Abstract
The expression of metallothionein (MT)-1 and -2 mRNAs in rat liver following administration of Cd or Cu was investigated using specific oligonucleotides. The specificity was confirmed using a competitive prehybridization assay. Cd injection caused a biphasic induction of both isoform mRNAs, whereas Cu induced a sustained, monophasic response. Analysis of polyribosomal RNA showed that, after both Cd and Cu treatments, the recruitment of MT-1 mRNA into polyribosomes paralleled the increase in transcription, but the increase of polyribosomal MT-2 mRNA was less than that of total MT-2 mRNA. This indicates that not all the MT-2 mRNA induced was translated, suggesting that there is translational control of MT-2 mRNA expression, but not of MT-1 mRNA. This hypothesis was supported by the observation that, after Cu treatment, the induction of MT-1 protein was induced to the same extent as MT-1 mRNA, whereas the total MT protein (MT-1 + MT-2) was increased far less (7-fold) than MT-2 mRNA (30-fold).
Full Text
The Full Text of this article is available as a PDF (422.5 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Andersen R. D., Piletz J. E., Birren B. W., Herschman H. R. Levels of metallothionein messenger RNA in foetal, neonatal and maternal rat liver. Eur J Biochem. 1983 Apr 5;131(3):497–500. doi: 10.1111/j.1432-1033.1983.tb07289.x. [DOI] [PubMed] [Google Scholar]
- Andersen R. D., Taplitz S. J., Birren B. W., Bristol G., Herschman H. R. Rat metallothionein multigene family. Experientia Suppl. 1987;52:373–384. doi: 10.1007/978-3-0348-6784-9_35. [DOI] [PubMed] [Google Scholar]
- Andrews G. K. Regulation of metallothionein gene expression. Prog Food Nutr Sci. 1990;14(2-3):193–258. [PubMed] [Google Scholar]
- Blobel G., Sabatini D. Dissociation of mammalian polyribosomes into subunits by puromycin. Proc Natl Acad Sci U S A. 1971 Feb;68(2):390–394. doi: 10.1073/pnas.68.2.390. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bremner I., Beattie J. H. Metallothionein and the trace minerals. Annu Rev Nutr. 1990;10:63–83. doi: 10.1146/annurev.nu.10.070190.000431. [DOI] [PubMed] [Google Scholar]
- Bremner I., Hoekstra G., Davies N. T., Young B. W. Effect of zinc status of rats on the synthesis and degradation of copper-induced metallothioneins. Biochem J. 1978 Sep 15;174(3):883–892. doi: 10.1042/bj1740883. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bremner I. Interactions between metallothionein and trace elements. Prog Food Nutr Sci. 1987;11(1):1–37. [PubMed] [Google Scholar]
- Bremner I. Involvement of metallothionein in the hepatic metabolism of copper. J Nutr. 1987 Jan;117(1):19–29. doi: 10.1093/jn/117.1.19. [DOI] [PubMed] [Google Scholar]
- Cherian M. G., Yu S., Redman C. M. Site of synthesis of metallothionein in rat liver. Can J Biochem. 1981 Apr;59(4):301–306. doi: 10.1139/o81-041. [DOI] [PubMed] [Google Scholar]
- 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]
- Chubatsu L. S., Meneghini R. Metallothionein protects DNA from oxidative damage. Biochem J. 1993 Apr 1;291(Pt 1):193–198. doi: 10.1042/bj2910193. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fleet J. C., Golemboski K. A., Dietert R. R., Andrews G. K., McCormick C. C. Induction of hepatic metallothionein by intraperitoneal metal injection: an associated inflammatory response. Am J Physiol. 1990 Jun;258(6 Pt 1):G926–G933. doi: 10.1152/ajpgi.1990.258.6.G926. [DOI] [PubMed] [Google Scholar]
- Gray N. K., Hentze M. W. Regulation of protein synthesis by mRNA structure. Mol Biol Rep. 1994 May;19(3):195–200. doi: 10.1007/BF00986961. [DOI] [PubMed] [Google Scholar]
- Gunneberg A., Scobie G., Hayes K., Kalsheker N. Competitive assay to improve the specificity of detection of single-point mutations in alpha 1-antitrypsin deficiency. Clin Chem. 1993 Oct;39(10):2157–2162. [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]
