Functional substitution of the signal recognition particle 54-kDa subunit by its Escherichia coli homolog.
The 54-kDa subunit of the mammalian signal recognition particle (SRP54) binds to the signal sequences of nascent secretory and transmembrane proteins and facilitates their cotranslational targeting to the membrane translocation apparatus in the endoplasmic reticulum (ER). A 48-kDa Escherichia coli protein that shares extensive sequence similarity with SRP54 was identified in homology searches. Recent genetic experiments by Phillips and Silhavy [Phillips, G. J. & Silhavy, T. J. (1992) Nature (London) 359, 744-746] have shown that depletion of this protein, designated Ffh (fifty-four homolog), leads to a significant secretory defect in vivo. We demonstrate here that Ffh is structurally and functionally related to SRP54 by virtue of its ability to mimic closely its mammalian counterpart in several established biochemical assays, thereby suggesting that it plays a direct role in protein export. Ffh assembled efficiently with mammalian SRP components into a chimeric ribonucleoprotein ["SRP(Ffh)"] and bound at the site normally occupied by SRP54. Like SRP54, the Ffh moiety of the chimeric particle specifically recognized the signal sequence of preprolactin in a photocrosslinking assay. Moreover, Ffh could also act in concert with other SRP components to arrest elongation of preprolactin upon recognition of the signal sequence. In all of these assays, Ffh had approximately the same specific activity as SRP54. In contrast, SRP(Ffh) did not promote the translocation of preprolactin across the membrane of microsomal vesicles, suggesting that Ffh cannot mediate an interaction with a membrane component that is required for the translocation of nascent chains.
Department of Biochemistry and Biophysics, University of California Medical School, San Francisco 94143-0448.
H D
HD
Bernstein
D
D
Zopf
D M
DM
Freymann
P
P
Walter
D000818:Animals; D003432:Cross-Linking Reagents; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism; D007700:Kinetics; D008322:Mammals; D008861:Microsomes / Q000378:metabolism; D014176:Protein Biosynthesis; D021382:Protein Sorting Signals / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D012261:Ribonucleoproteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D018271:Signal Recognition Particle; D014466:Ultraviolet Rays
D003432:Cross-Linking Reagents; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D012261:Ribonucleoproteins; D018271:Signal Recognition Particle