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Induction of Kidney Metallothionein and Metallothionein Messenger RNA by Zinc and Cadmium^1,2,

Department of Nutrition, Rutgers University, The State University of New Jersey, New Brunswick, NJ 08903

The influence of a 7.7-µmole (0.5-mg) dose of parenteral zinc on the synthesis of metallothionein in rat kidney was examined. The amount of zinc bound to metallothionein was maximal 9–15 hours after zinc administration. Pulse labeling with [³⁵S]cystine showed the rate of renal metallothionein was stimulated to a maximum 6 hours after zinc administration and declined thereafter. Total RNA was extracted by the guanidine thiocyanate procedure from kidneys that had been flash frozen in liquid N₂. Polyadenylated RNA (mRNA) was isolated by oligo (dT)-cellulose chromatography. The mRNA was translated in a wheat germ system and newly synthesized metallothionein was isolated by activated thiol-Sepharose 4B chromatography. Metallothionein mRNA activity nearly doubled after zinc administration and was closely correlated with the enhancement in synthetic rate of this protein in kidney. Actinomycin D administered prior to either zinc or cadmium completely blocked both the stimulation of metallothionein synthesis and mRNA activity found in kidney in response to these metals. The results suggest renal metallothionein is induced by zinc or cadmium through a mechanism that requires altered expression of the metallothionein gene(s).

KEY WORDS: • kidney • metallothionein induction • mRNA • zinc • cadmium • protein synthesis

¹ Supported in part by NIH Grants AM 18555 from the National Institute of Arthritis, Metabolism and Digestive Diseases and ES 00777 from the National Institute of Environmental Health Sciences, Projects 14111 and 14112 of the New Jersey Agricultural Experiment Station and the Charles and Johanna Busch Fund of Rutgers University.

² Presented in part at the American Institute of Nutrition Meeting, Atlanta, GA, April 1981. Fed. Proc. 40, 937, abs. #4010.

³ Author to whom all inquiries and reprint requests should be addressed. Present address: Department of Food Science and Human Nutrition, University of Florida, Gainesville, FL 32611.

Manuscript received 8 October 1981.