Isolation, crystallization and preliminary X-ray analysis of the transamidosome, a ribonucleoprotein involved in asparagine formation.
Thermus thermophilus deprived of asparagine synthetase synthesizes Asn on tRNA(Asn) via a tRNA-dependent pathway involving a nondiscriminating aspartyl-tRNA synthetase that charges Asp onto tRNA(Asn) prior to conversion of the Asp to Asn by GatCAB, a tRNA-dependent amidotransferase. This pathway also constitutes the route of Asn-tRNA(Asn) formation by bacteria and archaea deprived of asparaginyl-tRNA synthetase. The partners involved in tRNA-dependent Asn formation in T. thermophilus assemble into a ternary complex called the transamidosome. This particule produces Asn-tRNA(Asn) in the presence of free Asp, ATP and an amido-group donor. Crystals of the transamidosome from T. thermophilus were obtained in the presence of PEG 4000 in MES-NaOH buffer pH 6.5. They belonged to the primitive monoclinic space group P2(1), with unit-cell parameters a = 115.9, b = 214.0, c = 127.8 A, beta = 93.3 degrees . A complete data set was collected to 3 A resolution. Here, the isolation and crystallization of the transamidosome from T. thermophilus and preliminary crystallographic data are reported.
Bailly
Marc
M
UPR 9002 'Architecture et Réactivité de l'ARN', Université de Strasbourg, Institut de Biologie Moléculaire et Cellulaire du CNRS, 15 Rue René Descartes, F-67084 Strasbourg CEDEX, France.
Blaise
Mickael
M
Lorber
Bernard
B
Thirup
Soren
S
Kern
Daniel
D
D001216:Asparagine / Q000096:biosynthesis*; D001226:Aspartate-tRNA Ligase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D003460:Crystallization; D003625:Data Collection; D004926:Escherichia coli / Q000235:genetics; D008027:Light; D012346:RNA, Transfer, Amino Acyl / Q000235:genetics / Q000378:metabolism; D012348:RNA, Transfer, Asn / Q000096:biosynthesis*; D012261:Ribonucleoproteins / Q000302:isolation & purification* / Q000378:metabolism*; D012542:Scattering, Radiation; D013223:Statistics as Topic; D016963:Thermus thermophilus / Q000235:genetics / Q000378:metabolism; D046249:Transfer RNA Aminoacylation; D014961:X-Ray Diffraction
D012346:RNA, Transfer, Amino Acyl; D012348:RNA, Transfer, Asn; D012261:Ribonucleoproteins; D001216:Asparagine; D001226:Aspartate-tRNA Ligase; C029194:asparaginyl-tRNA synthetase