Twin-Arginine Protein Translocation

Goosens, Vivianne J.; Dijl, Jan Maarten van

doi:10.1007/82_2016_7

Cited by 17 publications

(22 citation statements)

References 171 publications

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“…However, unlike sortase substrates, which are anchored to the peptidoglycan cell wall, ArtA substrates are covalently attached to a lipid embedded in the cytoplasmic membrane following C‐terminal processing (Abdul Halim et al ., ). Moreover, in contrast to sortase A substrates, which appear to be secreted exclusively via the general secretory (Sec) pathway, we recently determined that some ArtA substrates are translocated across the cytoplasmic membrane via the Twin Arginine Transport (Tat) pathway while others are translocated via the Sec pathway (Schneewind and Missiakas, ; Goosens and van Dijl, ; Abdul Halim et al ., ).…”

Section: Introductionmentioning

confidence: 99%

Conserved residues are critical for Haloferax volcanii archaeosortase catalytic activity: Implications for convergent evolution of the catalytic mechanisms of non‐homologous sortases from archaea and bacteria

Halim

Rodríguez

Stoltzfus

et al. 2018

Molecular Microbiology

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Proper protein anchoring is key to the biogenesis of prokaryotic cell surfaces, dynamic, resilient structures that play crucial roles in various cell processes. A novel surface protein anchoring mechanism in Haloferax volcanii depends upon the peptidase archaeosortase A (ArtA) processing C-termini of substrates containing C-terminal tripartite structures and anchoring mature substrates to the cell membrane via intercalation of lipid-modified C-terminal amino acid residues. While this membrane protein lacks clear homology to soluble sortase transpeptidases of Gram-positive bacteria, which also process C-termini of substrates whose C-terminal tripartite structures resemble those of ArtA substrates, archaeosortases do contain conserved cysteine, arginine and arginine/histidine/asparagine residues, reminiscent of His-Cys-Arg residues of sortase catalytic sites. The study presented here shows that ArtA -GFP expressed in trans complements ΔartA growth and motility phenotypes, while alanine substitution mutants, Cys (C173A), Arg (R214A) or Arg (R253A), and the serine substitution mutant for Cys (C173S), fail to complement these phenotypes. Consistent with sortase active site replacement mutants, ArtA -GFP, ArtA -GFP and ArtA -GFP cannot process substrates, while replacement of the third residue, ArtA -GFP retains some processing activity. These findings support the view that similarities between certain aspects of the structures and functions of the sortases and archaeosortases are the result of convergent evolution.

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Section: Introductionmentioning

confidence: 99%

Conserved residues are critical for Haloferax volcanii archaeosortase catalytic activity: Implications for convergent evolution of the catalytic mechanisms of non‐homologous sortases from archaea and bacteria

Halim

Rodríguez

Stoltzfus

et al. 2018

Molecular Microbiology

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“…ported across the cytoplasmic membrane via either the secretion (Sec) or the twinarginine transport (Tat) pathway (1)(2)(3). The Sec pathway, which is found in all species of eukaryotes as well as those of both prokaryotic domains, transports proteins across the cytoplasmic membrane in an unfolded conformation.…”

mentioning

confidence: 99%

ArtA-Dependent Processing of a Tat Substrate Containing a Conserved Tripartite Structure That Is Not Localized at the C Terminus

et al. 2017

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Most prokaryote-secreted proteins are transported to the cell surface using either the general secretion (Sec) or twin-arginine translocation (Tat) pathway. A majority of secreted proteins are anchored to the cell surface, while the remainder are released into the extracellular environment. The anchored surface proteins play a variety of important roles in cellular processes, ranging from facilitating interactions between cells to maintaining cell stability. The extensively studied S-layer glycoprotein (SLG) of , previously thought to be anchored via C-terminal intercalation into the membrane, was recently shown to be lipidated and to have its C-terminal segment removed in processes dependent upon archaeosortase A (ArtA), a recently discovered enzyme. While SLG is a Sec substrate, analyses presented here reveal that, of eight additional ArtA substrates predicted, two substrates also contain predicted Tat signal peptides, including Hvo_0405, which has a highly conserved tripartite structure that lies closer to the center of the protein than to its C terminus, unlike other predicted ArtA substrates identified to date. We demonstrate that, even given its atypical location, this tripartite structure, which likely resulted from the fusion of genes encoding an ArtA substrate and a cytoplasmic protein, is processed in an ArtA-dependent manner. Using an Hvo_0405 mutant lacking the conserved "twin" arginines of the predicted Tat signal peptide, we show that Hvo_0405 is indeed a Tat substrate and that ArtA substrates include both Sec and Tat substrates. Finally, we confirmed the Tat-dependent localization and signal peptidase I (SPase I) cleavage site of Hvo_0405 using mass spectrometry. The specific mechanisms that facilitate protein anchoring to the archaeal cell surface remain poorly understood. Here, we have shown that the proteins bound to the cell surface of the model archaeon , through a recently discovered novel ArtA-dependent anchoring mechanism, are more structurally diverse than was previously known. Specifically, our results demonstrate that both Tat and Sec substrates, which contain the conserved tripartite structure of predicted ArtA substrates, can be processed in an ArtA-dependent manner and that the tripartite structure need not lie near the C terminus for this processing to occur. These data improve our understanding of archaeal cell biology and are invaluable for subcellular localization predictions of archaeal and bacterial proteins.

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“…This is the usual approach in B. subtilis, where most translocated proteins are secreted into the medium (155,156). In E. coli, export to the periplasm decreases proteolysis and eases downstream purification as the periplasm only contains 4% of the proteome 157,158). Here, proteins are released by osmotic shock (159).…”

Section: Future Perspectives For Biotechnologymentioning

confidence: 99%