Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1997 Mar;17(3):1037–1048. doi: 10.1128/mcb.17.3.1037

A minimal regulatory region maintains constitutive expression of the max gene.

M A Peters 1, K G Sollenberger 1, T L Kao 1, E J Taparowsky 1
PMCID: PMC231828  PMID: 9032230

Abstract

Max is a basic helix-loop-helix/leucine zipper protein that forms heterodimers with the Myc family of proteins to promote cell growth and with the Mad/Mxi1 family of proteins to inhibit cell growth. The role of Max as the obligate binding partner for these two protein families necessitates the observed constitutive expression and relatively long half-life of the max mRNA under a variety of growth conditions. In this study, we have used the chicken max gene to map DNA elements maintaining max gene expression in vertebrate cells. We have identified a minimal regulatory region (MRR) that resides within 115 bp of the max translation initiation site and that possesses an overall structure typical of TATA-less promoters. Within the MRR are two consensus binding sites for Sp1, a ubiquitously expressed transcription factor that plays a role in the expression of many constitutive genes. Interestingly, we show that direct binding by Sp1 to these sites is not required for MRR-mediated transcription. Instead, the integrity of a 20-bp DNA element in the MRR is required for transcriptional activity, as is the interaction of this DNA element with a 90-kDa cellular protein. Our data suggest that it is the persistence of this 90-kDa protein in vertebrate cells which drives max gene expression, insulates the max promoter from the dramatic changes in transcription that accompany cell growth and development, and ensures that adequate levels of Max will be available to facilitate the function of the Myc, Mad, and Mxi1 families of proteins.

Full Text

The Full Text of this article is available as a PDF (1.8 MB).

