title: In Vitro/In Vivo Production of tRNA for X-Ray Studies.
abstract: tRNAs occupy a central role in the cellular life, and they are involved in a broad range of biological processes that relies on their interaction with proteins and RNA. Crystallization and structure resolution of tRNA or/and tRNA/partner complexes can yield in valuable information on structural organizations of key elements of cellular machinery. However, crystallization of RNA, is often challenging. Here we review two methods to produce and purify tRNA in quantity and quality to perform X-ray studies.
authors.affiliation: Laboratoire de Cristallographie et RMN Biologiques, CNRS, Paris Sorbonne Cité, 4 avenue de l'Observatoire, Paris, 75006, France.
authors.forename: Clément
authors.initials: C
authors.lastname: Dégut
authors.forename: Alexandre
authors.initials: A
authors.lastname: Monod
authors.forename: Franck
authors.initials: F
authors.lastname: Brachet
authors.forename: Thibaut
authors.initials: T
authors.lastname: Crépin
authors.forename: Carine
authors.initials: C
authors.lastname: Tisné
mesh_terms: D001483:Base Sequence; D002845:Chromatography; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010957:Plasmids; D012343:RNA, Transfer / Q000737:chemistry*; D015203:Reproducibility of Results; D012260:Ribonucleases / Q000737:chemistry; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000737:chemistry; D014965:X-Rays
chemical_list: D014764:Viral Proteins; D012343:RNA, Transfer; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D012260:Ribonucleases