title: The 5΄ UTR of the type I toxin ZorO can both inhibit and enhance translation.
abstract: Many bacterial type I toxin mRNAs possess a long 5΄ untranslated region (UTR) that serves as the target site of the corresponding antitoxin sRNA. This is the case for the zorO-orzO type I system where the OrzO antitoxin base pairs to the 174-nucleotide zorO 5΄ UTR. Here, we demonstrate that the full-length 5΄ UTR of the zorO type I toxin hinders its own translation independent of the sRNA whereas a processed 5΄ UTR (zorO Δ28) promotes translation. The full-length zorO 5΄ UTR folds into an extensive secondary structure sequestering the ribosome binding site (RBS). Processing of the 5΄ UTR does not alter the RBS structure, but opens a large region (EAP region) located upstream of the RBS. Truncation of this EAP region impairs zorO translation, but this defect can be rescued upon exposing the RBS. Additionally, the region spanning +35 to +50 of the zorO mRNA is needed for optimal translation of zorO. Importantly, the positive and negative effects on translation imparted by the 5΄ UTR can be transferred onto a reporter gene, indicative that the 5΄ UTR can solely drive regulation. Moreover, we show that the OrzO sRNA can inhibit zorO translation via base pairing to the of the EAP region.
authors.affiliation: Department of Microbiology, University of Tennessee, Knoxville, TN 37996, USA.
authors.forename: Jia
authors.initials: J
authors.lastname: Wen
authors.affiliation: Department of Microbiology, University of Tennessee, Knoxville, TN 37996, USA.
authors.forename: John R
authors.initials: JR
authors.lastname: Harp
authors.affiliation: Department of Microbiology, University of Tennessee, Knoxville, TN 37996, USA.
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Fozo
mesh_terms: D020121:5' Untranslated Regions; D001427:Bacterial Toxins / Q000235:genetics* / Q000378:metabolism; D001665:Binding Sites; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D014176:Protein Biosynthesis; D012323:RNA Processing, Post-Transcriptional; D058727:RNA, Small Untranslated / Q000378:metabolism; D038621:Regulatory Sequences, Ribonucleic Acid; D012270:Ribosomes / Q000378:metabolism
chemical_list: D020121:5' Untranslated Regions; D001427:Bacterial Toxins; D029968:Escherichia coli Proteins; D058727:RNA, Small Untranslated; D038621:Regulatory Sequences, Ribonucleic Acid; C587788:ZorO protein, E coli