Purification, crystallization and preliminary X-ray analysis of the BseCI DNA methyltransferase from Bacillus stearothermophilus in complex with its cognate DNA

Evangelia G Kapetaniou; Dina Kotsifaki; Mary Providaki; Maria Rina; Vassilis Bouriotis; Michael Kokkinidis

doi:10.1107/S1744309106051530

Purification, crystallization and preliminary X-ray analysis of the BseCI DNA methyltransferase from Bacillus stearothermophilus in complex with its cognate DNA

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2007 Jan 1;63(Pt 1):12-4. doi: 10.1107/S1744309106051530. Epub 2006 Dec 16.

Authors

Evangelia G Kapetaniou¹, Dina Kotsifaki, Mary Providaki, Maria Rina, Vassilis Bouriotis, Michael Kokkinidis

Affiliation

¹ Institute of Molecular Biology and Biotechnology, PO Box 1527, GR-71110, Heraklion, Crete, Greece.

Abstract

The DNA methyltransferase M.BseCI from Bacillus stearothermophilus (EC 2.1.1.72), a 579-amino-acid enzyme, methylates the N6 atom of the 3' adenine in the sequence 5'-ATCGAT-3'. M.BseCI was crystallized in complex with its cognate DNA. The crystals were found to belong to the hexagonal space group P6, with unit-cell parameters a = b = 87.0, c = 156.1 A, beta = 120.0 degrees and one molecule in the asymmetric unit. Two complete data sets were collected at wavelengths of 1.1 and 2.0 A to 2.5 and 2.8 A resolution, respectively, using synchrotron radiation at 100 K.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Crystallization
Crystallography, X-Ray / methods*
DNA, Bacterial / chemistry
DNA, Bacterial / isolation & purification*
Geobacillus stearothermophilus / chemistry
Geobacillus stearothermophilus / isolation & purification*
Site-Specific DNA-Methyltransferase (Adenine-Specific) / chemistry*
Site-Specific DNA-Methyltransferase (Adenine-Specific) / isolation & purification*

Substances

DNA, Bacterial
DNA modification methylase Cla I
Site-Specific DNA-Methyltransferase (Adenine-Specific)