Open AccessPurification, crystallization and preliminary X‐ray diffraction studies of the archaeal virus resolvase SIRV2
Author(s) -
Ennifar Eric,
Basquin Jerôme,
Birkenbihl Rainer,
Suck Dietrich
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105011528
Subject(s) - holliday junction , tn3 transposon , dna , crystallography , cleavage (geology) , crystallization , base pair , chemistry , x ray crystallography , molecular replacement , stereochemistry , biology , diffraction , crystal structure , biochemistry , gene , dna repair , physics , mutant , paleontology , transposable element , organic chemistry , fracture (geology) , optics
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 C 2, with unit‐cell parameters a = 147.8, b = 99.9, c = 87.6, β = 109.46°, and a full data set has been collected at 3.4 Å 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.