Abstract
The ability of vertebrate metallothionein (MT) genes to be induced by heavy metals is controlled by metal regulatory elements (MREs) present in the promoter in multiple, non-identical copies. The binding specificity of the mouse L-cell nuclear factor(s) that interact with the element MREd of the mouse MT-I gene was analyzed by in vitro footprinting, protein blotting, and UV cross-linking assays. In vitro footprinting analyses revealed that synthetic oligodeoxynucleotides (oligomers) corresponding to the metal regulatory elements MREa, MREb, MREc, MREd and MREe of the mouse MT-I gene, as well as the MRE4 of the human MT-IIA gene and the MREa of the trout MT-B gene, all competed for the nuclear protein species binding to the MREd region of the mouse MT-I gene, the MREe oligomer being the weakest competitor. In addition, protein blotting experiments revealed that a nuclear protein of 108 kDa, termed metal element protein-1 (MEP-1), which specifically binds with high affinity to mouse MREd, binds with different affinities to the other mouse MRE elements, mimicking their relative transcriptional strength in vivo: MREd greater than or equal to MREa = MREc greater than MREb greater than MREe greater than MREf. Similarly, human MRE4 and trout MREa bind to MEP-1. A protein similar in size to MEP-1 was also detected in HeLa-cell nuclear extracts. In UV cross-linking experiments the major protein species, complexed with mouse MREd oligomers, migrated on a denaturating gel with an apparent Mr of 115,000 and was detected using each of the mouse MRE oligomers tested. These results show that a mouse nuclear factor can bind to multiple MREs in mouse, trout, and human MT genes.
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Selected References
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- Andersen R. D., Taplitz S. J., Oberbauer A. M., Calame K. L., Herschman H. R. Metal-dependent binding of a nuclear factor to the rat metallothionein-I promoter. Nucleic Acids Res. 1990 Oct 25;18(20):6049–6055. doi: 10.1093/nar/18.20.6049. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Andersen R. D., Taplitz S. J., Wong S., Bristol G., Larkin B., Herschman H. R. Metal-dependent binding of a factor in vivo to the metal-responsive elements of the metallothionein 1 gene promoter. Mol Cell Biol. 1987 Oct;7(10):3574–3581. doi: 10.1128/mcb.7.10.3574. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Buchman C., Skroch P., Welch J., Fogel S., Karin M. The CUP2 gene product, regulator of yeast metallothionein expression, is a copper-activated DNA-binding protein. Mol Cell Biol. 1989 Sep;9(9):4091–4095. doi: 10.1128/mcb.9.9.4091. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Butler G., Thiele D. J. ACE2, an activator of yeast metallothionein expression which is homologous to SWI5. Mol Cell Biol. 1991 Jan;11(1):476–485. doi: 10.1128/mcb.11.1.476. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carter A. D., Felber B. K., Walling M. J., Jubier M. F., Schmidt C. J., Hamer D. H. Duplicated heavy metal control sequences of the mouse metallothionein-I gene. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7392–7396. doi: 10.1073/pnas.81.23.7392. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carthew R. W., Chodosh L. A., Sharp P. A. The major late transcription factor binds to and activates the mouse metallothionein I promoter. Genes Dev. 1987 Nov;1(9):973–980. doi: 10.1101/gad.1.9.973. [DOI] [PubMed] [Google Scholar]
- Chavrier P., Vesque C., Galliot B., Vigneron M., Dollé P., Duboule D., Charnay P. The segment-specific gene Krox-20 encodes a transcription factor with binding sites in the promoter region of the Hox-1.4 gene. EMBO J. 1990 Apr;9(4):1209–1218. doi: 10.1002/j.1460-2075.1990.tb08228.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Culotta V. C., Hamer D. H. Fine mapping of a mouse metallothionein gene metal response element. Mol Cell Biol. 1989 Mar;9(3):1376–1380. doi: 10.1128/mcb.9.3.1376. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Culotta V. C., Hsu T., Hu S., Fürst P., Hamer D. Copper and the ACE1 regulatory protein reversibly induce yeast metallothionein gene transcription in a mouse extract. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8377–8381. doi: 10.1073/pnas.86.21.8377. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fürst P., Hamer D. Cooperative activation of a eukaryotic transcription factor: interaction between Cu(I) and yeast ACE1 protein. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5267–5271. doi: 10.1073/pnas.86.14.5267. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fürst P., Hu S., Hackett R., Hamer D. Copper activates metallothionein gene transcription by altering the conformation of a specific DNA binding protein. Cell. 1988 Nov 18;55(4):705–717. doi: 10.1016/0092-8674(88)90229-2. [DOI] [PubMed] [Google Scholar]
- Hai T. W., Liu F., Coukos W. J., Green M. R. Transcription factor ATF cDNA clones: an extensive family of leucine zipper proteins able to selectively form DNA-binding heterodimers. Genes Dev. 1989 Dec;3(12B):2083–2090. doi: 10.1101/gad.3.12b.2083. [DOI] [PubMed] [Google Scholar]
- Hamer D. H. Metallothionein. Annu Rev Biochem. 1986;55:913–951. doi: 10.1146/annurev.bi.55.070186.004405. [DOI] [PubMed] [Google Scholar]
- Han K., Levine M. S., Manley J. L. Synergistic activation and repression of transcription by Drosophila homeobox proteins. Cell. 1989 Feb 24;56(4):573–583. doi: 10.1016/0092-8674(89)90580-1. [DOI] [PubMed] [Google Scholar]