Selected References

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

  1. Amati B., Brooks M. W., Levy N., Littlewood T. D., Evan G. I., Land H. Oncogenic activity of the c-Myc protein requires dimerization with Max. Cell. 1993 Jan 29;72(2):233–245. doi: 10.1016/0092-8674(93)90663-b. [DOI] [PubMed] [Google Scholar]
  2. Amati B., Dalton S., Brooks M. W., Littlewood T. D., Evan G. I., Land H. Transcriptional activation by the human c-Myc oncoprotein in yeast requires interaction with Max. Nature. 1992 Oct 1;359(6394):423–426. doi: 10.1038/359423a0. [DOI] [PubMed] [Google Scholar]
  3. Amati B., Littlewood T. D., Evan G. I., Land H. The c-Myc protein induces cell cycle progression and apoptosis through dimerization with Max. EMBO J. 1993 Dec 15;12(13):5083–5087. doi: 10.1002/j.1460-2075.1993.tb06202.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Amin C., Wagner A. J., Hay N. Sequence-specific transcriptional activation by Myc and repression by Max. Mol Cell Biol. 1993 Jan;13(1):383–390. doi: 10.1128/mcb.13.1.383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ayer D. E., Eisenman R. N. A switch from Myc:Max to Mad:Max heterocomplexes accompanies monocyte/macrophage differentiation. Genes Dev. 1993 Nov;7(11):2110–2119. doi: 10.1101/gad.7.11.2110. [DOI] [PubMed] [Google Scholar]
  6. Ayer D. E., Kretzner L., Eisenman R. N. Mad: a heterodimeric partner for Max that antagonizes Myc transcriptional activity. Cell. 1993 Jan 29;72(2):211–222. doi: 10.1016/0092-8674(93)90661-9. [DOI] [PubMed] [Google Scholar]
  7. Azizkhan J. C., Jensen D. E., Pierce A. J., Wade M. Transcription from TATA-less promoters: dihydrofolate reductase as a model. Crit Rev Eukaryot Gene Expr. 1993;3(4):229–254. [PubMed] [Google Scholar]
  8. Beguinot L., Yamazaki H., Pastan I., Johnson A. C. Biochemical characterization of human GCF transcription factor in tumor cells. Cell Growth Differ. 1995 Jun;6(6):699–706. [PubMed] [Google Scholar]
  9. Berberich S. J., Cole M. D. Casein kinase II inhibits the DNA-binding activity of Max homodimers but not Myc/Max heterodimers. Genes Dev. 1992 Feb;6(2):166–176. doi: 10.1101/gad.6.2.166. [DOI] [PubMed] [Google Scholar]
  10. Berberich S., Hyde-DeRuyscher N., Espenshade P., Cole M. max encodes a sequence-specific DNA-binding protein and is not regulated by serum growth factors. Oncogene. 1992 Apr;7(4):775–779. [PubMed] [Google Scholar]
  11. Blackwood E. M., Eisenman R. N. Max: a helix-loop-helix zipper protein that forms a sequence-specific DNA-binding complex with Myc. Science. 1991 Mar 8;251(4998):1211–1217. doi: 10.1126/science.2006410. [DOI] [PubMed] [Google Scholar]
  12. Blackwood E. M., Lüscher B., Eisenman R. N. Myc and Max associate in vivo. Genes Dev. 1992 Jan;6(1):71–80. doi: 10.1101/gad.6.1.71. [DOI] [PubMed] [Google Scholar]
  13. Boam D. S., Davidson I., Chambon P. A TATA-less promoter containing binding sites for ubiquitous transcription factors mediates cell type-specific regulation of the gene for transcription enhancer factor-1 (TEF-1). J Biol Chem. 1995 Aug 18;270(33):19487–19494. doi: 10.1074/jbc.270.33.19487. [DOI] [PubMed] [Google Scholar]
  14. Coppola J. A., Cole M. D. Constitutive c-myc oncogene expression blocks mouse erythroleukaemia cell differentiation but not commitment. Nature. 1986 Apr 24;320(6064):760–763. doi: 10.1038/320760a0. [DOI] [PubMed] [Google Scholar]
  15. Cornwell M. M., Smith D. E. SP1 activates the MDR1 promoter through one of two distinct G-rich regions that modulate promoter activity. J Biol Chem. 1993 Sep 15;268(26):19505–19511. [PubMed] [Google Scholar]
  16. Courey A. J., Tjian R. Analysis of Sp1 in vivo reveals multiple transcriptional domains, including a novel glutamine-rich activation motif. Cell. 1988 Dec 2;55(5):887–898. doi: 10.1016/0092-8674(88)90144-4. [DOI] [PubMed] [Google Scholar]
  17. Davenport E. A., Taparowsky E. J. Basal levels of max are sufficient for the cotransformation of C3H10T1/2 cells by ras and myc. Exp Cell Res. 1992 Oct;202(2):532–540. doi: 10.1016/0014-4827(92)90108-k. [DOI] [PubMed] [Google Scholar]
  18. Davis A. C., Wims M., Spotts G. D., Hann S. R., Bradley A. A null c-myc mutation causes lethality before 10.5 days of gestation in homozygotes and reduced fertility in heterozygous female mice. Genes Dev. 1993 Apr;7(4):671–682. doi: 10.1101/gad.7.4.671. [DOI] [PubMed] [Google Scholar]
