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Enhancement of transcriptional activity of T7 RNA polymerase by guanidine hydrochloride
Author(s) -
Das Mili,
Dasgupta Dipak
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00458-x
Subject(s) - guanidine , proteolysis , hydrochloride , chemistry , enzyme , transcription (linguistics) , enzyme assay , biochemistry , urea , rna polymerase , polymerase , rna , rna polymerase ii , t7 rna polymerase , microbiology and biotechnology , biology , gene expression , promoter , gene , linguistics , philosophy , escherichia coli , bacteriophage
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.