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Title: Haloferax volcanii archaeosortase is required for motility, mating, and C-terminal processing of the S-layer glycoprotein: Haloferax volcanii archeosortase

Abstract

Cell surfaces are decorated by a variety of proteins that facilitate interactions with their environments and support cell stability.These secreted proteins are anchored to the cell by mechanisms that are diverse, and, in archaea, poorly understood. Recently published in silico data suggest that in some species a subset of secreted euryarchaeal proteins, which includes the S-­layer glycoprotein, is processed and covalently linked tot he cell membrane by enzymes referred to as archaeosortases. In silico work led to the proposal that an independent, sortase-like system for proteolysis-coupled carboxy-terminal lipid modification exists in bacteria (exosortase) and archaea (archaeosortase). Here, we provide the first in vivo characterization of an archaeosortase in the haloarchaeal model organism Haloferax volcanii. Deletion of the artA gene (HVO_0915) resulted in multiple biological phenotypes: (a) poor growth, especially under low-salt conditions, (b) alterations in cell shape and the S-layer, (c) impaired motility, suppressors of which still exhibit poor growth, and (d) impaired conjugation. We studied one of the ArtA substrates, the S-layer glycoprotein, using detailed proteomic analysis. While the carboxy-terminal region of S-layer glycoproteins, consisting of a threonine-rich O-glycosylated region followed by a hydrophobic transmembrane helix, has been notoriously resistant to any proteomic peptide identification, we were able tomore » identify two overlapping peptides from the transmembrane domain present in the ΔartA strain but not in the wild-type strain. This clearly shows that ArtA is involved in carboxy-terminal posttranslational processing of the S-layer glycoprotein. As it is known from previous studies that a lipid is covalently attached to the carboxy-terminal region of the S-layer glycoprotein, our data strongly support the conclusion that archaeosortase functions analogously to sortase, mediating proteolysis-coupled, covalent cell surface attachment.« less

Authors:
 [1];  [2];  [1];  [1];  [3];  [4];  [4];  [4];  [5];  [4];  [1]
  1. University of Pennsylvania, Department of Biology, Philadelphia, PA, 19104, USA
  2. Department of Membrane Biochemistry, Max-Planck-Institute of Biochemistry, 82152, Martinsried, Germany
  3. J. Craig Venter Institute, Rockville, MD, 20850, USA
  4. Environmental Molecular Sciences Laboratory, Richland, WA, USA
  5. Division of Biological Sciences, Pacific Northwest National Laboratory, Richland, WA, USA
Publication Date:
Research Org.:
Pacific Northwest National Laboratory (PNNL), Richland, WA (US), Environmental Molecular Sciences Laboratory (EMSL)
Sponsoring Org.:
USDOE
OSTI Identifier:
1088612
Report Number(s):
PNNL-SA-93808
Journal ID: ISSN 0950-382X; 47650; KP1704020
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Journal Name:
Molecular microbiology (Print)
Additional Journal Information:
Journal Volume: 88; Journal Issue: 6; Journal ID: ISSN 0950-382X
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Environmental Molecular Sciences Laboratory

Citation Formats

Abdul Halim, Mohd Farid, Pfeiffer, Friedhelm, Zou, James, Frisch, Andrew, Haft, Daniel, Wu, Si, Tolić, Nikola, Brewer, Heather, Payne, Samuel H., Paša-Tolić, Ljiljana, and Pohlschroder, Mechthild. Haloferax volcanii archaeosortase is required for motility, mating, and C-terminal processing of the S-layer glycoprotein: Haloferax volcanii archeosortase. United States: N. p., 2013. Web. doi:10.1111/mmi.12248.
Abdul Halim, Mohd Farid, Pfeiffer, Friedhelm, Zou, James, Frisch, Andrew, Haft, Daniel, Wu, Si, Tolić, Nikola, Brewer, Heather, Payne, Samuel H., Paša-Tolić, Ljiljana, & Pohlschroder, Mechthild. Haloferax volcanii archaeosortase is required for motility, mating, and C-terminal processing of the S-layer glycoprotein: Haloferax volcanii archeosortase. United States. doi:10.1111/mmi.12248.
Abdul Halim, Mohd Farid, Pfeiffer, Friedhelm, Zou, James, Frisch, Andrew, Haft, Daniel, Wu, Si, Tolić, Nikola, Brewer, Heather, Payne, Samuel H., Paša-Tolić, Ljiljana, and Pohlschroder, Mechthild. Tue . "Haloferax volcanii archaeosortase is required for motility, mating, and C-terminal processing of the S-layer glycoprotein: Haloferax volcanii archeosortase". United States. doi:10.1111/mmi.12248.
@article{osti_1088612,
title = {Haloferax volcanii archaeosortase is required for motility, mating, and C-terminal processing of the S-layer glycoprotein: Haloferax volcanii archeosortase},
author = {Abdul Halim, Mohd Farid and Pfeiffer, Friedhelm and Zou, James and Frisch, Andrew and Haft, Daniel and Wu, Si and Tolić, Nikola and Brewer, Heather and Payne, Samuel H. and Paša-Tolić, Ljiljana and Pohlschroder, Mechthild},
abstractNote = {Cell surfaces are decorated by a variety of proteins that facilitate interactions with their environments and support cell stability.These secreted proteins are anchored to the cell by mechanisms that are diverse, and, in archaea, poorly understood. Recently published in silico data suggest that in some species a subset of secreted euryarchaeal proteins, which includes the S-­layer glycoprotein, is processed and covalently linked tot he cell membrane by enzymes referred to as archaeosortases. In silico work led to the proposal that an independent, sortase-like system for proteolysis-coupled carboxy-terminal lipid modification exists in bacteria (exosortase) and archaea (archaeosortase). Here, we provide the first in vivo characterization of an archaeosortase in the haloarchaeal model organism Haloferax volcanii. Deletion of the artA gene (HVO_0915) resulted in multiple biological phenotypes: (a) poor growth, especially under low-salt conditions, (b) alterations in cell shape and the S-layer, (c) impaired motility, suppressors of which still exhibit poor growth, and (d) impaired conjugation. We studied one of the ArtA substrates, the S-layer glycoprotein, using detailed proteomic analysis. While the carboxy-terminal region of S-layer glycoproteins, consisting of a threonine-rich O-glycosylated region followed by a hydrophobic transmembrane helix, has been notoriously resistant to any proteomic peptide identification, we were able to identify two overlapping peptides from the transmembrane domain present in the ΔartA strain but not in the wild-type strain. This clearly shows that ArtA is involved in carboxy-terminal posttranslational processing of the S-layer glycoprotein. As it is known from previous studies that a lipid is covalently attached to the carboxy-terminal region of the S-layer glycoprotein, our data strongly support the conclusion that archaeosortase functions analogously to sortase, mediating proteolysis-coupled, covalent cell surface attachment.},
doi = {10.1111/mmi.12248},
journal = {Molecular microbiology (Print)},
issn = {0950-382X},
number = 6,
volume = 88,
place = {United States},
year = {2013},
month = {5}
}