- Huibregtse J. M., Engelke D. R., Thiele D. J. Copper-induced binding of cellular factors to yeast metallothionein upstream activation sequences. Proc Natl Acad Sci U S A. 1989 Jan;86(1):65–69. doi: 10.1073/pnas.86.1.65. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Imbert J., Zafarullah M., Culotta V. C., Gedamu L., Hamer D. Transcription factor MBF-I interacts with metal regulatory elements of higher eucaryotic metallothionein genes. Mol Cell Biol. 1989 Dec;9(12):5315–5323. doi: 10.1128/mcb.9.12.5315. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Johnson P. F., McKnight S. L. Eukaryotic transcriptional regulatory proteins. Annu Rev Biochem. 1989;58:799–839. doi: 10.1146/annurev.bi.58.070189.004055. [DOI] [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]
- Mueller P. R., Salser S. J., Wold B. Constitutive and metal-inducible protein:DNA interactions at the mouse metallothionein I promoter examined by in vivo and in vitro footprinting. Genes Dev. 1988 Apr;2(4):412–427. doi: 10.1101/gad.2.4.412. [DOI] [PubMed] [Google Scholar]
- Nardelli J., Gibson T. J., Vesque C., Charnay P. Base sequence discrimination by zinc-finger DNA-binding domains. Nature. 1991 Jan 10;349(6305):175–178. doi: 10.1038/349175a0. [DOI] [PubMed] [Google Scholar]
- Pugh B. F., Tjian R. Mechanism of transcriptional activation by Sp1: evidence for coactivators. Cell. 1990 Jun 29;61(7):1187–1197. doi: 10.1016/0092-8674(90)90683-6. [DOI] [PubMed] [Google Scholar]
- Roman C., Platero J. S., Shuman J., Calame K. Ig/EBP-1: a ubiquitously expressed immunoglobulin enhancer binding protein that is similar to C/EBP and heterodimerizes with C/EBP. Genes Dev. 1990 Aug;4(8):1404–1415. doi: 10.1101/gad.4.8.1404. [DOI] [PubMed] [Google Scholar]
- Safer B., Cohen R. B., Garfinkel S., Thompson J. A. DNA affinity labeling of adenovirus type 2 upstream promoter sequence-binding factors identifies two distinct proteins. Mol Cell Biol. 1988 Jan;8(1):105–113. doi: 10.1128/mcb.8.1.105. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scholer H., Haslinger A., Heguy A., Holtgreve H., Karin M. In vivo competition between a metallothionein regulatory element and the SV40 enhancer. Science. 1986 Apr 4;232(4746):76–80. doi: 10.1126/science.3006253. [DOI] [PubMed] [Google Scholar]
- Schöler H. R., Hatzopoulos A. K., Balling R., Suzuki N., Gruss P. A family of octamer-specific proteins present during mouse embryogenesis: evidence for germline-specific expression of an Oct factor. EMBO J. 1989 Sep;8(9):2543–2550. doi: 10.1002/j.1460-2075.1989.tb08392.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Searle P. F., Stuart G. W., Palmiter R. D. Building a metal-responsive promoter with synthetic regulatory elements. Mol Cell Biol. 1985 Jun;5(6):1480–1489. doi: 10.1128/mcb.5.6.1480. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Searle P. F. Zinc dependent binding of a liver nuclear factor to metal response element MRE-a of the mouse metallothionein-I gene and variant sequences. Nucleic Acids Res. 1990 Aug 25;18(16):4683–4690. doi: 10.1093/nar/18.16.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Seguin C., Hamer D. H. Regulation in vitro of metallothionein gene binding factors. Science. 1987 Mar 13;235(4794):1383–1387. doi: 10.1126/science.3103216. [DOI] [PubMed] [Google Scholar]
- Stuart G. W., Searle P. F., Chen H. Y., Brinster R. L., Palmiter R. D. A 12-base-pair DNA motif that is repeated several times in metallothionein gene promoters confers metal regulation to a heterologous gene. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7318–7322. doi: 10.1073/pnas.81.23.7318. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stuart G. W., Searle P. F., Palmiter R. D. Identification of multiple metal regulatory elements in mouse metallothionein-I promoter by assaying synthetic sequences. 1985 Oct 31-Nov 6Nature. 317(6040):828–831. doi: 10.1038/317828a0. [DOI] [PubMed] [Google Scholar]
- Szczypka M. S., Thiele D. J. A cysteine-rich nuclear protein activates yeast metallothionein gene transcription. Mol Cell Biol. 1989 Feb;9(2):421–429. doi: 10.1128/mcb.9.2.421. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Séguin C. A nuclear factor requires Zn2+ to bind a regulatory MRE element of the mouse gene encoding metallothionein-1. Gene. 1991 Jan 15;97(2):295–300. doi: 10.1016/0378-1119(91)90066-k. [DOI] [PubMed] [Google Scholar]
- Séguin C., Felber B. K., Carter A. D., Hamer D. H. Competition for cellular factors that activate metallothionein gene transcription. Nature. 1984 Dec 20;312(5996):781–785. doi: 10.1038/312781a0. [DOI] [PubMed] [Google Scholar]
- Séguin C., Prévost J. Detection of a nuclear protein that interacts with a metal regulatory element of the mouse metallothionein 1 gene. Nucleic Acids Res. 1988 Nov 25;16(22):10547–10560. doi: 10.1093/nar/16.22.10547. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thiele D. J. ACE1 regulates expression of the Saccharomyces cerevisiae metallothionein gene. Mol Cell Biol. 1988 Jul;8(7):2745–2752. doi: 10.1128/mcb.8.7.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Westin G., Schaffner W. A zinc-responsive factor interacts with a metal-regulated enhancer element (MRE) of the mouse metallothionein-I gene. EMBO J. 1988 Dec 1;7(12):3763–3770. doi: 10.1002/j.1460-2075.1988.tb03260.x. [DOI] [PMC free article] [PubMed] [Google Scholar]