Anionic phospholipids are involved in membrane association of FtsY and stimulate its GTPase activity

Leeuw, Erik de; Kaat, Kai te; Moser, C.; Menestrina, Gianfranco; Demel, R.A; Kruijff, Ben de; Oudega, Bauke; Luirink, Joen; Sinning, Irmgard

doi:10.1093/emboj/19.4.531

Cited by 139 publications

(171 citation statements)

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“…15 and this study), we show that the MTS is highly dynamic in solution. Previous studies showed that FtsY has a preference for anionic phospholipids (9), whereas other studies also suggested a role of zwitterionic lipids (36). The current study clarifies that only the interaction with anionic phospholipids triggers a random-coil helix transition of the MTS.…”

Section: Discussionsupporting

confidence: 70%

“…1A). The A domain acts as negative regulator of the FtsY GTPase in a lipid-free environment (9) and was suggested to participate in membrane interaction of FtsY by its N-terminal region (10). However, the A domain is not essential in Escherichia coli as a truncation variant (termed NGϩ1) is functional in vivo (11).…”

mentioning

confidence: 99%

“…However, the A domain is not essential in Escherichia coli as a truncation variant (termed NGϩ1) is functional in vivo (11). It was shown that the FtsY GTPase is activated by anionic phospholipids (9) and that the membrane interaction of FtsY is crucial for the release of RNCs from the SRP-FtsY complex (12,13), which was confirmed recently (14). Membrane interaction of E. coli FtsY depends on a conserved motif at the N terminus of the N domain, referred to as membrane-targeting sequence (MTS) (15).…”

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confidence: 99%

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Lipids Trigger a Conformational Switch That Regulates Signal Recognition Particle (SRP)-mediated Protein Targeting

Stjepanović

Kapp

Bange

et al. 2011

Journal of Biological Chemistry

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Co-translational protein targeting to the membrane is mediated by the signal recognition particle and its receptor (FtsY). Their homologous GTPase domains interact at the membrane and form a heterodimer in which both GTPases are activated. The prerequisite for protein targeting is the interaction of FtsY with phospholipids. However, the mechanism of FtsY regulation by phospholipids remained unclear. Here we show that the N terminus of FtsY (A domain) is natively unfolded in solution and define the complete membrane-targeting sequence. We show that the membrane-targeting sequence is highly dynamic in solution, independent of nucleotides and directly responds to the density of anionic phospholipids by a random coil-helix transition. This conformational switch is essential for tethering FtsY to membranes and activates the GTPase for its subsequent interaction with the signal recognition particle. Our results underline the dynamics of lipid-protein interactions and their importance in the regulation of protein targeting and translocation across biological membranes.The biogenesis of most membrane proteins and many secretory proteins depends on the signal recognition particle (SRP) 4 and the SRP receptor (SR). SRP binds to N-terminal signal sequences of nascent polypeptide chains at the ribosome (ribosome-nascent chain complexes (RNCs)) and acts as an adaptor between the ribosome and the membrane-embedded translocation channel (1-3). Interaction of SRP with SR (FtsY in bacteria and archaea and SR␣ in eukarya) specifies the target membrane and allows for the precise coordination of RNC release from SRP and its transfer to the translocation channel. Protein targeting critically depends on the two homologous GTPases present in SRP and SR forming a heterodimer (4). GTP binding to SRP and SR is required for heterodimer formation and GTP hydrolysis triggers the dissociation of the SRP-SR complex which resets the SRP system for a new round of translocation (5). Although FtsY does not contain a hydrophobic, transmembrane sequence, it was shown to be almost exclusively localized at the membrane (6). FtsY contains three domains: an N-terminal negatively charged A domain of unknown structure and function and the highly conserved N and G domains that form a structural and functional unit, the NG domain (7, 8) (see Fig. 1A). The A domain acts as negative regulator of the FtsY GTPase in a lipid-free environment (9) and was suggested to participate in membrane interaction of FtsY by its N-terminal region (10). However, the A domain is not essential in Escherichia coli as a truncation variant (termed NGϩ1) is functional in vivo (11). It was shown that the FtsY GTPase is activated by anionic phospholipids (9) and that the membrane interaction of FtsY is crucial for the release of RNCs from the SRP-FtsY complex (12, 13), which was confirmed recently (14). Membrane interaction of E. coli FtsY depends on a conserved motif at the N terminus of the N domain, referred to as membrane-targeting sequence (MTS) (15). Recently, strong genetic ...