  19. Delgado M. D., Lerga A., Cañelles M., Gómez-Casares M. T., León J. Differential regulation of Max and role of c-Myc during erythroid and myelomonocytic differentiation of K562 cells. Oncogene. 1995 Apr 20;10(8):1659–1665. [PubMed] [Google Scholar]
  20. Dorsey M. J., Tae H. J., Sollenberger K. G., Mascarenhas N. T., Johansen L. M., Taparowsky E. J. B-ATF: a novel human bZIP protein that associates with members of the AP-1 transcription factor family. Oncogene. 1995 Dec 7;11(11):2255–2265. [PubMed] [Google Scholar]
  21. 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]
  22. Farina S. F., Huff J. L., Parsons J. T. Mutations within the 5' half of the avian retrovirus MC29 v-myc gene alter or abolish transformation of chicken embryo fibroblasts and macrophages. J Virol. 1992 May;66(5):2698–2708. doi: 10.1128/jvi.66.5.2698-2708.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Freytag S. O., Dang C. V., Lee W. M. Definition of the activities and properties of c-myc required to inhibit cell differentiation. Cell Growth Differ. 1990 Jul;1(7):339–343. [PubMed] [Google Scholar]
  24. Gallagher R. C., Neil J. C., Fulton R. Cloning and sequence of the feline max, and max 9 transcripts. DNA Seq. 1995;5(5):269–271. doi: 10.3109/10425179509030979. [DOI] [PubMed] [Google Scholar]
  25. Gaubatz S., Meichle A., Eilers M. An E-box element localized in the first intron mediates regulation of the prothymosin alpha gene by c-myc. Mol Cell Biol. 1994 Jun;14(6):3853–3862. doi: 10.1128/mcb.14.6.3853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Geng Y., Johnson L. F. Lack of an initiator element is responsible for multiple transcriptional initiation sites of the TATA-less mouse thymidylate synthase promoter. Mol Cell Biol. 1993 Aug;13(8):4894–4903. doi: 10.1128/mcb.13.8.4894. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Gilladoga A. D., Edelhoff S., Blackwood E. M., Eisenman R. N., Disteche C. M. Mapping of MAX to human chromosome 14 and mouse chromosome 12 by in situ hybridization. Oncogene. 1992 Jun;7(6):1249–1251. [PubMed] [Google Scholar]
  28. 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]
  29. Grayson J., Williams R. S., Yu Y. T., Bassel-Duby R. Synergistic interactions between heterologous upstream activation elements and specific TATA sequences in a muscle-specific promoter. Mol Cell Biol. 1995 Apr;15(4):1870–1878. doi: 10.1128/mcb.15.4.1870. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Gu W., Cechova K., Tassi V., Dalla-Favera R. Opposite regulation of gene transcription and cell proliferation by c-Myc and Max. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2935–2939. doi: 10.1073/pnas.90.7.2935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Haun R. S., Moss J., Vaughan M. Characterization of the human ADP-ribosylation factor 3 promoter. Transcriptional regulation of a TATA-less promoter. J Biol Chem. 1993 Apr 25;268(12):8793–8800. [PubMed] [Google Scholar]
  32. Henriksson M., Lüscher B. Proteins of the Myc network: essential regulators of cell growth and differentiation. Adv Cancer Res. 1996;68:109–182. doi: 10.1016/s0065-230x(08)60353-x. [DOI] [PubMed] [Google Scholar]
  33. Huang H. C., Sundseth R., Hansen U. Transcription factor LSF binds two variant bipartite sites within the SV40 late promoter. Genes Dev. 1990 Feb;4(2):287–298. doi: 10.1101/gad.4.2.287. [DOI] [PubMed] [Google Scholar]
  34. Hurlin P. J., Quéva C., Koskinen P. J., Steingrímsson E., Ayer D. E., Copeland N. G., Jenkins N. A., Eisenman R. N. Mad3 and Mad4: novel Max-interacting transcriptional repressors that suppress c-myc dependent transformation and are expressed during neural and epidermal differentiation. EMBO J. 1995 Nov 15;14(22):5646–5659. doi: 10.1002/j.1460-2075.1995.tb00252.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Izawa M. Molecular cloning and sequencing of rat Max cDNA: castration-induced expression of the 2 kb transcript in male accessory sex organs of rats. Biochim Biophys Acta. 1993 Dec 14;1216(3):492–494. doi: 10.1016/0167-4781(93)90021-5. [DOI] [PubMed] [Google Scholar]
