TY - JOUR
T1 - Expression, purification, crystallization and preliminary X-ray analysis of ORF60, the small subunit (R2) of ribonucleotide reductase from Kaposi's sarcoma-associated herpesvirus (KSHV)
AU - Gurmu, Daniel
AU - Dahlroth, Sue-Li
AU - Haas, Juergen
AU - Nordlund, Par
AU - Erlandsen, Heidi
PY - 2010/6
Y1 - 2010/6
N2 - Ribonucleotide reductase (RNR) is responsible for converting ribonucleotides to deoxyribonucleotides, which are the building blocks of DNA. The enzyme is present in all life forms as well as in some large DNA viruses such as herpesviruses. The alpha-herpesviruses and gamma-herpesviruses encode two class Ia RNR subunits, R1 and R2, while the beta-herpesvirus subfamily only encode an inactive R1 subunit. Here, the crystallization of the R2 subunit of RNR encoded by the ORF60 gene from the oncovirus Kaposi's sarcoma-associated gamma-herpesvirus (KSHV) is reported. These are the first crystals of a viral R2 subunit; the use of in situ proteolysis with chymotrypsin and the addition of hexamine cobalt(III) chloride that were necessary to obtain crystals are described. Optimization of the crystallization conditions yielded crystals that diffracted to 2.0 angstrom resolution. The crystals belonged to space group P2(1), with unit-cell parameters a = 63.9, b = 71.2, c = 71.8 angstrom, alpha = 90, beta = 106.7, gamma = 90 degrees. The data set collected was 95.3% complete, with an R-merge of 9.6%. There are two molecules in the asymmetric unit, corresponding to a solvent content of 43.4%.
AB - Ribonucleotide reductase (RNR) is responsible for converting ribonucleotides to deoxyribonucleotides, which are the building blocks of DNA. The enzyme is present in all life forms as well as in some large DNA viruses such as herpesviruses. The alpha-herpesviruses and gamma-herpesviruses encode two class Ia RNR subunits, R1 and R2, while the beta-herpesvirus subfamily only encode an inactive R1 subunit. Here, the crystallization of the R2 subunit of RNR encoded by the ORF60 gene from the oncovirus Kaposi's sarcoma-associated gamma-herpesvirus (KSHV) is reported. These are the first crystals of a viral R2 subunit; the use of in situ proteolysis with chymotrypsin and the addition of hexamine cobalt(III) chloride that were necessary to obtain crystals are described. Optimization of the crystallization conditions yielded crystals that diffracted to 2.0 angstrom resolution. The crystals belonged to space group P2(1), with unit-cell parameters a = 63.9, b = 71.2, c = 71.8 angstrom, alpha = 90, beta = 106.7, gamma = 90 degrees. The data set collected was 95.3% complete, with an R-merge of 9.6%. There are two molecules in the asymmetric unit, corresponding to a solvent content of 43.4%.
U2 - 10.1107/S1744309110016052
DO - 10.1107/S1744309110016052
M3 - Article
SN - 1744-3091
VL - 66
SP - 734
EP - 737
JO - Acta Crystallographica Section F Structural Biology and Crystallization Communications
JF - Acta Crystallographica Section F Structural Biology and Crystallization Communications
ER -