title: Purification and characterization of ribonuclease M and mRNA degradation in Escherichia coli.
abstract: A previously unreported endoribonuclease has been identified in Escherichia coli, which has a preference for hydrolysis of pyrimidine-adenosine (Pyd-Ado) bonds in RNA. It was purified about 7000-fold to give a single band after SDS/polyacrylamide gel electrophoresis; the eluted protein gave the same RNase specificity. The sizes of the native and denatured enzymes agreed suggesting that the enzyme exists as a monomer of approximately 26 kDa. It is called RNase M. The only other reported broadly specific endoribonuclease in E. coli is RNase I, a periplasmic enzyme. Based on differences in charge, heat stability and substrate specificity, it was clear that RNase M is not RNase I. The specificity of RNase M was remarkably similar to that of pancreatic RNase A even though the two enzymes differ in charge characteristics and size. Earlier studies had shown that mRNA from the lactose operon of E. coli is hydrolyzed in vivo primarily between Pyd-Ado bonds [Cannistraro et al. (1986) J. Mol. Biol. 192, 257-274] We propose that this major RNase activity accounts for these cleavages observed in vivo and that it is the endonuclease for mRNA degradation in E. coli.
authors.affiliation: Department of Microbiology and Immunology, Washington University School of Medicine, St. Louis, MO 63110.
authors.forename: V J
authors.initials: VJ
authors.lastname: Cannistraro
authors.forename: D
authors.initials: D
authors.lastname: Kennell
mesh_terms: D001483:Base Sequence; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004722:Endoribonucleases / Q000302:isolation & purification* / Q000378:metabolism; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology*; D007700:Kinetics; D012333:RNA, Messenger / Q000378:metabolism*; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D012333:RNA, Messenger; D004722:Endoribonucleases; C022538:ribonuclease M; D012259:Ribonuclease, Pancreatic