  36. Jaynes J. B., Chamberlain J. S., Buskin J. N., Johnson J. E., Hauschka S. D. Transcriptional regulation of the muscle creatine kinase gene and regulated expression in transfected mouse myoblasts. Mol Cell Biol. 1986 Aug;6(8):2855–2864. doi: 10.1128/mcb.6.8.2855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Jolliff K., Li Y., Johnson L. F. Multiple protein-DNA interactions in the TATAA-less mouse thymidylate synthase promoter. Nucleic Acids Res. 1991 May 11;19(9):2267–2274. doi: 10.1093/nar/19.9.2267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Kadonaga J. T., Carner K. R., Masiarz F. R., Tjian R. Isolation of cDNA encoding transcription factor Sp1 and functional analysis of the DNA binding domain. Cell. 1987 Dec 24;51(6):1079–1090. doi: 10.1016/0092-8674(87)90594-0. [DOI] [PubMed] [Google Scholar]
  39. Kageyama R., Pastan I. Molecular cloning and characterization of a human DNA binding factor that represses transcription. Cell. 1989 Dec 1;59(5):815–825. doi: 10.1016/0092-8674(89)90605-3. [DOI] [PubMed] [Google Scholar]
  40. Kato G. J., Lee W. M., Chen L. L., Dang C. V. Max: functional domains and interaction with c-Myc. Genes Dev. 1992 Jan;6(1):81–92. doi: 10.1101/gad.6.1.81. [DOI] [PubMed] [Google Scholar]
  41. Kelly K., Cochran B. H., Stiles C. D., Leder P. Cell-specific regulation of the c-myc gene by lymphocyte mitogens and platelet-derived growth factor. Cell. 1983 Dec;35(3 Pt 2):603–610. doi: 10.1016/0092-8674(83)90092-2. [DOI] [PubMed] [Google Scholar]
  42. King M. W., Blackwood E. M., Eisenman R. N. Expression of two distinct homologues of Xenopus Max during early development. Cell Growth Differ. 1993 Feb;4(2):85–92. [PubMed] [Google Scholar]
  43. Kitadai Y., Yamazaki H., Yasui W., Kyo E., Yokozaki H., Kajiyama G., Johnson A. C., Pastan I., Tahara E. GC factor represses transcription of several growth factor/receptor genes and causes growth inhibition of human gastric carcinoma cell lines. Cell Growth Differ. 1993 Apr;4(4):291–296. [PubMed] [Google Scholar]
  44. Larsson L. G., Pettersson M., Oberg F., Nilsson K., Lüscher B. Expression of mad, mxi1, max and c-myc during induced differentiation of hematopoietic cells: opposite regulation of mad and c-myc. Oncogene. 1994 Apr;9(4):1247–1252. [PubMed] [Google Scholar]
  45. Lassar A. B., Davis R. L., Wright W. E., Kadesch T., Murre C., Voronova A., Baltimore D., Weintraub H. Functional activity of myogenic HLH proteins requires hetero-oligomerization with E12/E47-like proteins in vivo. Cell. 1991 Jul 26;66(2):305–315. doi: 10.1016/0092-8674(91)90620-e. [DOI] [PubMed] [Google Scholar]
  46. Lewis C. D., Clark S. P., Felsenfeld G., Gould H. An erythrocyte-specific protein that binds to the poly(dG) region of the chicken beta-globin gene promoter. Genes Dev. 1988 Jul;2(7):863–873. doi: 10.1101/gad.2.7.863. [DOI] [PubMed] [Google Scholar]
  47. Lillie J. W., Green M. R. Transcription activation by the adenovirus E1a protein. Nature. 1989 Mar 2;338(6210):39–44. doi: 10.1038/338039a0. [DOI] [PubMed] [Google Scholar]
  48. Littlewood T. D., Amati B., Land H., Evan G. I. Max and c-Myc/Max DNA-binding activities in cell extracts. Oncogene. 1992 Sep;7(9):1783–1792. [PubMed] [Google Scholar]
  49. Lu J., Lee W., Jiang C., Keller E. B. Start site selection by Sp1 in the TATA-less human Ha-ras promoter. J Biol Chem. 1994 Feb 18;269(7):5391–5402. [PubMed] [Google Scholar]
  50. Min S., Mascarenhas N. T., Taparowsky E. J. Functional analysis of the carboxy-terminal transforming region of v-Myc: binding to Max is necessary, but not sufficient, for cellular transformation. Oncogene. 1993 Oct;8(10):2691–2701. [PubMed] [Google Scholar]
  51. Min S., Taparowsky E. J. v-Myc, but not Max, possesses domains that function in both transcription activation and cellular transformation. Oncogene. 1992 Aug;7(8):1531–1540. [PubMed] [Google Scholar]
  52. Miner J. H., Wold B. J. c-myc inhibition of MyoD and myogenin-initiated myogenic differentiation. Mol Cell Biol. 1991 May;11(5):2842–2851. doi: 10.1128/mcb.11.5.2842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Mukherjee B., Morgenbesser S. D., DePinho R. A. Myc family oncoproteins function through a common pathway to transform normal cells in culture: cross-interference by Max and trans-acting dominant mutants. Genes Dev. 1992 Aug;6(8):1480–1492. doi: 10.1101/gad.6.8.1480. [DOI] [PubMed] [Google Scholar]
