Hfq is necessary for regulation by the untranslated RNA DsrA.
DsrA is an 85-nucleotide, untranslated RNA that has multiple regulatory activities at 30 degrees C. These activities include the translational regulation of RpoS and H-NS, global transcriptional regulators in Escherichia coli. Hfq is an E. coli protein necessary for the in vitro and in vivo replication of the RNA phage Qbeta. Hfq also plays a role in the degradation of numerous RNA transcripts. Here we show that an hfq mutant strain is defective for DsrA-mediated regulation of both rpoS and hns. The defect in rpoS expression can be partially overcome by overexpression of DsrA. Hfq does not regulate the transcription of DsrA, and DsrA does not alter the accumulation of Hfq. However, in an hfq mutant, chromosome-expressed DsrA was unstable (half-life of 1 min) and truncated at the 3' end. When expressed from a multicopy plasmid, DsrA was stable in both wild-type and hfq mutant strains, but it had only partial activity in the hfq mutant strain. Purified Hfq binds DsrA in vitro. These results suggest that Hfq acts as a protein cofactor for the regulatory activities of DsrA by either altering the structure of DsrA or forming an active RNA-protein complex.
Sledjeski
D D
DD
Department of Microbiology and Immunology, Medical College of Ohio, Toledo, Ohio 43614, USA. dsledjeski@mco.edu
Whitman
C
C
Zhang
A
A
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism*; D002876:Chromosomes, Bacterial / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D035001:Host Factor 1 Protein; D034982:Integration Host Factors; D007763:Lac Operon / Q000235:genetics; D009154:Mutation; D010957:Plasmids / Q000235:genetics; D014176:Protein Biosynthesis; D058727:RNA, Small Untranslated; D022661:RNA, Untranslated / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012808:Sigma Factor / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; C405431:DsrA RNA, E coli; D029968:Escherichia coli Proteins; C031908:H-NS protein, bacteria; D035001:Host Factor 1 Protein; D034982:Integration Host Factors; D058727:RNA, Small Untranslated; D022661:RNA, Untranslated; D011993:Recombinant Fusion Proteins; D012808:Sigma Factor; C440502:integration host factor, E coli; C069567:sigma factor KatF protein, Bacteria; C067572:RcsA protein, E coli