Archaeosortases and Exosortases Are Widely Distributed Systems Linking Membrane Transit with Posttranslational Modification

Haft, Daniel H.; Payne, Samuel H.; Selengut, Jeremy D.

doi:10.1128/jb.06026-11

Cited by 67 publications

(118 citation statements)

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“…1A). In silico analysis revealed that the glycine and phenylalanine of this highly conserved motif, found at the start of the conserved C-terminal tripartite structure of predicted ArtA substrates, are invariant, suggesting that the motif is crucial for ArtA recognition (12). Using homologous recombination, the slg G796F,F797G strain was constructed, in which a chromosomal copy of slg was replaced with an slg G796F,F797G mutation and selected for by a trpA gene inserted downstream (see Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Cells lacking ArtA exhibit a growth defect, are relatively small, and have a severe motility defect (14). While some of these phenotypes are consistent with defects that might be caused by an unstable cell wall due to the improper surface anchoring of the unprocessed SLG, H. volcanii is predicted to contain eight additional ArtA substrates, so the failure to accurately process these other substrates might also contribute to the observed phenotypes (12).…”

mentioning

confidence: 98%

“…In silico analyses by Haft et al identified several species of Gram-negative bacteria and archaea that encode a set of Sec substrates that share a C-terminal domain structurally similar to the tripartite structure of the sortase substrates (12,13). While species encoding these putative substrates lack sortase homologs, genome comparisons revealed that each genome encoding such substrates also encodes at least one member of a well-defined protein family, the exosortases and archaeosortases in bacteria and archaea, respectively (12,13).…”

mentioning

confidence: 99%

“…While species encoding these putative substrates lack sortase homologs, genome comparisons revealed that each genome encoding such substrates also encodes at least one member of a well-defined protein family, the exosortases and archaeosortases in bacteria and archaea, respectively (12,13).…”

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confidence: 99%

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Permuting the PGF Signature Motif Blocks both Archaeosortase-Dependent C-Terminal Cleavage and Prenyl Lipid Attachment for the Haloferax volcanii S-Layer Glycoprotein

et al. 2016

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For years, the S-layer glycoprotein (SLG), the sole component of many archaeal cell walls, was thought to be anchored to the cell surface by a C-terminal transmembrane segment. Recently, however, we demonstrated that the Haloferax volcanii SLG C terminus is removed by an archaeosortase (ArtA), a novel peptidase. SLG, which was previously shown to be lipid modified, contains a C-terminal tripartite structure, including a highly conserved proline-glycine-phenylalanine (PGF) motif. Here, we demonstrate that ArtA does not process an SLG variant where the PGF motif is replaced with a PFG motif (slg G796F,F797G ). Furthermore, using radiolabeling, we show that SLG lipid modification requires the PGF motif and is ArtA dependent, lending confirmation to the use of a novel C-terminal lipid-mediated protein-anchoring mechanism by prokaryotes. Similar to the case for the ⌬artA strain, the growth, cellular morphology, and cell wall of the slg G796F,F797G strain, in which modifications of additional H. volcanii ArtA substrates should not be altered, are adversely affected, demonstrating the importance of these posttranslational SLG modifications. Our data suggest that ArtA is either directly or indirectly involved in a novel proteolysis-coupled, covalent lipid-mediated anchoring mechanism. Given that archaeosortase homologs are encoded by a broad range of prokaryotes, it is likely that this anchoring mechanism is widely conserved. IMPORTANCEProkaryotic proteins bound to cell surfaces through intercalation, covalent attachment, or protein-protein interactions play critical roles in essential cellular processes. Unfortunately, the molecular mechanisms that anchor proteins to archaeal cell surfaces remain poorly characterized. Here, using the archaeon H. volcanii as a model system, we report the first in vivo studies of a novel protein-anchoring pathway involving lipid modification of a peptidase-processed C terminus. Our findings not only yield important insights into poorly understood aspects of archaeal biology but also have important implications for key bacterial species, including those of the human microbiome. Additionally, insights may facilitate industrial applications, given that photosynthetic cyanobacteria encode uncharacterized homologs of this evolutionarily conserved enzyme, or may spur development of unique drug delivery systems.A variety of protein complexes decorate cell surfaces, where they support cell stability and facilitate interactions between the cell and the extracellular environment (1-3). Determining the mechanisms that allow these proteins to remain associated with the cell envelope is critical for developing a comprehensive understanding of the biosynthesis of cell surface structures. Protein anchoring to the cell membrane was likely first accomplished by means of transmembrane (TM) segments (4). However, as cell membranes became densely packed with protein complexes, protein-anchoring mechanisms that allowed more economic use of membrane surface area emerged (5). One such mechanism anchors...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 98%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Permuting the PGF Signature Motif Blocks both Archaeosortase-Dependent C-Terminal Cleavage and Prenyl Lipid Attachment for the Haloferax volcanii S-Layer Glycoprotein

