Enhancement of transcriptional activity of T7 RNA polymerase by guanidine hydrochloride
Open Access
- 15 May 1998
- journal article
- Published by Wiley in FEBS Letters
- Vol. 427 (3) , 337-340
- https://doi.org/10.1016/s0014-5793(98)00458-x
Abstract
T7 RNA polymerase shows an increase in processive transcription in the presence of low concentrations of guanidine hydrochloride (GdnCl) upto 60 mM, which is not observed when the enzyme is treated with urea. Higher concentrations of the denaturant lead to a progressive loss in the processive transcriptional activity of the enzyme. We have attempted to explain the above phenomenon in terms of the structural change in the enzyme. Fluorescence and CD studies suggest that the tertiary structure of the native enzyme undergoes an alteration upon addition of low concentration of guanidine hydrochloride. This is also indicated from the decreased susceptibility of the enzyme to limited proteolysis by trypsin.Keywords
This publication has 17 references indexed in Scilit:
- Probing the conformational state of apomyoglobin by limited proteolysis 1 1Edited by P. E. WrightJournal of Molecular Biology, 1997
- Subunit Dissociation and Unfolding of Macrophage NO Synthase: Relationship between Enzyme Structure, Prosthetic Group Binding, and Catalytic FunctionBiochemistry, 1995
- A Partially Folded Intermediate during Tubulin Unfolding: Its Detection and Spectroscopic CharacterizationBiochemistry, 1995
- The Thumbs's KnuckleJournal of Molecular Biology, 1994
- Bacteriophage T7 RNA Polymerase and Its Active-site Mutants: Kinetic, Spectroscopic and Calorimetric CharacterizationJournal of Molecular Biology, 1994
- A Kinetic Method to Evaluate the Two-State Character of Solvent-Induced Protein DenaturationBiochemistry, 1994
- Crystal structure of bacteriophage T7 RNA polymerase at 3.3 Å resolutionNature, 1993
- Processivity of proteolytically modified forms of T7 RNA polymeraseBiochemistry, 1988
- Nucleotide sequence of the cloned gene for bacteriophage T7 RNA polymeraseJournal of Molecular Biology, 1981
- Recognition and initiation site for four late promoters of phage T7 is a 22-base pair DNA sequenceNature, 1979