Purification, crystallization and preliminary X-ray diffraction studies of the archaeal virus resolvase SIRV2.
The Holliday junction (or four-way junction) is the universal DNA intermediate whose interaction with resolving proteins is one of the major events in the recombinational process. These proteins, called DNA junction-resolving enzymes or resolvases, bind to the junction and catalyse DNA cleavage, promoting the release of two DNA duplexes. SIRV2 Hjc, a viral resolvase infecting a thermophylic archaeon, has been cloned, expressed and purified. Crystals have been obtained in space group C2, with unit-cell parameters a = 147.8, b = 99.9, c = 87.6, beta = 109.46 degrees, and a full data set has been collected at 3.4 A resolution. The self-rotation function indicates the presence of two dimers in the asymmetric unit and a high solvent content (77%). Molecular-replacement trials using known similar resolvase structures have so far been unsuccessful, indicating possible significant structural rearrangements.
Ennifar
Eric
E
European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117 Heidelberg, Germany.
Basquin
Jerôme
J
Birkenbihl
Rainer
R
Suck
Dietrich
D
D000595:Amino Acid Sequence; D023641:Archaeal Viruses / Q000201:enzymology* / Q000378:metabolism; D001483:Base Sequence; D003460:Crystallization; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D045565:Holliday Junction Resolvases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D023642:Rudiviridae / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D014961:X-Ray Diffraction
D045565:Holliday Junction Resolvases