title: Identification and analysis of Escherichia coli ribonuclease E dominant-negative mutants.
abstract: The Escherichia coli (E. coli) ribonuclease E protein (RNase E) is implicated in the degradation and processing of a large fraction of RNAs in the cell. To understand RNase E function in greater detail, we developed an efficient selection method for identifying nonfunctional RNase E mutants. A subset of the mutants was found to display a dominant-negative phenotype, interfering with wild-type RNase E function. Unexpectedly, each of these mutants contained a large truncation within the carboxy terminus of RNase E. In contrast, no point mutants that conferred a dominant-negative phenotype were found. We show that a representative dominant-negative mutant can form mixed multimers with RNase E and propose a model to explain how these mutants can block wild-type RNase E function in vivo.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Miami Miller School of Medicine, Miami, Florida 33136, USA.
authors.forename: Karoline J
authors.initials: KJ
authors.lastname: Briegel
authors.forename: Asmaa
authors.initials: A
authors.lastname: Baker
authors.forename: Chaitanya
authors.initials: C
authors.lastname: Jain
mesh_terms: D015153:Blotting, Western; D004722:Endoribonucleases / Q000235:genetics*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics*; D005799:Genes, Dominant; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics*; D020871:RNA Stability; D012329:RNA, Bacterial / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012641:Selection, Genetic; D017384:Sequence Deletion
chemical_list: D012329:RNA, Bacterial; D012333:RNA, Messenger; D004722:Endoribonucleases; C022537:ribonuclease E