title: Comparative 3-D modeling of tmRNA.
abstract: BACKGROUND
  Trans-translation releases stalled ribosomes from truncated mRNAs and tags defective proteins for proteolytic degradation using transfer-messenger RNA (tmRNA). This small stable RNA represents a hybrid of tRNA- and mRNA-like domains connected by a variable number of pseudoknots. Comparative sequence analysis of tmRNAs found in bacteria, plastids, and mitochondria provides considerable insights into their secondary structures. Progress toward understanding the molecular mechanism of template switching, which constitutes an essential step in trans-translation, is hampered by our limited knowledge about the three-dimensional folding of tmRNA.
  
  
  RESULTS
  To facilitate experimental testing of the molecular intricacies of trans-translation, which often require appropriately modified tmRNA derivatives, we developed a procedure for building three-dimensional models of tmRNA. Using comparative sequence analysis, phylogenetically-supported 2-D structures were obtained to serve as input for the program ERNA-3D. Motifs containing loops and turns were extracted from the known structures of other RNAs and used to improve the tmRNA models. Biologically feasible 3-D models for the entire tmRNA molecule could be obtained. The models were characterized by a functionally significant close proximity between the tRNA-like domain and the resume codon. Potential conformational changes which might lead to a more open structure of tmRNA upon binding to the ribosome are discussed. The method, described in detail for the tmRNAs of Escherichia coli, Bacillus anthracis, and Caulobacter crescentus, is applicable to every tmRNA.
  
  
  CONCLUSIONS
  Improved molecular models of biological significance were obtained. These models will guide in the design of experiments and provide a better understanding of trans-translation. The comparative procedure described here for tmRNA is easily adopted for the modeling the members of other RNA families.
authors.affiliation: Department of Animal Sciences, Auburn University, Auburn, AL 36849, USA. burksjm@auburn.edu
authors.forename: Jody
authors.initials: J
authors.lastname: Burks
authors.forename: Christian
authors.initials: C
authors.lastname: Zwieb
authors.forename: Florian
authors.initials: F
authors.lastname: Müller
authors.forename: Iwona
authors.initials: I
authors.lastname: Wower
authors.forename: Jacek
authors.initials: J
authors.lastname: Wower
mesh_terms: D001408:Bacillus anthracis / Q000235:genetics; D016935:Caulobacter crescentus / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D021621:Imaging, Three-Dimensional; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D012329:RNA, Bacterial / Q000737:chemistry*; D017423:Sequence Analysis, RNA / Q000379:methods
chemical_list: D012329:RNA, Bacterial; C096222:tmRNA