title: Signal peptides bind and aggregate RNA. An alternative explanation for GTPase inhibition in the signal recognition particle.
abstract: N-terminal signal sequences can direct nascent protein chains to the inner membrane of prokaryotes and the endoplasmic reticulum of eukaryotes by interacting with the signal recognition particle. In this study, we show that isolated peptides corresponding to several bacterial signal sequences inhibit the GTPase activity of the Escherichia coli signal recognition particle, as previously reported (Miller, J. D., Bernstein, H. D., and Walter, P. (1994) Nature 367, 657-659), but not by the direct mechanism proposed. Instead, isolated signal peptides bind nonspecifically to the RNA component and aggregate the entire signal recognition particle, leading to a loss of its intrinsic GTPase activity. Surprisingly, only "functional" peptide sequences aggregate RNA; the peptides in general use as "nonfunctional" negative controls (e.g. those with deletions or charged substitutions within the hydrophobic core), are sufficiently different in physical character that they do not aggregate RNA and thus have no effect on the GTPase activity of the signal recognition particle. We propose that the reported effect of functional signal peptides on the GTPase activity of the signal recognition particle is an artifact of the high peptide concentrations and low salt conditions used in these in vitro studies and that signal sequences at the N terminus of nascent chains in vivo do not exhibit this activity.
authors.affiliation: Department of Biochemistry & Molecular Biology, University of Massachusetts, Amherst, Massachusetts 01003, USA.
authors.forename: J F
authors.initials: JF
authors.lastname: Swain
authors.forename: L M
authors.initials: LM
authors.lastname: Gierasch
mesh_terms: D020558:GTP Phosphohydrolases / Q000037:antagonists & inhibitors*; D011485:Protein Binding; D021382:Protein Sorting Signals; D012313:RNA / Q000378:metabolism*; D018271:Signal Recognition Particle; D055672:Static Electricity
chemical_list: D021382:Protein Sorting Signals; D018271:Signal Recognition Particle; D012313:RNA; D020558:GTP Phosphohydrolases