PMID: 6160479Mar 11, 1980Paper

Preparation procedures of proteins and RNA influence the total reconstitution of 50S subunits from E. coli ribosomes

Nucleic Acids Research
V NowotnyR Amils

Abstract

A two-step procedure has been described for the total reconstitution of 50S ribosomal subunit from E. coli. RNA and proteins are mixed with stoichiometry of 1:1.2 and incubated at 44 degrees C in 4.0 mM Mg2+ followed by a second incubation at 50 degrees C in 20 mM Mg2+ (Dohme and Nierhaus, J. Mol. Biol. 107, 585 (1976)). A modified method recently reported makes use of an altered preparation technique for the RNA and proteins and requires an RNA to protein stoichiometry of 1:2.5 and 7.5 mM Mg2+ in the first incubation (Amils et al., Nucl. Acid Res. 5, 2455 (1978)). The latter requirements are not compatible with the findings obtained with the first procedure. A comparison of the various RNA and protein fractions from the different groups revealed that the Mg2+ dependence of reconstitution is a function of the RNA preparation, whereas the stoichiometry depends upon the technique used for isolation of the protein fraction. The different RNA preparations were compared in the electron microscope.

References

Dec 5, 1978·Journal of Molecular Biology·H Teraoka, K H Nierhaus
Jul 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·F Dohme, K H Nierhaus
Nov 15, 1976·Journal of Molecular Biology·F Dohme, K H Nierhaus
Nov 25, 1973·Journal of Molecular Biology·G Schreiner, K H Nierhaus
Jul 1, 1973·Proceedings of the National Academy of Sciences of the United States of America·K H Nierhaus, V Montejo
Dec 1, 1974·Proceedings of the National Academy of Sciences of the United States of America·K H Nierhaus, F Dohme

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