Crystallization of a member of the recFOR DNA repair pathway, RecO, with and without bound oligonucleotide.
RecFOR proteins are important for DNA repair by homologous recombination in bacteria. The RecO protein from Thermus thermophilus was cloned and purified, and its binding to oligonucleotides was characterized. The protein was crystallized alone and in complex with a 14-mer oligonucleotide. Both crystal forms grow under different crystallization conditions in the same space group, P3(1)21 or P3(2)21, with almost identical unit-cell parameters. Complete data sets were collected to 2.8 and 2.5 A for RecO alone and for the RecO-oligonucleotide complex, respectively. Visual comparison of the diffraction patterns between the two crystal forms and calculation of an R(merge) of 33.9% on F indicate that one of the crystal forms is indeed a complex of RecO with bound oligonucleotide.
Aono
Shelly
S
Physical Biosciences Division at E. O. Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA.
Hartsch
Thomas
T
Schulze-Gahmen
Ursula
U
D002852:Chromatography, Ion Exchange; D003460:Crystallization; D004260:DNA Repair / Q000235:genetics*; D004269:DNA, Bacterial / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000302:isolation & purification; D009841:Oligonucleotides / Q000737:chemistry*; D016366:Open Reading Frames / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry* / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016963:Thermus thermophilus / Q000201:enzymology / Q000235:genetics; D014961:X-Ray Diffraction
D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D009841:Oligonucleotides; C081006:RecO protein, E coli; D011994:Recombinant Proteins