.65.11.5774-5781.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Rey-Campos J., Chouard T., Yaniv M., Cereghini S. vHNF1 is a homeoprotein that activates transcription and forms heterodimers with HNF1. EMBO J. 1991 Jun;10(6):1445–1457. doi: 10.1002/j.1460-2075.1991.tb07665.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Roman C., Cohn L., Calame K. A dominant negative form of transcription activator mTFE3 created by differential splicing. Science. 1991 Oct 4;254(5028):94–97. doi: 10.1126/science.1840705. [DOI] [PubMed] [Google Scholar]
  44. Sheets M. D., Ogg S. C., Wickens M. P. Point mutations in AAUAAA and the poly (A) addition site: effects on the accuracy and efficiency of cleavage and polyadenylation in vitro. Nucleic Acids Res. 1990 Oct 11;18(19):5799–5805. doi: 10.1093/nar/18.19.5799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Soule H. D., Vazguez J., Long A., Albert S., Brennan M. A human cell line from a pleural effusion derived from a breast carcinoma. J Natl Cancer Inst. 1973 Nov;51(5):1409–1416. doi: 10.1093/jnci/51.5.1409. [DOI] [PubMed] [Google Scholar]
  46. Sourdive D. J., Chouard T., Yaniv M. The HNF1 C-terminal domain contributes to transcriptional activity and modulates nuclear localisation. C R Acad Sci III. 1993;316(4):385–394. [PubMed] [Google Scholar]
  47. Struhl K. Mechanisms for diversity in gene expression patterns. Neuron. 1991 Aug;7(2):177–181. doi: 10.1016/0896-6273(91)90256-y. [DOI] [PubMed] [Google Scholar]
  48. Tilghman S. M., Belayew A. Transcriptional control of the murine albumin/alpha-fetoprotein locus during development. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5254–5257. doi: 10.1073/pnas.79.17.5254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Tomei L., Cortese R., De Francesco R. A POU-A related region dictates DNA binding specificity of LFB1/HNF1 by orienting the two XL-homeodomains in the dimer. EMBO J. 1992 Nov;11(11):4119–4129. doi: 10.1002/j.1460-2075.1992.tb05505.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Tripodi M., Abbott C., Vivian N., Cortese R., Lovell-Badge R. Disruption of the LF-A1 and LF-B1 binding sites in the human alpha-1-antitrypsin gene has a differential effect during development in transgenic mice. EMBO J. 1991 Nov;10(11):3177–3182. doi: 10.1002/j.1460-2075.1991.tb04879.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Tronche F., Rollier A., Bach I., Weiss M. C., Yaniv M. The rat albumin promoter: cooperation with upstream elements is required when binding of APF/HNF1 to the proximal element is partially impaired by mutation or bacterial methylation. Mol Cell Biol. 1989 Nov;9(11):4759–4766. doi: 10.1128/mcb.9.11.4759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Tronche F., Rollier A., Herbomel P., Bach I., Cereghini S., Weiss M., Yaniv M. Anatomy of the rat albumin promoter. Mol Biol Med. 1990 Apr;7(2):173–185. [PubMed] [Google Scholar]
  54. Tronche F., Yaniv M. HNF1, a homeoprotein member of the hepatic transcription regulatory network. Bioessays. 1992 Sep;14(9):579–587. doi: 10.1002/bies.950140902. [DOI] [PubMed] [Google Scholar]
  55. Tyner A. L., Godbout R., Compton R. S., Tilghman S. M. The ontogeny of alpha-fetoprotein gene expression in the mouse gastrointestinal tract. J Cell Biol. 1990 Apr;110(4):915–927. doi: 10.1083/jcb.110.4.915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Wickens M. How the messenger got its tail: addition of poly(A) in the nucleus. Trends Biochem Sci. 1990 Jul;15(7):277–281. doi: 10.1016/0968-0004(90)90054-f. [DOI] [PubMed] [Google Scholar]
  57. Xanthopoulos K. G., Prezioso V. R., Chen W. S., Sladek F. M., Cortese R., Darnell J. E., Jr The different tissue transcription patterns of genes for HNF-1, C/EBP, HNF-3, and HNF-4, protein factors that govern liver-specific transcription. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3807–3811. doi: 10.1073/pnas.88.9.3807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Yee C., Krishnan-Hewlett I., Baker C. C., Schlegel R., Howley P. M. Presence and expression of human papillomavirus sequences in human cervical carcinoma cell lines. Am J Pathol. 1985 Jun;119(3):361–366. [PMC free article] [PubMed] [Google Scholar]
  59. Yen J., Wisdom R. M., Tratner I., Verma I. M. An alternative spliced form of FosB is a negative regulator of transcriptional activation and transformation by Fos proteins. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5077–5081. doi: 10.1073/pnas.88.12.5077. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES