An E. coli ribonucleoprotein containing 4.5S RNA resembles mammalian signal recognition particle.
The signal recognition particle (SRP) plays a central role in directing the export of nascent proteins from the cytoplasm of mammalian cells. An SRP-dependent translocation machinery in bacteria has not been demonstrated in previous genetic and biochemical studies. Sequence comparisons, however, have identified (i) a gene in Escherichia coli (ffh) whose product is homologous to the 54-kilodalton subunit (SRP54) of SRP, and (ii) an RNA encoded by the ffs gene (4.5S RNA) that shares a conserved domain with the 7SL RNA of SRP. An antiserum to Ffh precipitated 4.5S RNA from E. coli extracts, implying that the two molecules reside in a complex. The 4.5S RNA can also bind to SRP54 and can replace 7SL RNA in an enzymatic assay. The product of a dominant mutation in the ffs gene (4.5S RNAdl1) is also coprecipitated by the antiserum to Ffh protein and is lethal when expressed from an inducible promoter. After induction of 4.5S RNAdl1, the earliest observed phenotype was a permanent induction of the heat shock response, suggesting that there was an accumulation of aberrant proteins in the cytoplasm. Late after induction, translocation of beta-lactamase was impaired; this may be an indirect effect of heat shock, however, because translocation of ribose binding protein or of the porin, OmpA, was unaffected. An unusual separation of the inner and outer membranes, suggestive of a defect in cell envelope, was also observed. Protein synthesis did not cease until very late, an indication that 4.5S RNA probably does not have a direct role in this process.
Department of Biochemistry and Biophysics, University of California Medical School, San Francisco 94143-0448.
M A
MA
Poritz
H D
HD
Bernstein
K
K
Strub
D
D
Zopf
H
H
Wilhelm
P
P
Walter
D001426:Bacterial Proteins / Q000378:metabolism; D001483:Base Sequence; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics*; D020558:GTP Phosphohydrolases / Q000378:metabolism; D005798:Genes, Bacterial; D006360:Heat-Shock Proteins / Q000096:biosynthesis; D006358:Hot Temperature; D007163:Immunosorbent Techniques; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D008969:Molecular Sequence Data; D009154:Mutation; D021382:Protein Sorting Signals / Q000378:metabolism; D012329:RNA, Bacterial / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012261:Ribonucleoproteins / Q000235:genetics* / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D018271:Signal Recognition Particle
D001426:Bacterial Proteins; D006360:Heat-Shock Proteins; D021382:Protein Sorting Signals; D012329:RNA, Bacterial; D011993:Recombinant Fusion Proteins; D012261:Ribonucleoproteins; D018271:Signal Recognition Particle; D007544:Isopropyl Thiogalactoside; D020558:GTP Phosphohydrolases