Zinc Ions Are Differentially Required for the Transcription of Ribosomal 5S RNA and TRNA in a HeLa-cell Extract

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1984 Dec 11

PMID 6440120

Citations 2

Authors

E Wingender

D Dilloo

K H. Seifart

Affiliations

Soon will be listed here.

Abstract

Chelation of divalent cations by 5 mM EDTA and subsequent removal by dialysis from a cytoplasmic HeLa cell extract leads to a complete loss of 5S rRNA transcription without affecting tRNA synthesis. Transcription complexes for 5S RNA can no longer be assembled in such a zinc-depleted extract and this ability can be fully restored only by the re-addition of 5 microM zinc. Reconstitution experiments with isolated protein fractions show that transcription factor A from HeLa-cells requires zinc to exert its specific function. Pre-formation of transcription complexes partially protects the metal ion against removal by chelation even in the presence of 1.8 M KCl. These results indicate that the zinc ions are bound to mammalian transcription factor IIIA which, in a transcription complex, binds very strongly to the 5S RNA gene. Cation depletion with 75 mM EDTA also suppresses tRNA transcription; an effect which is reversible by zinc addition. We conclude that beside for the binding of TF IIIA, zinc is also bound with a different affinity to a transcription component common to 5S and tRNA synthesis, most likely polymerase III itself.

Citing Articles

Drawbacks of Dialysis Procedures for Removal of EDTA.

Monico A, Martinez-Senra E, Canada F, Zorrilla S, Perez-Sala D PLoS One. 2017; 12(1):e0169843.

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