Crystallization and preliminary crystallographic studies of putative RNA 3'-terminal phosphate cyclase from the crenarchaeon Sulfolobus tokodaii.
RNA 3'-terminal phosphate cyclase (Rtc) is an enzyme involved in RNA splicing that converts the 3'-terminal hydroxyl group of truncated RNA to 2',3'-cyclic phosphate, which is required just before its ligation. This reaction may occur in the following two steps: (i) Rtc + ATP --> Rtc-AMP + PP(i) and (ii) RNA-N3'p + Rtc-AMP --> RNA-N>p + Rtc + AMP. In order to reveal the reaction mechanism, Rtc of Sulfolobus tokodaii (St-Rtc) overexpressed in Escherichia coli was purified and crystallized in the following states: St-Rtc, St-Rtc+Mn, St-Rtc+ATP, St-Rtc+AMP and St-Rtc-AMP. The crystals diffracted to 2.25-3.00 A resolution and preliminary solutions of their structures have been obtained by molecular replacement using the structure of a selenomethionine-labelled St-Rtc crystal which was solved in advance using the MAD method as a model. These crystals grew in two different space groups (P3(1) and P4(2)), with the former space group displaying two distinct packing modes.
Shimizu
Satoru
S
Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 226-8501 Yokohama, Japan.
Ohki
Masanori
M
Okubo
Nami
N
Suzuki
Kaoru
K
Tsunoda
Masaru
M
Sekiguchi
Takeshi
T
Takénaka
Akio
A
D000595:Amino Acid Sequence; D001051:Apoenzymes / Q000737:chemistry; D019843:Archaeal Proteins / Q000737:chemistry* / Q000302:isolation & purification; D002021:Buffers; D019615:Crenarchaeota / Q000201:enzymology; D003460:Crystallization; D019281:Dimerization; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008025:Ligases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010957:Plasmids; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012399:Rotation; D013223:Statistics as Topic; D016944:Sulfolobus / Q000201:enzymology*; D014169:Transformation, Bacterial; D014961:X-Ray Diffraction
D001051:Apoenzymes; D019843:Archaeal Proteins; D002021:Buffers; D008025:Ligases; C037451:RNA 3'-terminal phosphate cyclase