- Hovland R., Campbell G., Pryme I., Hesketh J. The mRNAs for cyclin A, c-myc and ribosomal proteins L4 and S6 are associated with cytoskeletal-bound polysomes in HepG2 cells. Biochem J. 1995 Aug 15;310(Pt 1):193–196. doi: 10.1042/bj3100193. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Iijima Y., Fukushima T., Bhuiyan L. A., Yamada T., Kosaka F., Sato J. D. Synergistic and additive induction of metallothionein in Chang liver cells. A possible mechanism of marked induction of metallothionein by stress. FEBS Lett. 1990 Aug 20;269(1):218–220. doi: 10.1016/0014-5793(90)81158-k. [DOI] [PubMed] [Google Scholar]
- Kozak M. Structural features in eukaryotic mRNAs that modulate the initiation of translation. J Biol Chem. 1991 Oct 25;266(30):19867–19870. [PubMed] [Google Scholar]
- Lehman-McKeeman L. D., Andrews G. K., Klaassen C. D. Induction of hepatic metallothioneins determined at isoprotein and messenger RNA levels in glucocorticoid-treated rats. Biochem J. 1988 Jan 15;249(2):429–433. doi: 10.1042/bj2490429. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lehman-McKeeman L. D., Andrews G. K., Klaassen C. D. Mechanisms of regulation of rat hepatic metallothionein-I and metallothionein-II levels following administration of zinc. Toxicol Appl Pharmacol. 1988 Jan;92(1):1–9. doi: 10.1016/0041-008x(88)90221-9. [DOI] [PubMed] [Google Scholar]
- Lehman-McKeeman L. D., Andrews G. K., Klaassen C. D. Ontogeny and induction of hepatic isometallothioneins in immature rats. Toxicol Appl Pharmacol. 1988 Jan;92(1):10–17. doi: 10.1016/0041-008x(88)90222-0. [DOI] [PubMed] [Google Scholar]
- McCarthy J. E., Kollmus H. Cytoplasmic mRNA-protein interactions in eukaryotic gene expression. Trends Biochem Sci. 1995 May;20(5):191–197. doi: 10.1016/s0968-0004(00)89006-4. [DOI] [PubMed] [Google Scholar]
- McCormick C. C., Salati L. M., Goodridge A. G. Abundance of hepatic metallothionein mRNA is increased by protein-synthesis inhibitors. Evidence for transcriptional activation and post-transcriptional regulation. Biochem J. 1991 Jan 1;273(Pt 1):185–188. doi: 10.1042/bj2730185. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mehra R. K., Bremner I. Development of a radioimmunoassay for rat liver metallothionein-I and its application to the analysis of rat plasma and kidneys. Biochem J. 1983 Aug 1;213(2):459–465. doi: 10.1042/bj2130459. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Peterson M. G., Mercer J. F. Differential expression of four linked sheep metallothionein genes. Eur J Biochem. 1988 Jun 1;174(2):425–429. doi: 10.1111/j.1432-1033.1988.tb14115.x. [DOI] [PubMed] [Google Scholar]
- Piletz J. E., Andersen R. D., Birren B. W., Herschman H. R. Metallothionein synthesis in foetal, neonatal and maternal rat liver. Eur J Biochem. 1983 Apr 5;131(3):489–495. doi: 10.1111/j.1432-1033.1983.tb07288.x. [DOI] [PubMed] [Google Scholar]
- Sadhu C., Gedamu L. Metal-specific posttranscriptional control of human metallothionein genes. Mol Cell Biol. 1989 Dec;9(12):5738–5741. doi: 10.1128/mcb.9.12.5738. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sato M., Bremner I. Oxygen free radicals and metallothionein. Free Radic Biol Med. 1993 Mar;14(3):325–337. doi: 10.1016/0891-5849(93)90029-t. [DOI] [PubMed] [Google Scholar]
- Scheuhammer A. M., Cherian M. G. Quantification of metallothionein by silver saturation. Methods Enzymol. 1991;205:78–83. doi: 10.1016/0076-6879(91)05088-d. [DOI] [PubMed] [Google Scholar]
- Shapiro S. G., Squibb K. S., Markowitz L. A., Cousins R. J. Cell-free synthesis of metallothionein directed by rat liver polyadenylated messenger ribonucleic acid. Biochem J. 1978 Dec 1;175(3):833–840. doi: 10.1042/bj1750833. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wake S. A., Mercer J. F. Induction of metallothionein mRNA in rat liver and kidney after copper chloride injection. Biochem J. 1985 Jun 1;228(2):425–432. doi: 10.1042/bj2280425. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Webb M. Toxicological significance of metallothionein. Experientia Suppl. 1987;52:109–134. doi: 10.1007/978-3-0348-6784-9_6. [DOI] [PubMed] [Google Scholar]
- White K., Munro H. N. Induction of ferritin subunit synthesis by iron is regulated at both the transcriptional and translational levels. J Biol Chem. 1988 Jun 25;263(18):8938–8942. [PubMed] [Google Scholar]