Kinetic Basis of the Bifunctionality of SsoII DNA Methyltransferase

Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry
N A TimofeyevaE A Kubareva

Abstract

Type II restriction⁻modification (RM) systems are the most widespread bacterial antiviral defence mechanisms. DNA methyltransferase SsoII (M.SsoII) from a Type II RM system SsoII regulates transcription in its own RM system in addition to the methylation function. DNA with a so-called regulatory site inhibits the M.SsoII methylation activity. Using circular permutation assay, we show that M.SsoII monomer induces DNA bending of 31° at the methylation site and 46° at the regulatory site. In the M.SsoII dimer bound to the regulatory site, both protein subunits make equal contributions to the DNA bending, and both angles are in the same plane. Fluorescence of TAMRA, 2-aminopurine, and Trp was used to monitor conformational dynamics of DNA and M.SsoII under pre-steady-state conditions by stopped-flow technique. Kinetic data indicate that M.SsoII prefers the regulatory site to the methylation site at the step of initial protein⁻DNA complex formation. Nevertheless, in the presence of S-adenosyl-l-methionine, the induced fit is accelerated in the M.SsoII complex with the methylation site, ensuring efficient formation of the catalytically competent complex. The presence of S-adenosyl-l-methionine and large amount of the methylation site...Continue Reading

References

Sep 1, 1974·Proceedings of the National Academy of Sciences of the United States of America·V HershfieldD R Helinski
Dec 1, 1983·Antimicrobial Agents and Chemotherapy·G T Javor
Jul 23, 1996·Proceedings of the National Academy of Sciences of the United States of America·R A GarciaN O Reich
Feb 1, 1997·Biological & Pharmaceutical Bulletin·M MiyaharaY Yoshida
May 21, 1998·Nucleic Acids Research·I ShilovA Karyagina
Feb 15, 2000·The Journal of Biological Chemistry·W M LindstromN O Reich
Aug 10, 2000·Nucleic Acids Research·T RaskóA Kiss
Dec 5, 2000·The Journal of Biological Chemistry·G VilkaitisS Klimasauskas
Apr 3, 2001·The Journal of Biological Chemistry·G VilkaitisS Klimasauskas
Dec 1, 2001·The Journal of Biological Chemistry·Chittoor P SwaminathanAvadhesha Surolia
Jun 8, 2002·Journal of Molecular Biology·Nilesh K Banavali, Alexander D MacKerell
Jun 13, 2002·Nucleic Acids Research·Umesh T Sankpal, Desirazu N Rao
Feb 15, 2003·Proceedings of the National Academy of Sciences of the United States of America·Andrej BarbicDonald M Crothers
Jan 18, 2005·Nucleic Acids Research·Egle Merkiene, Saulius Klimasauskas
Feb 26, 2005·Biophysical Journal·Jay R UnruhGeorge S Wilson
Jun 7, 2005·Trends in Microbiology·Mya Breitbart, Forest Rohwer
Mar 1, 2007·Acta Crystallographica. Section D, Biological Crystallography·Mike CarsonLawrence J Delucas
May 2, 2007·Proceedings of the National Academy of Sciences of the United States of America·Caroline A DavisRuth M Saecker
Aug 21, 2007·Nature·Jared B ParkerJames T Stivers
Sep 15, 2007·Nature Reviews. Microbiology·Curtis A Suttle
Jan 31, 2009·FEMS Microbiology Reviews·Mario JuhasDerrick W Crook
Feb 25, 2009·Journal of Biomolecular Structure & Dynamics·N A TimofeyevaO S Fedorova

❮ Previous
Next ❯

Methods Mentioned

BETA
Fluorescence
Assay
electrophoresis
PCR
fluorescence titration
footprinting

Software Mentioned

BLAST
ProtParam
OriginPro
ImageQuant
DynaFit
OriginLab
OligoAnalyzer

Related Concepts

Related Feeds

Antivirals

Antivirals are medications that are used specifically for treating viral infections. Discover the latest research on antivirals here.

Antivirals (ASM)

Antivirals are medications that are used specifically for treating viral infections. Discover the latest research on antivirals here.