Open AccessCrystallization and preliminary crystallographic studies of putative RNA 3′‐terminal phosphate cyclase from the crenarchaeon Sulfolobus tokodaii
Author(s) -
Shimizu Satoru,
Ohki Masanori,
Okubo Nami,
Suzuki Kaoru,
Tsunoda Masaru,
Sekiguchi Takeshi,
Takénaka Akio
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109014663
Subject(s) - rna , sulfolobus , rna splicing , crystal structure , biology , cyclase , crystallography , enzyme , biochemistry , chemistry , archaea , gene
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 Å 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 ( P 3 1 and P 4 2 ), with the former space group displaying two distinct packing modes.