Enhancement of transcriptional activity of T7 RNA polymerase by guanidine hydrochloride.
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.
Das
M
M
Biophysics Division, Saha Institute of Nuclear Physics, Calcutta, India. milib@hp1.saha.ernet.in
Dasgupta
D
D
D017123:Bacteriophage T7 / Q000201:enzymology*; D002942:Circular Dichroism; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D019791:Guanidine / Q000494:pharmacology*; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary; D013050:Spectrometry, Fluorescence; D014158:Transcription, Genetic / Q000187:drug effects*; D014357:Trypsin / Q000378:metabolism; D014508:Urea / Q000494:pharmacology; D014764:Viral Proteins
D014764:Viral Proteins; D014508:Urea; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D014357:Trypsin; D019791:Guanidine