Crystallization and preliminary X-ray analysis of a RecB-family nuclease from the archaeon Pyrococcus abyssi.
Nucleases are required to process and repair DNA damage in living cells. One of the best studied nucleases is the RecB protein, which functions in Escherichia coli as a component of the RecBCD enzyme complex that amends double-strand breaks in DNA. Although archaea do not contain the RecBCD complex, a RecB-like nuclease from Pyrococcus abyssi has been cloned, expressed and purified. The protein was crystallized by the sitting-drop vapour-diffusion method using polyethylene glycol 8000 as the precipitant. The crystals belong to the orthorhombic space group C222(1), with unit-cell parameters a = 81.5, b = 159.8, c = 100.8 A. Self-rotation function and native Patterson map calculations revealed that there is a dimer in the asymmetric unit with its local twofold axis running parallel to the crystallographic twofold screw axis. The crystals diffracted to about 2 A and a complete native data set was collected to 2.65 A resolution.
Ren
Bin
B
Center for Structural Biochemistry, Karolinska Institute, NOVUM, S-141 57 Huddinge, Sweden. ren@csb.ki.se
Kuhn
Joëlle
J
Meslet-Cladiere
Laurence
L
Myllykallio
Hannu
H
Ladenstein
Rudolf
R
D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D005092:Exonucleases / Q000737:chemistry*; D011487:Protein Conformation; D043903:Pyrococcus abyssi / Q000201:enzymology*
D005092:Exonucleases