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. 1974 Sep;71(9):3395–3399. doi: 10.1073/pnas.71.9.3395

RNA Polymerase Assembly In Vitro. Temperature Dependence of Reactivation of Denatured Core Enzyme

John D Harding 1,2,*, Sherman Beychok 1,2,†
PMCID: PMC433779  PMID: 4610575

Abstract

The Escherichia coli RNA polymerase core molecule, after denaturation in 6 M guanidine hydrochloride, can be completely reactivated in the absence of sigma subunit. Reactivation is temperature dependent. At 4° a renatured-inactive preparation is formed that has most of the secondary structure of the original native molecule but has a reduced sedimentation coefficient and a smaller Stokes radius and is, therefore, of lower molecular weight. Upon warming to 37° the renatured-inactive preparation is converted in a time-dependent process to the renatured-active preparation, which has the same amount of secondary structure and same molecular weight as native RNA polymerase. Since the renatured-inactive material is probably composed of subunit assemblies and can be readily reactivated, it should be useful for studying the subunit interactions and control of assembly of RNA polymerase.

Keywords: transcription, subunit enzymes, Escherichia coli

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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