Two-step membrane binding by the bacterial SRP receptor enable efficient and accurate Co-translational protein targeting

The signal recognition particle (SRP) delivers ~30% of the proteome to the eukaryotic endoplasmic reticulum, or the bacterial plasma membrane. The precise mechanism by which the bacterial SRP receptor, FtsY, interacts with and is regulated at the target membrane remain unclear. Here, quantitative analysis of FtsY-lipid interactions at single-molecule resolution revealed a two-step mechanism in which FtsY initially contacts membrane via a Dynamic mode, followed by an SRP-induced conformational transition to a Stable mode that activates FtsY for downstream steps. Importantly, mutational analyses revealed extensive auto-inhibitory mechanisms that prevent free FtsY from engaging membrane in the Stable mode; an engineered FtsY pre-organized into the Stable mode led to indiscriminate targeting in vitro and disrupted FtsY function in vivo. Our results show that the two-step lipid-binding mechanism uncouples the membrane association of FtsY from its conformational activation, thus optimizing the balance between the efficiency and fidelity of co-translational protein targeting. DOI: http://dx.doi.org/10.7554/eLife.25885.001

• Publication year

  2017

• Venue

  eLife

• Publication date

  2017-07-28

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.7554/eLife.25885PMID 28753124PMCID 5533587
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• The bacterial SRP receptor, FtsY, is activated on binding to the translocon

  2016influential reference
• Ribosome binding induces repositioning of the signal recognition particle receptor on the translocon

  2015influential reference
• Fidelity of cotranslational protein targeting by the signal recognition particle.

  2014cited by this paper
• Signal Recognition Particle-ribosome Binding Is Sensitive to Nascent Chain Length*

  2014cited by this paper
• SecYEG activates GTPases to drive the completion of cotranslational protein targeting

  2013influential reference
• Signal recognition particle: an essential protein-targeting machine.

  2013influential reference
• Structural basis of signal sequence surveillance and selection by the SRP–FtsY complex

  2013influential reference
• Heat Shock Transcription Factor σ32 Co-opts the Signal Recognition Particle to Regulate Protein Homeostasis in E. coli

  2013cited by this paper
• Fingerloop activates cargo delivery and unloading during cotranslational protein targeting

  2013influential reference
• Advances in analysis of low signal-to-noise images link dynamin and AP2 to the functions of an endocytic checkpoint.

  2013cited by this paper
• Activated GTPase movement on an RNA scaffold drives cotranslational protein targeting

  2012influential reference
• Speed, dissipation, and error in kinetic proofreading

  2012cited by this paper
• Translation Elongation Regulates Substrate Selection by the Signal Recognition Particle*

  2012cited by this paper
• Signal sequence–independent SRP-SR complex formation at the membrane suggests an alternative targeting pathway within the SRP cycle

  2011cited by this paper
• The Crystal Structure of the Signal Recognition Particle in Complex with Its Receptor

  2011cited by this paper
• Genetic code translation displays a linear trade-off between efficiency and accuracy of tRNA selection

  2011cited by this paper
• Lipids Trigger a Conformational Switch That Regulates Signal Recognition Particle (SRP)-mediated Protein Targeting*

  2011influential reference
• Optimization of speed and accuracy of decoding in translation

  2010cited by this paper
• Lipid activation of the signal recognition particle receptor provides spatial coordination of protein targeting

  2010influential reference
• Supported Membrane Formation, Characterization, Functionalization, and Patterning for Application in Biological Science and Technology

  2010cited by this paper
• Genetic Evidence for Functional Interaction of the Escherichia coli Signal Recognition Particle Receptor with Acidic Lipids in Vivo*

  2010cited by this paper
• The action of cardiolipin on the bacterial translocon

  2010influential reference
• Sequential Checkpoints Govern Substrate Selection During Cotranslational Protein Targeting

  2010influential reference
• Two cooperating helices constitute the lipid-binding domain of the bacterial SRP receptor.

  2009cited by this paper
• Delivering proteins for export from the cytosol

  2009cited by this paper
• Multiple conformational switches in a GTPase complex control co-translational protein targeting

  2009influential reference
• Predominant membrane localization is an essential feature of the bacterial signal recognition particle receptor

  2009cited by this paper
• Enzymatic assembly of DNA molecules up to several hundred kilobases

  2009cited by this paper
• Signal Sequences Activate the Catalytic Switch of SRP RNA

  2009cited by this paper
• Rate and accuracy of bacterial protein synthesis revisited.

  2008cited by this paper
• Demonstration of a multistep mechanism for assembly of the SRP x SRP receptor complex: implications for the catalytic role of SRP RNA.

  2008cited by this paper
• SRP RNA controls a conformational switch regulating the SRP–SRP receptor interaction

  2008influential reference
• A cleavable N-terminal membrane anchor is involved in membrane binding of the Escherichia coli SRP receptor.