  54. Mäkelä T. P., Koskinen P. J., Västrik I., Alitalo K. Alternative forms of Max as enhancers or suppressors of Myc-ras cotransformation. Science. 1992 Apr 17;256(5055):373–377. doi: 10.1126/science.256.5055.373. [DOI] [PubMed] [Google Scholar]
  55. Peña A., Reddy C. D., Wu S., Hickok N. J., Reddy E. P., Yumet G., Soprano D. R., Soprano K. J. Regulation of human ornithine decarboxylase expression by the c-Myc.Max protein complex. J Biol Chem. 1993 Dec 25;268(36):27277–27285. [PubMed] [Google Scholar]
  56. Prendergast G. C., Hopewell R., Gorham B. J., Ziff E. B. Biphasic effect of Max on Myc cotransformation activity and dependence on amino- and carboxy-terminal Max functions. Genes Dev. 1992 Dec;6(12A):2429–2439. doi: 10.1101/gad.6.12a.2429. [DOI] [PubMed] [Google Scholar]
  57. Prendergast G. C., Lawe D., Ziff E. B. Association of Myn, the murine homolog of max, with c-Myc stimulates methylation-sensitive DNA binding and ras cotransformation. Cell. 1991 May 3;65(3):395–407. doi: 10.1016/0092-8674(91)90457-a. [DOI] [PubMed] [Google Scholar]
  58. Reddy C. D., Dasgupta P., Saikumar P., Dudek H., Rauscher F. J., 3rd, Reddy E. P. Mutational analysis of Max: role of basic, helix-loop-helix/leucine zipper domains in DNA binding, dimerization and regulation of Myc-mediated transcriptional activation. Oncogene. 1992 Oct;7(10):2085–2092. [PubMed] [Google Scholar]
  59. Schneider I. Cell lines derived from late embryonic stages of Drosophila melanogaster. J Embryol Exp Morphol. 1972 Apr;27(2):353–365. [PubMed] [Google Scholar]
  60. Schreiber-Agus N., Horner J., Torres R., Chiu F. C., DePinho R. A. Zebra fish myc family and max genes: differential expression and oncogenic activity throughout vertebrate evolution. Mol Cell Biol. 1993 May;13(5):2765–2775. doi: 10.1128/mcb.13.5.2765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Sollenberger K. G., Kao T. L., Taparowsky E. J. Structural analysis of the chicken max gene. Oncogene. 1994 Feb;9(2):661–664. [PubMed] [Google Scholar]
  62. Stanton B. R., Perkins A. S., Tessarollo L., Sassoon D. A., Parada L. F. Loss of N-myc function results in embryonic lethality and failure of the epithelial component of the embryo to develop. Genes Dev. 1992 Dec;6(12A):2235–2247. doi: 10.1101/gad.6.12a.2235. [DOI] [PubMed] [Google Scholar]
  63. Taparowsky E. J., Heaney M. L., Parsons J. T. Oncogene-mediated multistep transformation of C3H10T1/2 cells. Cancer Res. 1987 Aug 1;47(15):4125–4129. [PubMed] [Google Scholar]
  64. Västrik I., Koskinen P. J., Alitalo R., Mäkelä T. P. Alternative mRNA forms and open reading frames of the max gene. Oncogene. 1993 Feb;8(2):503–507. [PubMed] [Google Scholar]
  65. Wagner A. J., Le Beau M. M., Diaz M. O., Hay N. Expression, regulation, and chromosomal localization of the Max gene. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):3111–3115. doi: 10.1073/pnas.89.7.3111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. Wang M. M., Reed R. R. Molecular cloning of the olfactory neuronal transcription factor Olf-1 by genetic selection in yeast. Nature. 1993 Jul 8;364(6433):121–126. doi: 10.1038/364121a0. [DOI] [PubMed] [Google Scholar]
  67. Weis L., Reinberg D. Transcription by RNA polymerase II: initiator-directed formation of transcription-competent complexes. FASEB J. 1992 Nov;6(14):3300–3309. doi: 10.1096/fasebj.6.14.1426767. [DOI] [PubMed] [Google Scholar]
  68. Williams T., Admon A., Lüscher B., Tjian R. Cloning and expression of AP-2, a cell-type-specific transcription factor that activates inducible enhancer elements. Genes Dev. 1988 Dec;2(12A):1557–1569. doi: 10.1101/gad.2.12a.1557. [DOI] [PubMed] [Google Scholar]
  69. Yutzey K. E., Kline R. L., Konieczny S. F. An internal regulatory element controls troponin I gene expression. Mol Cell Biol. 1989 Apr;9(4):1397–1405. doi: 10.1128/mcb.9.4.1397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  70. Zervos A. S., Gyuris J., Brent R. Mxi1, a protein that specifically interacts with Max to bind Myc-Max recognition sites. Cell. 1993 Jan 29;72(2):223–232. doi: 10.1016/0092-8674(93)90662-a. [DOI] [PubMed] [Google Scholar]

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

RESOURCES