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Section: Discussionsupporting

confidence: 70%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Lipids Trigger a Conformational Switch That Regulates Signal Recognition Particle (SRP)-mediated Protein Targeting

Stjepanović

Kapp

Bange

et al. 2011

Journal of Biological Chemistry

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“…Because full-length FlhF is a monomer with bound GDP the B domain and the nucleotide load might be involved in the regulation of dimerization. For E. coli FtsY a negative regulatory effect of the A domain on GTP hydrolysis was described (47). How and in which functional context GTP hydrolysis is activated in FlhF is not known at present.…”

Section: Resultsmentioning

confidence: 99%

The crystal structure of the third signal-recognition particle GTPase FlhF reveals a homodimer with bound GTP

Bange

Petzold

Wild

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

101

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Flagella are well characterized as the organelles of locomotion and allow bacteria to react to environmental changes. The assembly of flagella is a multistep process and relies on a complex type III export machinery located in the cytoplasmic membrane. The FlhF protein is essential for the placement and assembly of polar flagella and has been classified as a signal-recognition particle (SRP)-type GTPase. SRP GTPases appeared early in evolution and form a unique subfamily within the guanine nucleotide binding proteins with only three members: the signal sequence-binding protein SRP54, the SRP receptor FtsY, and FlhF. We report the crystal structures of FlhF from Bacillus subtilis in complex with GTP and GMPPNP. FlhF shares SRP GTPase-specific features such as the presence of an N-terminal ␣-helical domain and the I-box insertion. It forms a symmetric homodimer sequestering a composite active site that contains two head-to-tail arranged nucleotides similar to the heterodimeric SRP-targeting complex. However, significant differences to the GTPases of SRP and the SRP receptor include the formation of a stable homodimer with GTP as well as severe modifications and even the absence of motifs involved in regulation of the other two SRP GTPases. Our results provide insights into SRP GTPases and their roles in two fundamentally different protein-targeting routes that both rely on efficient protein delivery to a secretion channel.flagellum ͉ protein translocation ͉ x-ray structure analysis ͉ protein targeting

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“…This may indicate that additional factors are required for high (catalytic) rates of GTP hydrolysis. In bacteria, lipid interactions stimulate the GTPase activity of FtsY (39), and interaction of available translocase components with targeting machinery initiates GTP hydrolysis in targeting to the ER (40). In that context, the low rate of GTP hydrolysis by cpSRP54/cpFtsY in solution is likely of biological significance because high rates of GTP hydrolysis in solution would lead to a potentially deleterious futile cycle of GTP hydrolysis in stroma.…”

Section: Fig 6 Functional Analysis Of the C-terminal 26 Amino Acidsmentioning

confidence: 99%

Regulation of the GTPase Cycle in Post-translational Signal Recognition Particle-based Protein Targeting Involves cpSRP43

Goforth¹,

Peterson²,

Yuan³

et al. 2004

Journal of Biological Chemistry

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Anionic phospholipids are involved in membrane association of FtsY and stimulate its GTPase activity

Abstract: Ffh for fifty four homologue) that show homology to the eukaryotic 7S RNA and SRP54, respectively. Recent

Cited by 139 publications

References 49 publications

Lipids Trigger a Conformational Switch That Regulates Signal Recognition Particle (SRP)-mediated Protein Targeting

Lipids Trigger a Conformational Switch That Regulates Signal Recognition Particle (SRP)-mediated Protein Targeting

The crystal structure of the third signal-recognition particle GTPase FlhF reveals a homodimer with bound GTP

Regulation of the GTPase Cycle in Post-translational Signal Recognition Particle-based Protein Targeting Involves cpSRP43

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