title: Crystallization and preliminary X-ray diffraction analysis of an archaeal tRNA-modification enzyme, TiaS, complexed with tRNA(Ile2) and ATP.
abstract: The cytidine at the first anticodon position of archaeal tRNA(Ile2), which decodes the isoleucine AUA codon, is modified to 2-agmatinylcytidine (agm(2)C) to guarantee the fidelity of protein biosynthesis. This post-transcriptional modification is catalyzed by tRNA(Ile)-agm(2)C synthetase (TiaS) using ATP and agmatine as substrates. Archaeoglobus fulgidus TiaS was overexpressed in Escherichia coli cells and purified. tRNA(Ile2) was prepared by in vitro transcription with T7 RNA polymerase. TiaS was cocrystallized with both tRNA(Ile2) and ATP by the vapour-diffusion method. The crystals of the TiaS-tRNA(Ile2)-ATP complex diffracted to 2.9 Å resolution using synchrotron radiation at the Photon Factory. The crystals belonged to the primitive hexagonal space group P3(2)21, with unit-cell parameters a = b = 131.1, c = 86.6 Å. The asymmetric unit is expected to contain one TiaS-tRNA(Ile2)-ATP complex, with a Matthews coefficient of 2.8 Å(3) Da(-1) and a solvent content of 61%.
authors.affiliation: Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba-shi, Ibaraki 305-8566, Japan.
authors.forename: Takuo
authors.initials: T
authors.lastname: Osawa
authors.forename: Hideko
authors.initials: H
authors.lastname: Inanaga
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kimura
authors.forename: Naohiro
authors.initials: N
authors.lastname: Terasaka
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Suzuki
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Numata
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry* / Q000378:metabolism; D019608:Archaeoglobales / Q000201:enzymology*; D003460:Crystallization; D018360:Crystallography, X-Ray; D007533:Isoleucine-tRNA Ligase / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D012355:RNA, Transfer, Ile / Q000737:chemistry* / Q000378:metabolism
chemical_list: D012355:RNA, Transfer, Ile; D000255:Adenosine Triphosphate; D007533:Isoleucine-tRNA Ligase
references.citation: RNA. 2008 Jan;14(1):117-26
references.pmid: 17998287
references.citation: Nat Chem Biol. 2010 Apr;6(4):277-82
references.pmid: 20139989
references.citation: EMBO J. 1998 Sep 1;17(17):5227-37
references.pmid: 9724658
references.citation: Nature. 2006 Jul 27;442(7101):419-24
references.pmid: 16871210
references.citation: Proc Natl Acad Sci U S A. 2010 Feb 16;107(7):2872-7
references.pmid: 20133752
references.citation: EMBO J. 2009 May 6;28(9):1362-73
references.pmid: 19322199
references.citation: J Mol Biol. 2006 Feb 10;356(1):97-110
references.pmid: 16343540
references.citation: Nature. 2009 Oct 22;461(7267):1144-8
references.pmid: 19847269
references.citation: Methods Enzymol. 1997;276:307-26
references.pmid: 27754618
references.citation: Nucleic Acids Res. 2003 Jan 15;31(2):532-50
references.pmid: 12527760