  2008influential reference
• Functional substitution of the transient membrane-anchor domain in Escherichia coli FtsY with an N-terminal hydrophobic segment of Streptomyces lividans FtsY.

  2008cited by this paper
• A practical guide to single-molecule FRET

  2008influential reference
• Effect of surface treatment on diffusion and domain formation in supported lipid bilayers.

  2007cited by this paper
• Escherichia coli Signal Recognition Particle Receptor FtsY Contains an Essential and Autonomous Membrane-binding Amphipathic Helix*

  2007influential reference
• Conformational changes in the GTPase modules of the signal reception particle and its receptor drive initiation of protein translocation

  2007cited by this paper
• Membrane Targeting of Ribosomes and Their Release Require Distinct and Separable Functions of FtsY*

  2007influential reference
• Structure of the GMPPNP-stabilized NG domain complex of the SRP GTPases Ffh and FtsY.

  2006cited by this paper
• Membrane binding of the bacterial signal recognition particle receptor involves two distinct binding sites

  2006influential reference
• FtsY, the bacterial signal‐recognition particle receptor, interacts functionally and physically with the SecYEG translocon

  2005influential reference
• Mechanism of Association and Reciprocal Activation of Two GTPases

  2004cited by this paper
• The Core Escherichia coli Signal Recognition Particle Receptor Contains Only the N and G Domains of FtsY

  2004cited by this paper
• Substrate twinning activates the signal recognition particle and its receptor

  2004cited by this paper
• Heterodimeric GTPase Core of the SRP Targeting Complex

  2004cited by this paper
• Crystallization of the GMPPCP complex of the NG domains of Thermus aquaticus Ffh and FtsY.

  2003cited by this paper
• Conformational change of the N-domain on formation of the complex between the GTPase domains of Thermus aquaticus Ffh and FtsY.

  2002cited by this paper
• Role of SRP RNA in the GTPase cycles of Ffh and FtsY.

  2001cited by this paper
• Putative integral membrane SRP receptors.

  2001cited by this paper
• Anionic phospholipids are involved in membrane association of FtsY and stimulate its GTPase activity

  2000influential reference
• Formation and Spreading of Lipid Bilayers on Planar Glass Supports

  1999cited by this paper
• The NG domain of the prokaryotic signal recognition particle receptor, FtsY, is fully functional when fused to an unrelated integral membrane polypeptide.

  1997cited by this paper
• Co‐translational protein targeting catalyzed by the Escherichia coli signal recognition particle and its receptor

  1997influential reference
• The beta subunit of the signal recognition particle receptor is a transmembrane GTPase that anchors the alpha subunit, a peripheral membrane GTPase, to the endoplasmic reticulum membrane

  1995cited by this paper
• Signal sequence recognition and protein targeting to the endoplasmic reticulum membrane.

  1994cited by this paper
• The accuracy of protein biosynthesis is limited by its speed: high fidelity selection by ribosomes of aminoacyl-tRNA ternary complexes containing GTP[gamma S].

  1982cited by this paper
• Kinetic proofreading: a new mechanism for reducing errors in biosynthetic processes requiring high specificity.

  1974cited by this paper
• Signal Sequences Activate the Catalytic Switch of SRP RNA

  year unknowninfluential reference

• Pig jejunal single-cell RNA landscapes revealing breed-specific immunology differentiation at various domestication stages

  2025cites this paper
• Mechanistic insights into protein biogenesis and maturation on the ribosome.

  2025cites this paper
• Unravelling the role of the a domain and N-terminal alpha-helices of FtsY in archaeal signal recognition particle.

  2025cites this paper
• The viable but non-culturable (VBNC) status of Shewanella putrefaciens (S. putrefaciens) with thermosonication (TS) treatment

  2024cites this paper
• A Trivalent Live Vaccine Elicits Cross-Species Protection Against Acute Otitis Media in a Murine Model

  2024cites this paper
• The metabolic basis of inherited neutropenias

  2023cites this paper
• Construction and development of a novel dual-gene coexpression system to promote heterologous protein secretion for Streptomyces.

  2022cites this paper
• A First Insight Into the Heat-Induced Changes in Proteomic Profiles of the Coral Symbiotic Bacterium Endozoicomonas montiporae

  2022cites this paper
• N-terminal helices and A domain of archaeal FtsY facilitate SRP54 binding and the association with cell membrane

  2022cites this paper
• Fidelity of Cotranslational Protein Targeting to the Endoplasmic Reticulum

  2021cites this paper
• Response of Endozoicomonas montiporae to heat stress and coral host lysates

  2021cites this paper
• Molecular mechanism of cargo recognition and handover by the mammalian signal recognition particle

  2021cites this paper
• Direct observation of alpha-lactalbumin, adsorption and incorporation into lipid membrane and formation of lipid/protein hybrid structures.

  2019cites this paper
• Specific structural elements of the T-box riboswitch drive the two-step binding of the tRNA ligand

  2018cites this paper