et al. 2016

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“…In Gram-positive bacteria, S-layers are linked to the underlying cell wall components, including peptidoglycan and so-called secondary cell wall polymers (SCWPs) (29)(30)(31). In Gram-negative bacteria, S-layers are anchored in the outer membrane, while in Archaea, S-layers are always found to be anchored in the cytoplasmic membrane (32)(33)(34).…”

mentioning

confidence: 99%

A New Addition to the Cell Plan of Anammox Bacteria: "Candidatus Kuenenia stuttgartiensis" Has a Protein Surface Layer as the Outermost Layer of the Cell

Teeseling¹,

Almeida²,

Klingl³

et al. 2013

Journal of Bacteriology

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bAnammox bacteria perform anaerobic ammonium oxidation (anammox) and have a unique compartmentalized cell consisting of three membrane-bound compartments (from inside outwards): the anammoxosome, riboplasm, and paryphoplasm. The cell envelope of anammox bacteria has been proposed to deviate from typical bacterial cell envelopes by lacking both peptidoglycan and a typical outer membrane. However, the composition of the anammox cell envelope is presently unknown. Here, we investigated the outermost layer of the anammox cell and identified a proteinaceous surface layer (S-layer) (a crystalline array of protein subunits) as the outermost component of the cell envelope of the anammox bacterium "Candidatus Kuenenia stuttgartiensis." This is the first description of an S-layer in the phylum of the Planctomycetes and a new addition to the cell plan of anammox bacteria. This S-layer showed hexagonal symmetry with a unit cell consisting of six protein subunits. The enrichment of the S-layer from the cell led to a 160-kDa candidate protein, Kustd1514, which has no homology to any known protein. This protein is present in a glycosylated form. Antibodies were generated against the glycoprotein and used for immunogold localization. The antiserum localized Kustd1514 to the S-layer and thus verified that this protein forms the "Ca. Kuenenia stuttgartiensis" S-layer.

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Random transposon mutagenesis identifies genes essential for transformation in Methanococcus maripaludis

et al. 2023

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Natural transformation, the process whereby a cell acquires DNA directly from the environment, is an important driver of evolution in microbial populations, yet the mechanism of DNA uptake is only characterized in bacteria. To expand our understanding of natural transformation in archaea, we undertook a genetic approach to identify a catalog of genes necessary for transformation in Methanococcus maripaludis. Using an optimized method to generate random transposon mutants, we screened 6144 mutant strains for defects in natural transformation and identified 25 transformation-associated candidate genes. Among these are genes encoding components of the type IV-like pilus, transcription/translation associated genes, genes encoding putative membrane bound transport proteins, and genes of unknown function. Interestingly, similar genes were identified regardless of whether replicating or integrating plasmids were provided as a substrate for transformation. Using allelic replacement mutagenesis, we confirmed that several genes identified in these screens are essential for transformation. Finally, we identified a homolog of a membrane bound substrate transporter in Methanoculleus thermophilus and verified its importance for transformation using allelic replacement mutagenesis, suggesting a conserved mechanism for DNA transfer in multiple archaea. These data represent an initial characterization of the genes important for transformation which will inform efforts to understand gene flow in natural populations. Additionally, knowledge of the genes necessary for natural transformation may assist in identifying signatures of transformation machinery in archaeal genomes and aid the establishment of new model genetic systems for studying archaea.

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Archaeosortases and Exosortases Are Widely Distributed Systems Linking Membrane Transit with Posttranslational Modification

Cited by 67 publications

References 40 publications

Permuting the PGF Signature Motif Blocks both Archaeosortase-Dependent C-Terminal Cleavage and Prenyl Lipid Attachment for the Haloferax volcanii S-Layer Glycoprotein

Permuting the PGF Signature Motif Blocks both Archaeosortase-Dependent C-Terminal Cleavage and Prenyl Lipid Attachment for the Haloferax volcanii S-Layer Glycoprotein

A New Addition to the Cell Plan of Anammox Bacteria: "Candidatus Kuenenia stuttgartiensis" Has a Protein Surface Layer as the Outermost Layer of the Cell

Random transposon mutagenesis identifies genes essential for transformation in Methanococcus maripaludis

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