title: RNase III initiates rapid degradation of proU mRNA upon hypo-osmotic stress in Escherichia coli.
abstract: Hyper-osmotic stress strongly induces expression of the Escherichia coli proU operon encoding a high affinity uptake system for the osmoprotectants glycine betaine and proline betaine. Osmoregulation of proU takes place at the transcriptional level by upregulation of the promoter at high osmolarity and repression of transcription by the nucleoid-associated protein H-NS at low osmolarity. In the present study, we describe an additional level of proU osmoregulation that is independent of transcriptional regulation. We show that osmoregulation occurs at a post-transcriptional level involving RNase III. RNase III specifically processes the proU mRNA within a conserved secondary structure extending from position +203 to +293 of the transcript. Processing is efficient at low osmolarity, but inhibited at high osmolarity. Blocking of RNase III processing by mutation of the processing site eliminates post-transcriptional osmoregulation of proU. Further, the proU mRNA is relatively stable at high osmolarity with a half-life of approximately 65 sec. However, upon osmotic downshift, RNase III immediately processes the proU mRNA which reduces its half-life to less than 4 sec. The data suggest that the primary role of RNase III-mediated processing of proU mRNA is to ensure rapid shutdown of proU upon hypo-osmotic stress.
authors.affiliation: Institute for Genetics; University of Cologne; Cologne, Germany.
authors.forename: Kanstantsin
authors.initials: K
authors.lastname: Kavalchuk
authors.forename: Srinivasan
authors.initials: S
authors.lastname: Madhusudan
authors.forename: Karin
authors.initials: K
authors.lastname: Schnetz
mesh_terms: D003470:Culture Media / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D029968:Escherichia coli Proteins / Q000378:metabolism; D005798:Genes, Bacterial; D006207:Half-Life; D026901:Membrane Transport Proteins / Q000378:metabolism; D009690:Nucleic Acid Conformation; D009994:Osmolar Concentration; D033902:Periplasmic Binding Proteins / Q000378:metabolism; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D017354:Point Mutation; D012323:RNA Processing, Post-Transcriptional; D020871:RNA Stability; D012329:RNA, Bacterial / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D043244:Ribonuclease III / Q000235:genetics / Q000378:metabolism*; D013312:Stress, Physiological
chemical_list: D003470:Culture Media; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; C053437:PROX protein, E coli; D033902:Periplasmic Binding Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; D043244:Ribonuclease III